Specificity of bispecific T cell receptors and antibodies targeting peptide-HLA

Tumor-associated peptide–human leukocyte antigen complexes (pHLAs) represent the largest pool of cell surface–expressed cancer-specific epitopes, making them attractive targets for cancer therapies. Soluble bispecific molecules that incorporate an anti-CD3 effector function are being developed to redirect T cells against these targets using 2 different approaches. The first achieves pHLA recognition via affinity-enhanced versions of natural TCRs (e.g., immune-mobilizing monoclonal T cell receptors against cancer [ImmTAC] molecules), whereas the second harnesses an antibody-based format (TCR-mimic antibodies). For both classes of reagent, target specificity is vital, considering the vast universe of potential pHLA molecules that can be presented on healthy cells. Here, we made use of structural, biochemical, and computational approaches to investigate the molecular rules underpinning the reactivity patterns of pHLA-targeting bispecifics. We demonstrate that affinity-enhanced TCRs engage pHLA using a comparatively broad and balanced energetic footprint, with interactions distributed over several HLA and peptide side chains. As ImmTAC molecules, these TCRs also retained a greater degree of pHLA selectivity, with less off-target activity in cellular assays. Conversely, TCR-mimic antibodies tended to exhibit binding modes focused more toward hot spots on the HLA surface and exhibited a greater degree of crossreactivity. Our findings extend our understanding of the basic principles that underpin pHLA selectivity and exemplify a number of molecular approaches that can be used to probe the specificity of pHLA-targeting molecules, aiding the development of future reagents

Redesigning current instruments as a precondition for a more effective environmental protection

The problems of environment and evaluation of natural capital belong to the phenomenology of externalities, and the requirements of environmental economics are ever more oriented towards the elaboration of the ecological-economic category of the externalization of internal expenses, wherefrom reasons for the introduction of ecological-economic instruments, before all payments for pollutions, ensue. Demystification of the negative conviction relating ecological payments as taxes is very important, since it must be shown that ecological payments correct, mitigate or remove externalities. Environmental protection is a complex system and it consists in a set of rules, measures and instruments that are applied for the surveillance of pollution sources, and shaping of instruments for environmental protection is a complex process intended for the accomplishment of established ecological goals. Among all instruments available for bridging the gap of internalization and exerting a significant effect on polluters to diminish their emissions of pollution, the ecological and economic ones are most important. The results of our investigation have demonstrated that it is necessary to reshape the current set of instruments regarding protection of vital functions of nature and an efficient environmental protection, and one of the key cognitions is that the scientific and professional public must not be excluded from the appraisals of creating and redesigning the instruments of environmental protection. These assertions have their foundation in the premise that the significance of elements of the social dimension in sustainable development is to a great extent determined by the level of environmental degradation and reduction in capacity of renewable sources. The paper, approach and data processing of the conducted research offer a new model and new elements of methodology for establishing the state and quality of the environment, whereby social-economic criteria of research (a socio-economic laboratory) are introduced into the fundamental analysis of the condition of crucial media of the environment, but also into shaping a more efficacious instrument set of protection. In such a way, a substantial contribution is given to the affirmation of a holistic and inter-disciplinary approach in the investigation and understanding the phenomenon of contemporary environmental reality, especially the central media of the environment (water, air, soil)

Colletotrichum orbiculare on watermelon: Identification and in vitro inhibition by antagonistic fungi

Anthracnose caused by the fungus Colletotrichum orbiculare is one of the most significant diseases of Cucurbitaceae. In Serbia watermelon fruits with typical anthracnose lesions were collected during the year of 2015. Affected fruits showed sunken, dark brown to black lesions with orange conidial masses produced in black acervuli. In an attempt to identify the causal organism, small pieces of necrotic tissue were surface sterilized and placed on potato dextrose agar (PDA). Macroscopic and microscopic morphologicaly characteristics of three isolates were observed after growth on PDA for 7 days at 25 ºC under a 12 h light/dark cycle. Fungal colonies developed white, grey to black dense aerial mycelium. Conidia were hyaline, aseptate, straight and cylindrical to clavate, 9–12.5 μm × 4–5.5 μm. Fungal isolates were also characterized by sequencing of the internal transcribed spacer (ITS) rDNA region using ITS1F/ITS4 primers and β-tubuline 2 gene using T1/Bt2b primers. The nucleotide sequences were deposited in GenBank (ITS Acc. No. KT454386, KT454387 and KT454388; β-tubuline 2 gene Acc. No. KT581236, KT581237 and KT581238). BLAST analysis of ITS and β-tubuline 2 gene sequences showed that our isolates were 100% identical to other C. orbiculare in NCBI GenBank. Pathogenicity test was conducted on symptomless, detached watermelon fruits. All tested isolates caused anthracnose lesions on watermelon fruits after 10 days of incubation. Trichoderma harzianum (DSM 63059) and Gliocladium roseum (DSM 62726) were evaluated in vitro for their antagonistic potential against C. orbiculare. The results of this study identify T. harzianum and G. roseum as promising biological control agents (BCAs) for further testing against anthracnose disease on watermelon fruits

Translation to Serbian, cultural adaptation, reliability testing and validation of the questionnaire estimating the fear of injections

© 2019, Inst. Sci. inf., Univ. Defence in Belgrade. All rights reserved. Background/Aim. The two-part questionnaire called Injection Phobia Scale (IPS)-Anxiety and IPS-Avoidance represents one of the most commonly used questionnaires for assessing the fear of injections. The aim of the present study was to translate and culturally adapt this questionnaire from English into Serbian as well as to assess reliability and validity of the translation. Methods. The translation and cultural adaptation of the IPS-Anxiety and IPS-Avoidance was performed in accordance with the International Society for Pharmacoeconomics and Outcomes Research (ISPOR) guidelines. Reliability testing, factor analysis and validation of Serbian translation of IPS-Anxiety and IPS-Avoidance were carried out on a sample of 485 students of pharmacy, or medicine at the University of Kragujevac, Serbia. Results. Serbian translation of IPS-Anxiety and IPSAvoidance demonstrated high internal consistency with Cronbach’s alpha of 0.934 for IPS-Anxiety and 0.911 for IPS-Avoidance. Factor analysis of IPS-Anxiety showed that there are two domains, which we have called as Direct Experience (9 items) and Indirect Experience (9 items); factor analysis of IPS-Avoidance also pointed out on two domains referring to direct and indirect fear of injections. Female students scored higher on the scale showing more extensive injection phobia than male students. It is also interesting that students of pharmacy have higher level of injection phobia than students of medicine, and those students of the fifth year of study feel more fear of injections than students from the first four years. Conclusion. Serbian translation of IPS-Anxiety and IPS-Avoidance showed good psychometric properties on population consisted of students medicine and pharmacy

Potential drug-drug interactions in acute ischemic stroke patients at the Neurological Intensive Care Unit

Clinically relevant potential drug-drug interactions are considered preventable adverse drug reactions

The Prospects of Secondary Moderate Mitral Regurgitation after Aortic Valve Replacement —Meta-Analysis

Aortic valve replacement for aortic stenosis represents one of the most frequent surgical procedures on heart valves. These patients often have concomitant mitral regurgitation. To reveal whether the moderate mitral regurgitation will improve after aortic valve replacement alone, we performed a systematic review and meta-analysis. We identified 27 studies with 4452 patients that underwent aortic valve replacement for aortic stenosis and had co-existent mitral regurgitation. Primary end point was the impact of aortic valve replacement on the concomitant mitral regurgitation. Secondary end points were the analysis of the left ventricle reverse remodeling and long-term survival. Our results showed that there was significant improvement in mitral regurgitation postoperatively (RR, 1.65; 95% CI 1.36–2.00; p < 0.00001) with the average decrease of 0.46 (WMD; 95% CI 0.35–0.57; p < 0.00001). The effect is more pronounced in the elderly population. Perioperative mortality was higher (p < 0.0001) and long-term survival significantly worse (p < 0.00001) in patients that had moderate/severe mitral regurgitation preoperatively. We conclude that after aortic valve replacement alone there are fair chances but for only slight improvement in concomitant mitral regurgitation. The secondary moderate mitral regurgitation should be addressed at the time of aortic valve replacement. A more conservative approach should be followed for elderly and high-risk patients

Antigen Potency and Maximal Efficacy Reveal a Mechanism of Efficient T Antigen Potency and Maximal Efficacy Reveal a Mechanism of Efficient T Cell Activation

The following resources related to this article are available online at http://stke.sciencemag.org. Article Tools http://stke.sciencemag.org/cgi/content/full/sigtrans;4/176/ra39 Visit the online version of this article to access the personalization and article tools: T cell activation, a critical event in adaptive immune responses, depends on productive interactions between T cell receptors (TCRs) and antigens presented as peptide-bound major histocompatibility complexes (pMHCs). Activated T cells lyse infected cells, secrete cytokines, and perform other effector functions with various efficiencies, which depend on the binding parameters of the TCR-pMHC complex. The mechanism through which binding parameters are translated to the efficiency of T cell activation, however, remains controversial. The "affinity model" suggests that the dissociation constant (K D ) of the TCR-pMHC complex determines the response, whereas the "productive hit rate model" suggests that the off-rate (k off ) is critical. Here, we used mathematical modeling to show that antigen potency, as determined by the EC 50 (half-maximal effective concentration), which is used to support K D -based models, could not discriminate between the affinity and the productive hit rate models. Both models predicted a correlation between EC 50 and K D , but only the productive hit rate model predicted a correlation between maximal efficacy (E max ), the maximal T cell response induced by pMHC, and k off . We confirmed the predictions made by the productive hit rate model in experiments with cytotoxic T cell clones and a panel of pMHC variants. Thus, we propose that the activity of an antigen is determined by both its potency (EC 50 ) and maximal efficacy (E max ). Material

Doppler Indices of the Uterine, Umbilical and Fetal Middle Cerebral Artery in Diabetic versus Non-Diabetic Pregnancy: Systematic Review and Meta-Analysis

Background and Objectives: The aim of this study was to assess the differences in Doppler indices of the uterine (Ut), umbilical (UA), and middle cerebral artery (MCA) in diabetic versus non-diabetic pregnancies by conducting a comprehensive systematic review of the literature with a meta-analysis. Materials and Methods: PubMed, Web of Science, and SCOPUS were searched for studies that measured the pulsatility index (PI), resistance index (RI), and systolic/diastolic ratio index (S/D ratio) of the umbilical artery, middle cerebral artery, and uterine artery in diabetic versus non-diabetic pregnancies. Two reviewers independently evaluated the eligibility of studies, abstracted data, and performed quality assessments according to standardized protocols. The standardized mean difference (SMD) was used as a measure of effect size. Heterogeneity was assessed using the I2 statistic. Publication bias was evaluated by means of funnel plots. Results: A total of 62 publications were included in the qualitative and 43 in quantitative analysis. The UA-RI, UtA-PI, and UtA-S/D ratios were increased in diabetic compared with non-diabetic pregnancies. Subgroup analysis showed that levels of UtA-PI were significantly higher during the third, but not during the first trimester of pregnancy in diabetic versus non-diabetic pregnancies. No differences were found for the UA-PI, UA-S/D ratio, MCA-PI, MCA-RI, MCA-S/D ratio, or UtA-RI between diabetic and non-diabetic pregnancies. Conclusions: This meta-analysis revealed the presence of hemodynamic changes in uterine and umbilical arteries, but not in the middle cerebral artery in pregnancies complicated by diabetes