bluuebunny/crossref_metadata_2025 huggingface dataset:
A Volumetric Absorptive Microsampling Technique to Monitor Cannabidiol Levels in Epilepsy Patients
Authors: Dubois Sara , Marchese Francesca , Pigliasco Federica , Barco Sebastiano , Tripodi Gino , Lomonaco Tommaso , Lattanzi Simona , Russo Emilio , Cangemi Giuliana , Striano Pasquale
Published: December 2020

Purpose: Interest in cannabis-based therapies has recently increased, due to the availability of cannabidiol (CBD) for the treatment of epilepsy without psychoactive effects. Therapeutic drug monitoring can prevent drug interactions and minimize drug toxicity. We evaluated a volumetric absorptive microsampling (VAMS) method combined with LC-MS/MS (liquid chromatography coupled to tandem mass spectrometry) for the quantification of CBD blood levels in patients with refractory epilepsy.

Methods: Prospective observation of patients with Dravet syndrome receiving open-label, add-on GW-purified CBD (Epidyolex®) at different doses. CBD plasma samples were obtained from venipuncture and LC-MS/MS was used to measure CBD in venous and capillary blood samples collected by VAMS.

Results: We enrolled five patients with a mean age of 13 (range: 4–27) years. CBD levels measured by VAMS on capillary blood did not differ from CBD levels measured in plasma by venipuncture (R2 > 0.93).

Conclusion: This proof-of-concept study suggests that VAMS allows monitoring of CBD plasma levels and can offer valuable support for personalized therapy in refractory epilepsy.
Population Pharmacokinetics and Model-Based Dosing Optimization of Teicoplanin in Pediatric Patients
Authors: Zhang Tao , Sun Dan , Shu Zuocheng , Duan Ziyun , Liu Yang , Du Qian , Zhang Ying , Dong Yuzhu , Wang Taotao , Hu Sasa , Cheng Hua , Dong Yalin
Published: December 2020

Objectives: The pharmacokinetics (PK) of teicoplanin differs in children compared with adults. Our aim was to determine the PK of teicoplanin in an Asian pediatric population and to optimize dosage regimens.

Methods: This was a retrospective PK study and all the data were collected from hospitalized children. We developed a population PK model using sparse data, and Monte Carlo simulation was used to assess the ability of standard teicoplanin regimen and other different dosage regimens. The optimal dosing regimens were defined as achieving the target trough concentration (Cmin) of 10 mg/L and pharmacokinetic/pharmacodynamic (PK/PD, [AUC24/MIC]) of 125 for moderate infection. For severe infection, the optimal dosing regimens were defined as achieving the target 15 mg/L and AUC24/MIC of 345.

Results: 159 children were included and 1.5 samples/children on average were provided. Estimated clearance of teicoplanin was 0.694 L/h (0.784/L/h/70 kg) and volume of distribution was 1.39 L. Teicoplanin standard loading dose was adequate for moderate infection, while 13 mg/kg was needed for severer infection. With standard maintenance doses, both patients with moderate and severe infection failed to achieve the target Cmin. 12 and 16 mg/kg/day were required to achieve a Cmin ≥ 10 and 15 mg/L, respectively. However, standard maintenance dose was adequate to achieve AUC24/MIC ≥ 125 for moderate infection, and 12 mg/kg/day was needed to achieve AUC24/MIC ≥ 345 for severe infection. Lower weight and serum creatinine were associated with higher dose.

Conclusion: Optimal doses based on the target Cmin were higher than that based on the PK/PD target. To achieve the Cmin and PK/PD targets simultaneously, a standard loading dose was adequate for moderate infection based on simulation, while dosing higher than standard doses were required in other situation. Further clinical studies with rich sampling from children is required to confirm our findings.
Cardiac Autonomic Dysfunction and Incidence of de novo Atrial Fibrillation: Heart Rate Variability vs. Heart Rate Complexity
Authors: Wessel Niels , Berg Karsten , Kraemer Jan F. , Gapelyuk Andrej , Rietsch Katrin , Hauser Tino , Kurths Jürgen , Wenzel Dave , Klein Norbert , Kolb Christof , Belke Roberto , Schirdewan Alexander , Kääb Stefan
Published: December 2020

Background

The REACT DX registry evaluates standard therapies to episodes of long-lasting atrial tachyarrhythmias and assesses the quality of sensing and stability of the lead and the implantable cardioverter-defibrillator (ICD) (BIOTRONIK Lumax VR-T DX and successors) over at least a 1-year follow-up period.

Objective

To study the association between the risk of de novo device-detected atrial fibrillation (AF), the autonomic perturbations before the onset of paroxysmal AF and a 7-days heart rate variability (7dHRV) 1 month after ICD implantation.

Methods

The registry consists of 234 patients implanted with an ICD, including 10 with de novo long-lasting atrial tachyarrhythmias with no prior history of AF. The patients were matched via the propensity-score methodology as well as for properties directly influencing the ECGs recorded using GE CardioMem CM 3000. Heart rate variability (HRV) analysis was performed using standard parameters from time- and frequency-domains, and from non-linear dynamics.

Results

No linear HRV was associated with an increased risk of AF (p = n.s.). The only significant approach was derived from symbolic dynamics with the parameter “forbidden words” which distinguished both groups on all 7 days of measurements (p < 0.05), thereby quantifying the heart rate complexity (HRC) as drastically lower in the de novo AF group.

Conclusion

Cardiac autonomic dysfunction denoted by low HRC may be associated with higher AF incidence. For patients with mild to moderate heart failure, standard HRV parameters are not appropriate to quantify cardiac autonomic perturbations before the onset of AF. Further studies are needed to determine the individual risk for AF that would enable interventions to restore autonomic balance in the general population.
Sorting Sox: Diverse Roles for Sox Transcription Factors During Neural Crest and Craniofacial Development
Authors: Schock Elizabeth N. , LaBonne Carole
Published: December 2020

Sox transcription factors play many diverse roles during development, including regulating stem cell states, directing differentiation, and influencing the local chromatin landscape. Of the twenty vertebrate Sox factors, several play critical roles in the development the neural crest, a key vertebrate innovation, and the subsequent formation of neural crest-derived structures, including the craniofacial complex. Herein, we review the specific roles for individual Sox factors during neural crest cell formation and discuss how some factors may have been essential for the evolution of the neural crest. Additionally, we describe how Sox factors direct neural crest cell differentiation into diverse lineages such as melanocytes, glia, and cartilage and detail their involvement in the development of specific craniofacial structures. Finally, we highlight several SOXopathies associated with craniofacial phenotypes.
Expression, Purification and Initial Characterization of Functional α1-Microglobulin (A1M) in Nicotiana benthamiana
Authors: Carlsson Magnus L. R. , Kristiansson Amanda , Bergwik Jesper , Kanagarajan Selvaraju , Bülow Leif , Åkerström Bo , Zhu Li-Hua
Published: December 2020

α1-Microglobulin (A1M) is a small glycoprotein that belongs to the lipocalin protein family. A major biological role of A1M is to protect cells and tissues against oxidative damage by clearing free heme and reactive oxygen species. Because of this, the protein has attracted great interest as a potential pharmaceutical candidate for treatment of acute kidney injury and preeclampsia. The aim of this study was to explore the possibility of expressing human A1M in plants through transient gene expression, as an alternative or complement to other expression systems.E. coli, insect and mammalian cell culture have previously been used for recombinant A1M (rA1M) or A1M production, but these systems have various drawbacks, including additional complication and expense in refolding forE. coli, while insect produced rA1M is heavily modified with chromophores and mammalian cell culture has been used only in analytical scale. For that purpose, we have used a viral vector (pJL-TRBO) delivered byAgrobacteriumfor expression of three modified A1M gene variants in the leaves ofN. benthamiana. The results showed that these modified rA1M protein variants, A1M-NB1, A1M-NB2 and A1M-NB3, targeted to the cytosol, ER and extracellular space, respectively, were successfully expressed in the leaves, which was confirmed by SDS-PAGE and Western blot analysis. The cytosol accumulated A1M-NB1 was selected for further analysis, as it appeared to have a higher yield than the other variants, and was purified with a yield of ca. 50 mg/kg leaf. The purified protein had the expected structural and functional properties, displaying heme-binding capacity and capacity of protecting red blood cells against stress-induced cell death. The protein also carried bound chromophores, a characteristic feature of A1M and an indicator of a capacity to bind small molecules. The study showed that expression of the functional protein inN. benthamianamay be an attractive alternative for production of rA1M for pharmaceutical purposes and a basis for future research on A1M structure and function.
Seed Functional Traits Provide Support for Ecological Restoration and ex situ Conservation in the Threatened Amazon Ironstone Outcrop Flora
Authors: Zanetti Marcilio , Dayrell Roberta L. C. , Wardil Mariana V. , Damasceno Alexandre , Fernandes Tais , Castilho Alexandre , Santos Fernando M. G. , Silveira Fernando A. O.
Published: December 2020

Cangas (ironstone outcrops) host a specialized flora, characterized by high degree of edaphic endemism and an apparent lack of natural history knowledge of its flora. Due to intense pressure from iron ore mining this ecosystem is under threat and in need of restoration. We studied seed functional traits that are relevant for restoration, translocation andex situconservation in 48 species from cangas in eastern Amazon. Were determined the thermal niche breadth, classified seed dormancy and determined methods to overcome it, determined the effect of seed storage on germination, tested the association between germination traits and functional groups, and tested whether seed traits are phylogenetically conserved. We found a broad interspecific variation in most seed traits, except for seed water content. Large interspecific variation in the temperature niche breadth was found among the studied species, but only four species, showed optimum germination at high temperatures of 35–40°C, despite high temperatures under natural conditions. Only 35% of the studied species produced dormant seeds. Mechanical scarification was effective in overcoming physical dormancy and application of gibberellic acid was effective in overcoming physiological dormancy in five species. For the 29 species that seeds were stored for 24 months, 76% showed decreases in the germination percentage. The weak association between germination traits and life-history traits indicate that no particular plant functional type requires specific methods for seed-based translocations. Exceptions were the lianas which showed relatively larger seeds compared to the other growth-forms. Dormancy was the only trait strongly related to phylogeny, suggesting that phylogenetic relatedness may not be a good predictor of regeneration from seeds in cangas. Our study provides support to better manage seed sourcing, use, storage and enhancement techniques with expected reduced costs and increased seedling establishment success.
Hazelnut Pollen Phenotyping Using Label-Free Impedance Flow Cytometry
Authors: Ascari Lorenzo , Cristofori Valerio , Macrì Federico , Botta Roberto , Silvestri Cristian , De Gregorio Tommaso , Huerta Eloy Suarez , Di Berardino Marco , Kaufmann Silvan , Siniscalco Consolata
Published: December 2020

Impedance flow cytometry (IFC) is a versatile lab-on-chip technology which enables fast and label-free analysis of pollen grains in various plant species, promising new research possibilities in agriculture and plant breeding. Hazelnut is a monoecious, anemophilous species, exhibiting sporophytic self-incompatibility. Its pollen is dispersed by wind in midwinter when temperatures are still low and relative humidity is usually high. Previous research found that hazelnut can be characterized by high degrees of pollen sterility following a reciprocal chromosome translocation occurring in some cultivated genotypes. In this study, IFC was used for the first time to characterize hazelnut pollen biology. IFC was validated via dye exclusion in microscopy and employed to (i) follow pollen hydration over time to define the best pre-hydration treatment for pollen viability evaluation; (ii) test hazelnut pollen viability and sterility on 33 cultivars grown in a collection field located in central Italy, and two wild hazelnuts. The accessions were also characterized by their amount and distribution of catkins in the tree canopy. Pollen sterility rate greatly varied among hazelnut accessions, with one main group of highly sterile cultivars and a second group, comprising wild genotypes and the remaining cultivars, producing good quality pollen. The results support the hypothesis of recurring reciprocal translocation events inCorylus avellanacultivars, leading to the observed gametic semi-sterility. The measured hazelnut pollen viability was also strongly influenced by pollen hydration (Radj2= 0.83,P≤ 0.0001) and reached its maximum at around 6 h of pre-hydration in humid chambers. Viable and dead pollen were best discriminated at around the same time of pollen pre-hydration, suggesting that high humidity levels are required for hazelnut pollen to maintain its functionality. Altogether, our results detail the value of impedance flow cytometry for high throughput phenotyping of hazelnut pollen. Further research is required to clarify the causes of pollen sterility in hazelnut, to confirm the role of reciprocal chromosome translocations and to investigate its effects on plant productivity.
Social Networking Sites Addiction and Materialism Among Chinese Adolescents: A Moderated Mediation Model Involving Depression and Need to Belong
Authors: Wang Pengcheng , Lei Li , Yu Guoliang , Li Biao
Published: December 2020

Recent research indicates that social networking site (SNS) addiction is positively associated with materialism. However, little attention has been paid to the potential mechanisms in this relationship. This study tested the mediating role of depression and the moderating role of need to belong (NTB) in the relationships between SNS addiction and adolescents’ materialism. This research model was tested among 733 adolescents in China (mean age = 16.79 years, SD = 0.91). The findings indicated that both SNS addiction and NTB were positively related to adolescents’ materialism. Mediation analyses showed that depression mediated the association between SNS addiction and adolescents’ materialism. Moderated mediation indicated that the effect of SNS addiction on materialism and the effect of SNS addiction on depression were exacerbated by NTB. This study can advance our understanding of how SNS could contribute to adolescents’ materialism in this digital society.
Proinflammatory Cytokines Predict Brain Metabolite Concentrations in the Anterior Cingulate Cortex of Patients With Bipolar Disorder
Authors: Poletti Sara , Mazza Mario Gennaro , Vai Benedetta , Lorenzi Cristina , Colombo Cristina , Benedetti Francesco
Published: December 2020

Bipolar disorder (BD) is a severe psychiatric illness characterized by abnormalities in the immune/inflammatory function and in brain metabolism. Evidences suggest that inflammation may affect the levels of brain metabolites as measured by single-proton magnetic resonance spectroscopy (1H-MRS). The aim of the study was to investigate whether a wide panel of inflammatory markers (i.e., cytokines, chemokines, and growth factors) can predict brain metabolite concentrations of glutamate,myo-inositol,N-acetylaspartate, and glutathione in a sample of 63 bipolar patients and 49 healthy controls. Three cytokines influenced brain metabolite concentrations: IL-9 positively predicts glutamate, IL-1β positively predictsMyo-inositol, and CCL5 positively predictsN-acetylaspartate concentrations. Furthermore, patients showed higher concentrations of glutamate,Myo-inositol, and glutathione and lower concentrations ofN-acetylaspartate in respect to healthy controls. Our results confirm that inflammation in BD alters brain metabolism, through mechanisms possibly including the production of reactive oxygen species and glia activation.
Multilevel Engagements of Pharmacists During the COVID-19 Pandemic: The Way Forward
Authors: Mallhi Tauqeer Hussain , Liaqat Aroosa , Abid Arooj , Khan Yusra Habib , Alotaibi Nasser Hadal , Alzarea Abdulaziz Ibrahim , Tanveer Nida , Khan Tahir Mehmood
Published: December 2020

Severe acute respiratory syndrome caused by the novel coronavirus (SARS-CoV-2) was first reported in China in December 2019 which was later declared to be a public health emergency of international concern by the World Health Organization (WHO). This virus proved to be very contagious resulting in life-threatening respiratory intricacies posing overall public health and governance challenges. Amid the coronavirus pandemic and the unprecedented increase in healthcare demands, only inventive and adaptive practice among healthcare professionals is the need of the hour. Pharmacy services are an important mainstay in the public health and have considerable potential to combat the coronavirus disease 2019 (COVID-19) pandemic. Pharmacists working in several localities and health facilities are linked to patients either directly or indirectly. They can act swiftly in public health response such as drafting professional service guidance to pharmacists working in various healthcare facilities, ensuring effective medicine supply system, monitoring and resolving drug shortage issues, establishing and promoting remote pharmacy services, counseling the public on infection prevention basics, educating about proper use of personal protective equipment, discouraging self-medication, participating in clinical trials, small-scale manufacturing of sanitizers and disinfectants, busting the prevailing myths, and conducting drug evaluation and active surveillance. These interventions will help ease unprecedented burden on healthcare facilities during the ongoing pandemic and eventually will add value to patients and the healthcare system. The current manuscript accentuates the potential roles and activities that pharmacists can initiate in various healthcare facilities to help in relieving pressure on the overwhelmed healthcare system. The information and suggestions offered in this review could help in the restructuring of existing pharmacy services by governments, public health bodies, and policy makers in response to the COVID-19 pandemic. Moreover, this manuscript will underscore any unrealized potential among pharmacists working in various sectors including community, hospital, industry, and drug regulatory authorities.
Microstructure of the Surface of the Tongue and Histochemical Study of the Lingual Glands of the Lowland Tapir (Tapirus terrestris Linnaeus, 1758) (Perissodactyla: Tapiridae)
Authors: Goździewska-Harłajczuk Karolina , Hamouzová Pavla , Klećkowska-Nawrot Joanna , Barszcz Karolina , Čížek Petr
Published: December 2020

Although the anatomy of the gastrointestinal tract has been characterized in the lowland tapir (Tapirus terrestris), the exact anatomy of its tongue has not been studied. Samples of the lingual papillae and lingual glands were collected from the tongue of an adult female lowland tapir. The microscopic analysis of the structure of the lingual papillae and the histochemical analysis of the secretion of the lingual glands were analyzed. The tongue of the tapir is divided into the apex, body with a distinct lingual prominence, and the root. Its ventral surface is smooth. The most numerous of the mechanical papillae were the filiform papillae, while numerous conical papillae with a sharp apex or more rounded papillae were present in the root of the tongue. There were also nine vallate papillae and pair of foliate papillae. The foliate papillae contained several folds parted by 12–14 grooves. The mucous secretion produced by the lingual glands was more obvious than the serous secretion. The features of the dorsal surface of the tongue as well as the shape and number of the lingual papillae on the surface of the tongue of the examined female tapir differ compared to Equidae or Rhinocerotidae, the other two representatives of Perissodactyla. However, further study is necessary for the synapomorpy of the tapir’s tongue.
Global Methylation and Protamine Deficiency in Ram Spermatozoa Correlate with Sperm Production and Quality but Are Not Influenced by Melatonin or Season
Authors: Pool Kelsey R. , Rickard Jessica P. , de Graaf Simon P.
Published: December 2020

This study assessed whether the seasonal effects of melatonin that upregulate ram reproductive function alter sperm global methylation or protamine deficiency and whether these parameters corresponded to ram endocrinology, semen production and quality. Ejaculates were assessed from rams that received melatonin implants (n = 9) or no implants (n = 9) during the non-breeding season. Ejaculates (n = 2/ram/week) were collected prior to implantation (week 0), 1, 6 and 12 weeks post implantation and during the following breeding season (week 30). Flow cytometry was used to assess the sperm global methylation and protamine deficiency in each ejaculate, which had known values for sperm concentration, motility, morphology, DNA fragmentation, seminal plasma levels of melatonin, anti-Mullerian hormone and inhibin A. Serum levels of testosterone and melatonin were also evaluated. Though there was no effect of melatonin or season, sperm protamine deficiency was negatively correlated with sperm production and seminal plasma levels of anti-Mullerian hormone and positively correlated with sperm DNA fragmentation and morphology. Global methylation of spermatozoa was positively correlated with sperm DNA fragmentation, morphology and serum testosterone and negatively correlated with sperm motility. These moderate associations with sperm production and quality suggest that sperm protamine deficiency and global methylation are indicative of ram testicular function.
Administration of Protein Hydrolysates from Anchovy (Engraulis Encrasicolus) Waste for Twelve Weeks Decreases Metabolic Dysfunction-Associated Fatty Liver Disease Severity in ApoE–/–Mice
Authors: Abbate Jessica M. , Macrì Francesco , Capparucci Fabiano , Iaria Carmelo , Briguglio Giovanni , Cicero Luca , Salvo Andrea , Arfuso Francesca , Ieni Antonio , Piccione Giuseppe , Lanteri Giovanni
Published: December 2020

Metabolic dysfunction-associated fatty liver disease (MAFLD) includes several diseases, ranging from simple steatosis to steatohepatitis, fibrosis and cirrhosis. Fish-rich diets are considered helpful in the prevention of MAFLD, and the enzymatic hydrolysis of fish waste has been explored as a means of obtaining high-value protein hydrolysates, which have been proven to exert beneficial bioactivities including anti-obesity and hypocholesterol effects. This study aimed to assess the effect of the administration of protein hydrolysates from anchovy waste (APH) for 12 weeks on attenuated high-fat diet-induced MAFLD in apolipoprotein E-knockout mice (ApoE–/–). Thirty ApoE–/– mice were divided into two groups (n = 15/group) and fed a high-fat diet (HFD), with and without the addition of 10% (w/w) APH. After 12 weeks, serum and hepatic lipid profiles, hepatic enzyme activities, liver histology and immunohistochemistry were analyzed to assess hepatic steatosis, inflammation and fibrosis. Twelve-weeks on a 10% (w/w) APH diet reduces total cholesterol and triglyceride serum levels, hepatic enzyme activity and hepatic triacylglycerol content (p < 0.0001), and results in a reduction in hepatic fat accumulation and macrophage recruitment (p < 0.0001). The results suggest that a 10% APH diet has an anti-obesity effect, with an improvement in lipid metabolism, hepatic steatosis and liver injury as a result of a high-fat diet. Protein hydrolysates from fish waste may represent an efficient nutritional strategy in several diseases, and their use as nutraceuticals is worthy of future investigation.
Epithelial to Mesenchymal Transition (EMT) in a Laryngeal Squamous Cell Carcinoma of a Horse: Future Perspectives
Authors: Armando Federico , Godizzi Francesco , Razzuoli Elisabetta , Leonardi Fabio , Angelone Mario , Corradi Attilio , Meloni Daniela , Ferrari Luca , Passeri Benedetta
Published: December 2020

Squamous cell carcinoma (SCC) is one of the most frequent tumors of skin and muco-cutaneous junctions in the horse. Equine papillomavirus type 2 (EcPV2) has been detected in equine SCC of the oral tract and genitals, and recently also in the larynx. As human squamous cell carcinoma of the larynx (SCCL), it is strongly etiologically associated with high-risk papillomavirus (h-HPV) infection. This study focuses on tumor cells behavior in a naturally occurring tumor that can undergo the so-called epithelial to mesenchymal transition (EMT). A SCCL in a horse was investigated by immunohistochemistry using antibodies against E-cadherin, pan-cytokeratin AE3/AE1, β-catenin, N-cadherin, vimentin, ZEB-1, TWIST, and HIF-1α. EcPV2 DNA detection and expression of oncogenes in SCC were investigated. A cadherin switch and an intermediate filaments rearrangement within primary site tumor cells together with the expression of the EMT-related transcription factors TWIST-1, ZEB-1, and HIF-1α were observed. DNA obtained from the tumor showed EcPV2 positivity, with E2 gene disruption and E6 gene dysregulation. The results suggest that equine SCCL might be a valuable model for studying EMT and the potential interactions between EcPV2 oncoproteins and the EMT process in SCCL.
Potential Use of Hyssopus officinalis and Borago officinalis as Curing Ingredients in Pork Meat Formulations
Authors: Zając Marzena , Duda Iwona , Skoczylas Łukasz , Tabaszewska Małgorzata
Published: December 2020

The replacement of nitrites in pork meat products has been a studied issue for many years. Due to potential health threats associated with these additives, consumers tend to search for alternative meat curing methods. In this study, Hyssopus officinalis and Borago officinalis were tested for their potential to be used as colour-forming and antioxidant agents. Dry plant samples from various sources were tested for fat, protein, ash, polyphenol and nitrate content. There were significant differences between the herbs depending on source. Two control samples (containing curing salt and sodium chloride with nitrate reducing bacteria) and samples with herbs (hyssop, hyssop with nitrate reducing bacteria, borage, borage with nitrate reducing bacteria)—0.5% of the meat mass—were prepared and stored for 15 days. In the samples with herbs and bacterial cultures, a red colour was developed, the TBARS values were low and DPPH activity was strong. All the samples with herbs had lower residual nitrite levels compared to the samples with curing salt. Borage had a stronger influence on colour and antioxidant stability of the meat samples compared to hyssop. However, both herbs can be used as colour-forming and antioxidant agents along with nitrate-reducing bacteria.
Human Behaviour Change Interventions in Animal Care and Interactive Settings: A Review and Framework for Design and Evaluation
Authors: Glanville Carmen , Abraham Charles , Coleman Grahame
Published: December 2020

Behaviour change interventions may be one of the most promising avenues to improve animal welfare. Yet there has been limited systematic research involving them in animal-related settings. We searched three major databases for studies involving an intervention to change interactive or care-related behaviours in any animal-related setting. Forty-seven papers were included in the review and each paper was coded for specific design and evaluation elements. We found a series of limitations in the quality and consistency of intervention design, evaluation, and reporting. Hence, we present a framework, the “Ten-Task” guide, based on the intervention mapping framework, to guide future work in this field. Adopting this structured approach will improve the quality and efficacy of behaviour change interventions for animal welfare and allow for the field to progress in a harmonious way.
Parental Education, Household Income, Race, and Children’s Working Memory: Complexity of the Effects
Authors: Akhlaghipour Golnoush , Assari Shervin
Published: December 2020

Background. Considerable research has linked social determinants of health (SDoHs) such as race, parental education, and household income to school performance, and these effects may be in part due to working memory. However, a growing literature shows that these effects may be complex: while the effects of parental education may be diminished for Blacks than Whites, household income may explain such effects. Purpose. Considering race as sociological rather than a biological construct (race as a proxy of racism) and built on Minorities’ Diminished Returns (MDRs), this study explored complexities of the effects of SDoHs on children’s working memory. Methods. We borrowed data from the Adolescent Brain Cognitive Development (ABCD) study. The total sample was 10,418, 9- and 10-year-old children. The independent variables were race, parental education, and household income. The primary outcome was working memory measured by the NIH Toolbox Card Sorting Test. Age, sex, ethnicity, and parental marital status were the covariates. To analyze the data, we used mixed-effect regression models. Results. High parental education and household income were associated with higher and Black race was associated with lower working memory. The association between high parental education but not household income was less pronounced for Black than White children. This differential effect of parental education on working memory was explained by household income. Conclusions. For American children, parental education generates unequal working memory, depending on race. This means parental education loses some of its expected effects for Black families. It also suggests that while White children with highly educated parents have the highest working memory, Black children report lower working memory, regardless of their parental education. This inequality is mainly because of differential income in highly educated White and Black families. This finding has significant public policy and economic implications and suggests we need to do far more than equalizing education to eliminate racial inequalities in children’s cognitive outcomes. While there is a need for multilevel policies that reduce the effect of racism and social stratification for middle-class Black families, equalizing income may have more returns than equalizing education.
A Matching-Adjusted Indirect Comparison of Pembrolizumab + Chemotherapy vs. Nivolumab + Ipilimumab as First-Line Therapies in Patients with PD-L1 TPS ≥1% Metastatic NSCLC
Authors: Halmos Balazs , Burke Thomas , Kalyvas Chrysostomos , Insinga Ralph , Vandormael Kristel , Frederickson Andrew , Piperdi Bilal
Published: December 2020

Background: In the absence of head-to-head trials, this study indirectly compared the effectiveness of pembrolizumab + chemotherapy vs nivolumab + ipilimumab for the first-line treatment of metastatic stage IV NSCLC patients with PD-L1 tumor proportion score (TPS) ≥1%. Methods: An anchored matching-adjusted indirect comparison (MAIC) was conducted using pooled individual patient data (IPD) from the ITT population in KEYNOTE-021G, KEYNOTE-189 and KEYNOTE-407 (n = 816) and published aggregate data of nivolumab + ipilimumab from CheckMate 227 Part 1A (n = 793). To adjust for cross-trial differences in baseline characteristics, data from KEYNOTE-021G/KEYNOTE-189/KEYNOTE-407 were re-weighted to match the baseline characteristics of CheckMate 227 Part 1A. Outcomes included OS, PFS and ORR. Base case analyses were restricted to patients with PD-L1 TPS ≥1%, with sub-group analyses in PD-L1 TPS ≥50% and 1–49%. Results: The estimated HR (95% CI) of pembrolizumab + chemotherapy vs nivolumab + ipilimumab was 0.80 (0.59,1.09) and 0.53 (0.41,0.68) for OS and PFS, respectively. For ORR, the estimated risk ratio was 1.8 (1.3,2.4) for pembrolizumab + chemotherapy vs nivolumab + ipilimumab and the risk difference was 25.5% (15.0,36.0). PD-L1 TPS ≥50% and 1–49% sub-groups showed an OS HR of 0.89 (0.58,1.36) and 0.68 (0.46,1.01), respectively. Conclusion: These MAIC results suggest that pembrolizumab + chemotherapy leads to a greater clinical benefit vs nivolumab + ipilimumab in patients with PD-L1 TPS ≥1% across multiple endpoints.
Proof of Concept Study for Increasing Tenascin-C-Targeted Drug Delivery to Tumors Previously Subjected to Therapy: X-Irradiation Increases Tumor Uptake
Authors: Sugyo Aya , Tsuji Atsushi B. , Sudo Hitomi , Takano Kanako , Kusakabe Moriaki , Higashi Tatsuya
Published: December 2020

In treatment-refractory cancers, tumor tissues damaged by therapy initiate the repair response; therefore, tumor tissues must be exposed to an additional burden before successful repair. We hypothesized that an agent recognizing a molecule that responds to anticancer treatment-induced tissue injury could deliver an additional antitumor agent including a radionuclide to damaged cancer tissues during repair. We selected the extracellular matrix glycoprotein tenascin-C (TNC) as such a molecule, and three antibodies recognizing human and murine TNC were employed to evaluate X-irradiation-induced changes in TNC uptake by subcutaneous tumors. TNC expression was assessed by immunohistochemical staining of BxPC-3 tumors treated with or without X-irradiation (30 Gy) for 7 days. Antibodies against TNC (3–6, 12–2–7, TDEAR) and a control antibody were radiolabeled with 111In and injected into nude mice having BxPC-3 tumors 7 days after X-irradiation, and temporal uptake was monitored for an additional 4 days by biodistribution and single-photon emission computed tomography with computed tomography (SPECT/CT) studies. Intratumoral distribution was analyzed by autoradiography. The immunohistochemical signal for TNC expression was faint in nontreated tumors but increased and expanded with time until day 7 after X-irradiation. Biodistribution studies revealed increased tumor uptake of all three 111In-labeled antibodies and the control antibody. However, a statistically significant increase in uptake was evident only for 111In-labeled 3–6 (35% injected dose (ID)/g for 30 Gy vs. 15% ID/g for 0 Gy at day 1, p < 0.01), whereas limited changes in 111In-labeled TDEAR2, 12–2–27, and control antibody were observed (several % ID/g for 0 and 30 Gy). Serial SPECT/CT imaging with 111In-labeled 3–6 or control antibody provided consistent results. Autoradiography revealed noticeably stronger signals in irradiated tumors injected with 111In-labeled 3–6 compared with each of the nonirradiated tumors and the control antibody. The signals were observed in TNC-expressing stroma. Markedly increased uptake of 111In-labeled 3–6 in irradiated tumors supports our concept that an agent, such as an antibody, that recognizes a molecule involved in tissue injury repair, such as TNC, could enhance drug delivery to tumor tissues that have undergone therapy. The combination of antibody 3–6 coupled to a tumoricidal drug and conventional therapy has the potential to achieve better outcomes for patients with refractory cancer.
Platelet-Derived GARP Induces Peripheral Regulatory T Cells—Potential Impact on T Cell Suppression in Patients with Melanoma-Associated Thrombocytosis
Authors: Zimmer Niklas , Krebs Franziska K. , Zimmer Sophia , Mitzel-Rink Heidrun , Kumm Elena J. , Jurk Kerstin , Grabbe Stephan , Loquai Carmen , Tuettenberg Andrea
Published: December 2020

Platelets have been recently described as an important component of the innate and adaptive immunity through their interaction with immune cells. However, information on the platelet–T cell interaction in immune-mediated diseases remains limited. Glycoprotein A repetitions predominant (GARP) expressed on platelets and on activated regulatory T cells (Treg) is involved in the regulation of peripheral immune responses by modulating the bioavailability of transforming growth factor β (TGF-β). Soluble GARP (sGARP) exhibits strong regulatory and anti-inflammatory capacities both in vitro and in vivo, leading to the induction of peripheral Treg. Herein, we investigated the effect of platelet-derived GARP on the differentiation, phenotype, and function of T effector cells. CD4+CD25− T cells cocultured with platelets upregulated FoxP3, the master transcription factor for Treg, were anergic, and were strongly suppressive. These effects were reversed by using a blocking anti-GARP antibody, indicating a dependency on GARP. Importantly, melanoma patients in different stages of disease showed a significant upregulation of GARP on the platelet surface, correlating to a reduced responsiveness to immunotherapy. In conclusion, our data indicate that platelets induce peripheral Treg via GARP. These findings might contribute to diseases such as cancer-associated thrombocytosis, wherein poor prognosis and metastasis are associated with high counts of circulating platelets.
Prospects for Using Expression Patterns of Paramyxovirus Receptors as Biomarkers for Oncolytic Virotherapy
Authors: Matveeva Olga V. , Shabalina Svetlana A.
Published: December 2020

The effectiveness of oncolytic virotherapy in cancer treatment depends on several factors, including successful virus delivery to the tumor, ability of the virus to enter the target malignant cell, virus replication, and the release of progeny virions from infected cells. The multi-stage process is influenced by the efficiency with which the virus enters host cells via specific receptors. This review describes natural and artificial receptors for two oncolytic paramyxoviruses, nonpathogenic measles, and Sendai viruses. Cell entry receptors are proteins for measles virus (MV) and sialylated glycans (sialylated glycoproteins or glycolipids/gangliosides) for Sendai virus (SeV). Accumulated published data reviewed here show different levels of expression of cell surface receptors for both viruses in different malignancies. Patients whose tumor cells have low or no expression of receptors for a specific oncolytic virus cannot be successfully treated with the virus. Recent published studies have revealed that an expression signature for immune genes is another important factor that determines the vulnerability of tumor cells to viral infection. In the future, a combination of expression signatures of immune and receptor genes could be used to find a set of oncolytic viruses that are more effective for specific malignancies.
Extracellular Cystatin F Is Internalised by Cytotoxic T Lymphocytes and Decreases Their Cytotoxicity
Authors: Prunk Mateja , Perišić Nanut Milica , Jakoš Tanja , Sabotič Jerica , Švajger Urban , Kos Janko
Published: December 2020

Cystatin F is a protein inhibitor of cysteine cathepsins, peptidases involved in the activation of the effector molecules of the perforin/granzyme pathway. Cystatin F was previously shown to regulate natural killer cell cytotoxicity. Here, we show that extracellular cystatin F has a role in regulating the killing efficiency of cytotoxic T lymphocytes (CTLs). Extracellular cystatin F was internalised into TALL-104 cells, a cytotoxic T cell line, and decreased their cathepsin C and H activity. Correspondingly, granzyme A and B activity was also decreased and, most importantly, the killing efficiency of TALL-104 cells as well as primary human CTLs was reduced. The N-terminally truncated form of cystatin F, which can directly inhibit cathepsin C (unlike the full-length form), was more effective than the full-length inhibitor. Furthermore, cystatin F decreased cathepsin L activity, which, however, did not affect perforin processing. Cystatin F derived from K-562 target cells could also decrease the cytotoxicity of TALL-104 cells. These results clearly show that, by inhibiting cysteine cathepsin proteolytic activity, extracellular cystatin F can decrease the cytotoxicity of CTLs and thus compromise their function.
Insights on TAM Formation from a Boolean Model of Macrophage Polarization Based on In Vitro Studies
Authors: Marku Malvina , Verstraete Nina , Raynal Flavien , Madrid-Mencía Miguel , Domagala Marcin , Fournié Jean-Jacques , Ysebaert Loïc , Poupot Mary , Pancaldi Vera
Published: December 2020

The tumour microenvironment is the surrounding of a tumour, including blood vessels, fibroblasts, signaling molecules, the extracellular matrix and immune cells, especially neutrophils and monocyte-derived macrophages. In a tumour setting, macrophages encompass a spectrum between a tumour-suppressive (M1) or tumour-promoting (M2) state. The biology of macrophages found in tumours (Tumour Associated Macrophages) remains unclear, but understanding their impact on tumour progression is highly important. In this paper, we perform a comprehensive analysis of a macrophage polarization network, following two lines of enquiry: (i) we reconstruct the macrophage polarization network based on literature, extending it to include important stimuli in a tumour setting, and (ii) we build a dynamical model able to reproduce macrophage polarization in the presence of different stimuli, including the contact with cancer cells. Our simulations recapitulate the documented macrophage phenotypes and their dependencies on specific receptors and transcription factors, while also unravelling the formation of a special type of tumour associated macrophages in an in vitro model of chronic lymphocytic leukaemia. This model constitutes the first step towards elucidating the cross-talk between immune and cancer cells inside tumours, with the ultimate goal of identifying new therapeutic targets that could control the formation of tumour associated macrophages in patients.
Mi-RNA-888-5p Is Involved in S-Adenosylmethionine Antitumor Effects in Laryngeal Squamous Cancer Cells
Authors: Pagano Martina , Mosca Laura , Vitiello Francesca , Ilisso Concetta Paola , Coppola Alessandra , Borzacchiello Luigi , Mele Luigi , Caruso Francesca Pia , Ceccarelli Michele , Caraglia Michele , Cacciapuoti Giovanna , Porcelli Marina
Published: December 2020

(1) Purpose: The methyl donor S-Adenosylmethionine (AdoMet) has been widely explored as a therapeutic compound, and its application-alone or in combination with other molecules-is emerging as a potential effective strategy for the treatment and chemoprevention of tumours. In this study, we investigated the antitumor activity of AdoMet in Laryngeal Squamous Cell Carcinoma (LSCC), exploring the underlying mechanisms. (2) Results: We demonstrated that AdoMet induced ROS generation and triggered autophagy with a consistent increase in LC3B-II autophagy-marker in JHU-SCC-011 and HNO210 LSCC cells. AdoMet induced ER-stress and activated UPR signaling through the upregulation of the spliced form of XBP1 and CHOP. To gain new insights into the molecular mechanisms underlying the antitumor activity of AdoMet, we evaluated the regulation of miRNA expression profile and we found a downregulation of miR-888-5p. We transfected LSCC cells with miR-888-5p inhibitor and exposed the cells to AdoMet for 48 and 72 h. The combination of AdoMet with miR-888-5p inhibitor synergistically induced both apoptosis and inhibited cell migration paralleled by the up-regulation of MYCBP and CDH1 genes and of their targets. (3) Conclusion: Overall, these data highlighted that epigenetic reprogramming of miRNAs by AdoMet play an important role in inhibiting apoptosis and migration in LSCC cell lines.
NEK1 Phosphorylation of YAP Promotes Its Stabilization and Transcriptional Output
Authors: Khalil Md Imtiaz , Ghosh Ishita , Singh Vibha , Chen Jing , Zhu Haining , De Benedetti Arrigo
Published: December 2020

Most prostate cancer (PCa) deaths result from progressive failure in standard androgen deprivation therapy (ADT), leading to metastatic castration-resistant PCa (mCRPC); however, the mechanism and key players leading to this are not fully understood. While studying the role of tousled-like kinase 1 (TLK1) and never in mitosis gene A (NIMA)-related kinase 1 (NEK1) in a DNA damage response (DDR)-mediated cell cycle arrest in LNCaP cells treated with bicalutamide, we uncovered that overexpression of wt-NEK1 resulted in a rapid conversion to androgen-independent (AI) growth, analogous to what has been observed when YAP1 is overexpressed. We now report that overexpression of wt-NEK1 results in accumulation of YAP1, suggesting the existence of a TLK1>NEK1>YAP1 axis that leads to adaptation to AI growth. Further, YAP1 is co-immunoprecipitated with NEK1. Importantly, NEK1 was able to phosphorylate YAP1 on six residues in vitro, which we believe are important for stabilization of the protein, possibly by increasing its interaction with transcriptional partners. In fact, knockout (KO) of NEK1 in NT1 PCa cells resulted in a parallel decrease of YAP1 level and reduced expression of typical YAP-regulated target genes. In terms of cancer potential implications, the expression of NEK1 and YAP1 proteins was found to be increased and correlated in several cancers. These include PCa stages according to Gleason score, head and neck squamous cell carcinoma, and glioblastoma, suggesting that this co-regulation is imparted by increased YAP1 stability when NEK1 is overexpressed or activated by TLK1, and not through transcriptional co-expression. We propose that the TLK1>NEK1>YAP1 axis is a key determinant for cancer progression, particularly during the process of androgen-sensitive to -independent conversion during progression to mCRPC.
Current and New Biomarkers for Early Detection, Prognostic Stratification, and Management of Gallbladder Cancer Patients
Authors: García Patricia , Lamarca Angela , Díaz Javier , Carrera Enrique , Roa Juan ,   on behalf of the European-Latin American ESCALON Consortium
Published: December 2020

Gallbladder cancer (GBC) is an aggressive disease that shows evident geographic variation and is characterized by a poor prognosis, mainly due to the late diagnosis and ineffective treatment. Genetic variants associated with GBC susceptibility, including polymorphisms within the toll-like receptors TLR2 and TLR4, the cytochrome P450 1A1 (CYP1A1), and the ATP-binding cassette (ABC) transporter ABCG8 genes, represent promising biomarkers for the stratification of patients at higher risk of GBC; thus, showing potential to prioritize cholecystectomy, particularly considering that early diagnosis is difficult due to the absence of specific signs and symptoms. Similarly, our better understanding of the gallbladder carcinogenic processes has led to identify several cellular and molecular events that may influence patient management, including HER2 aberrations, high tumor mutational burden, microsatellite instability, among others. Despite these reports on interesting and promising markers for risk assessment, diagnosis, and prognosis; there is an unmet need for reliable and validated biomarkers that can improve the management of GBC patients and support clinical decision-making. This review article examines the most potentially significant biomarkers of susceptibility, diagnosis, prognosis, and therapy selection for GBC patients, highlighting the need to find and validate existing and new molecular biomarkers to improve patient outcomes.