bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Branch Number on Quality Traits and Yield Properties of European Hazelnut (Corylus avellana L.)
Authors: Bak Tuba , Karadeniz Turan
Published: May 2021

Despite being the world leader in the production of hazelnuts, Turkey’s nut yield per hectare is quite behind other producer countries. The main reasons for low yield are the completion of the economic life of hazelnut orchards and the lack of pruning following the technique. Hazelnuts are being grown with the ‘ocak’ system in Turkey. The ocak system is a bush-like growing technique that does not contain the main trunk but consists of shoots instead. It is necessary to maintain the optimum number of shoots per ocak to increase yield and quality in this system. The effects of different branch numbers on yield and fruit quality parameters of ‘Tombul’ and ‘Palaz’ hazelnut cultivars were investigated in Ordu ecological conditions. ‘Palaz’ had the highest yield per ocak in 8 branches (1460.19 g), while ‘Tombul’ had in the 5 branched ocak (2170.47 g). ‘Palaz’ and ‘Tombul’ had the highest fruit weights in 4 and 4–6 branches, respectively. The highest oil ratio of ‘Palaz’ was observed in 4 branches, while ‘Tombul’ had the highest in 5 branches. The highest kernel ratio was determined in 6 and 5 branches in ‘Palaz’ and ‘Tombul’, respectively. The ocaks with 4 and 5 branches were mostly expressed higher values, and it is foreseen that the number of branches should not exceed 6 to reduce the competition of the branches and to obtain good quality products.
The Microflora of Maize Grains as a Biological Barrier against the Late Wilt Causal Agent, Magnaporthiopsis maydis
Authors: Degani Ofir , Regev Danielle , Dor Shlomit
Published: May 2021

The maize pathogen Magnaporthiopsis maydis causes severe damage to commercial fields in the late growth stages. This late wilt disease has spread since its discovery (the 1980s) and is now common in most corn-growing areas in Israel. In some fields and sensitive plant species, the disease can affect 100% of the plants. The M. maydis pathogen has a hidden endophytic lifecycle (developed inside the plants with no visible symptoms) in resistant corn plants and secondary hosts, such as green foxtail and cotton. As such, it may also be opportunist and attack the host in exceptional cases when conditions encourage it. This work aims to study the pathogen’s interactions with maize endophytes (which may play a part in the plant’s resistance factors). For this purpose, 11 fungal and bacterial endophytes were isolated from six sweet and fodder corn cultivars with varying susceptibility to late wilt disease. Of these, five endophytes (four species of fungi and one species of bacteria) were selected based on their ability to repress the pathogen in a plate confrontation test. The selected isolates were applied in seed inoculation and tested in pots in a growth room with the Prelude maize cultivar (a late wilt-sensitive maize hybrid) infected with the M. maydis pathogen. This assay was accompanied by real-time qPCR that enables tracking the pathogen DNA inside the host roots. After 42 days, two of the endophytes, the Trichoderma asperellum, and Chaetomium subaffine fungi, significantly (p < 0.05) improved the infected plants’ growth indices. The fungal species T. asperellum, Chaetomium cochliodes, Penicillium citrinum, and the bacteria Bacillus subtilis treatments were able to reduce the M. maydis DNA in the host plant’s roots. Studying the maize endophytes’ role in restricting the invasion and devastating impact of M. maydis is an essential initial step towards developing new measures to control the disease. Such an environmentally friendly control interface will be based on strengthening the plants’ microbiome.
Long-Term Preservation and Storage of Faecal Samples in Whatman® Cards for PCR Detection and Genotyping of Giardia duodenalis and Cryptosporidium hominis
Authors: Köster Pamela Carolina , Bailo Begoña , Dashti Alejandro , Hernández-Castro Carolina , Calero-Bernal Rafael , Ponce-Gordo Francisco , González-Barrio David , Carmena David
Published: May 2021

Preservation and conservation of biological specimens, including faecal samples, is a challenge in remote areas or poor-resource settings where the cold chain cannot be maintained. This study aims at evaluating the suitability of filter cards for long-term storage of faecal samples of animal and human origin positive to the diarrhoea-causing protozoan parasites, Giardia duodenalis and Cryptosporidium hominis. Three commercially available Whatman® Filter Cards were comparatively assessed: the FTA® Classic Card, the FTA® Elute Micro Card, and the 903 Protein Saver Card. Human faecal samples positive to G. duodenalis (n = 5) and C. hominis (n = 5) were used to impregnate the selected cards at given storage (1 month, 3 months, and 6 months) periods and temperature (−20 °C, 4 °C, and room temperature) conditions. Parasite DNA was detected by PCR-based methods. Sensitivity assays and quality control procedures to assess suitability for genotyping purposes were conducted. Overall, all three Whatman® cards were proven useful for the detection and molecular characterisation of G. duodenalis and C. hominis under the evaluated conditions. Whatman® cards represent a simple, safe, and cost-effective option for the transportation, preservation, and storage of faecal samples without the need of the cold chain.
Choice of Commercial DNA Extraction Method Does Not Affect 16S Sequencing Outcomes in Cloacal Swabs
Authors: Van Syoc Emily , Carrillo Gaeta Natália , Ganda Erika
Published: May 2021

As the applications of microbiome science in agriculture expand, laboratory methods should be constantly evaluated to ensure optimization and reliability of downstream results. Most animal microbiome research uses fecal samples or rectal swabs for profiling the gut bacterial community; however, in birds, this is difficult given the unique anatomy of the cloaca where the fecal, urinary, and reproductive tracts converge into one orifice. Therefore, avian gut microbiomes are usually sampled from cloacal swabs, creating a need to evaluate sample preparation methods to optimize 16S sequencing. We compared four different DNA extraction methods from two commercially available kits on cloacal swabs from 10 adult commercial laying hens and included mock communities and negative controls, which were then subjected to 16S rRNA amplicon sequencing. Extracted DNA yield and quality, diversity analyses, and contaminants were assessed. Differences in DNA quality and quantity were observed, and all methods needed further purification for optimal sequencing, suggesting contaminants due to cloacal contents, method reagents, and/or environmental factors. However, no differences were observed in alpha or beta diversity between methods. Importantly, multiple bacterial contaminants were detected in each mock community and negative control, indicating the prevalence of laboratory and handling contamination as well as method-specific reagent contamination. We found that although the extraction methods resulted in different extraction quality and yield, overall sequencing results were not affected, and we did not identify any method that would be an inappropriate choice in extracting DNA from cloacal swabs for 16S rRNA sequencing. Overall, our results highlight the need for careful consideration of positive and negative controls in addition to DNA isolation method and lend guidance to future microbiome research in poultry.
The Concentration of Selected Heavy Metals in Muscles, Liver and Kidneys of Pigs Fed Standard Diets and Diets Containing 60% of New Rye Varieties
Authors: Chałabis-Mazurek Agnieszka , Valverde Piedra Jose Luis Valverde , Muszyński Siemowit , Tomaszewska Ewa , Szymańczyk Sylwia , Kowalik Sylwester , Arciszewski Marcin B. , Zacharko-Siembida Anna , Schwarz Tomasz
Published: May 2021

The carry-over of heavy metals from feed to muscles is generally low if animals are fed with a standard diet containing amounts below the maximum permissible levels. However, prolonged exposure to heavy metals can lead to their accumulation in some organs like muscles, liver, and kidneys. This paves the way for human health risks related to the consumption of products of animal-origin. Thus, using feed mixtures with a low level of heavy metals in pig production will contribute to increasing public health and safety and is of environmental concern. The study aimed to assess the impact of the level of some heavy metals (Cd, Pb, Hg, Cu, Zn, Fe and Mn) in standard (control) feed mixtures and in alternative feed mixtures based on maize or new rye varieties (population and hybrid) on the heavy metal concentration in muscles, liver and kidney of fattened pigs at slaughter. While some differences between heavy metals content in examined tissue samples from experimental groups were observed, all of them were in the range of allowable levels according to European Community rules. In conclusion, new rye varieties, especially the hybrid variety, could be an alternative source of cereal grains for pig nutrition.
The Escalating Effects of Wildlife Tourism on Human–Wildlife Conflict
Authors: Cui Qingming , Ren Yuejia , Xu Honggang
Published: May 2021

Human–wildlife conflict is a barrier to achieving sustainable biodiversity conservation and community development in protected areas. Tourism is often regarded as a tool to mitigate such conflict. However, existing studies have mainly adopted a socio-economic perspective to examine the benefits of tourism for communities, neglecting the ecological effects of tourism. This case study of macaque tourism on a peninsula in China illustrates that tourism can escalate rather than mitigate human–wildlife conflict. Fifty-three stakeholders were interviewed and secondary data were collected to understand the development of rhesus macaque (Macaca mulatta) tourism and community–macaque conflict. The results show that food provision and tourist–macaque interactions rapidly increased the macaques’ population, habituation, and aggressive behaviors, which led them to invade the surrounding community more often and exacerbated human–macaque conflict. Meanwhile, low community participation in tourism generated few benefits for residents and did not help alter residents’ hostile attitudes towards the macaques. Local residents gradually retreated from agriculture as the macaques became more intrusive. A holistic approach to evaluating the role of wildlife tourism in resolving community–wildlife conflict is proposed and practical suggestions for alleviating such conflict are given.
Antibiotic-Resistant Enterobacteriaceae in Wastewater of Abattoirs
Authors: Homeier-Bachmann Timo , Heiden Stefan E. , Lübcke Phillip K. , Bachmann Lisa , Bohnert Jürgen A. , Zimmermann Dirk , Schaufler Katharina
Published: May 2021

Antibiotic-resistant Enterobacteriaceae are regularly detected in livestock. As pathogens, they cause difficult-to-treat infections and, as commensals, they may serve as a source of resistance genes for other bacteria. Slaughterhouses produce significant amounts of wastewater containing antimicrobial-resistant bacteria (AMRB), which are released into the environment. We analyzed the wastewater from seven slaughterhouses (pig and poultry) for extended-spectrum β-lactamase (ESBL)-carrying and colistin-resistant Enterobacteriaceae. AMRB were regularly detected in pig and poultry slaughterhouse wastewaters monitored here. All 25 ESBL-producing bacterial strains (19 E. coli and six K. pneumoniae) isolated from poultry slaughterhouses were multidrug-resistant. In pig slaughterhouses 64% (12 of 21 E. coli [57%] and all four detected K. pneumoniae [100%]) were multidrug-resistant. Regarding colistin, resistant Enterobacteriaceae were detected in 54% of poultry and 21% of pig water samples. Carbapenem resistance was not detected. Resistant bacteria were found directly during discharge of wastewaters from abattoirs into water bodies highlighting the role of slaughterhouses for environmental surface water contamination.
Kinematic Prediction and Experimental Demonstration of Conditioning Process for Controlling the Profile Shape of a Chemical Mechanical Polishing Pad
Authors: Cho Hanchul , Lee Taekyung , Kim Doyeon , Kim Hyoungjae
Published: May 2021

The uniformity of the wafer in a chemical mechanical polishing (CMP) process is vital to the ultra-fine and high integration of semiconductor structures. In particular, the uniformity of the polishing pad corresponding to the tool directly affects the polishing uniformity and wafer shape. In this study, the profile shape of a CMP pad was predicted through a kinematic simulation based on the trajectory density of the diamond abrasives of the diamond conditioner disc. The kinematic prediction was found to be in good agreement with the experimentally measured pad profile shape. Based on this, the shape error of the pad could be maintained within 10 μm even after performing the pad conditioning process for more than 2 h, through the overhang of the conditioner.
Effects of Tea Polyphenol Treatments on the Quality and Microbiota of Crisp Grass Carp Fillets during Storage at 4 °C
Authors: Pan Ziqiang , Li Lin , Shen Zhihua , Chen Yan , Li Mei
Published: May 2021

We investigated the effects of tea polyphenol treatments on the quality and microbiota of crisp grass carp fillets during cold storage at 4 °C. Changes in the total viable count (TVC) and total volatile base nitrogen (TVB-N) of fillets were measured. The microbiota of the crisp grass carp was analyzed using high-throughput sequencing technique. The results indicated that tea polyphenol treatments inhibited bacterial growth and reduced TVB-N values of fish fillets, and subsequently extended their shelf life by 6 days. The high-throughput sequencing result showed that Pseudomonas was the most abundant bacteria in tea polyphenol-treated fish fillets at the end of shelf life, while Pseudomonas and Aeromonas were the most abundant bacteria in control samples. These findings suggested that tea polyphenol treatments could be used in future to extend the shelf life of crisp grass carp fillets and alter the bacterial communities responsible for spoiling fish.
Mitigation of Short-Term Wind Power Ramps through Forecast-Based Curtailment
Authors: Probst Oliver , Minchala Luis I.
Published: May 2021

As the penetration of renewable energy generation in electric grids becomes more substantial, its contribution to the variability of the net load becomes more noticeable. Particularly in small or weak grids, the rate at which the output power of a wind farm decreases may become a concern to grid operators. In the present work, a novel approach, called forecast-based curtailment (FBC), is shown to be able to self-mitigate downward ramps on short time scales at a very small energy penalty, compared to conventional mitigation schemes, such as flat curtailment or up-ramp limitations. FBC allows to achieve compliance with ramp limits imposed by system operators at a very small energy cost and modest additional upfront investments.
Myxofibrosarcoma Mimicking Inflammatory Lesion of Temporomandibular Joint—Case Presentation
Authors: Zagacki Dawid , Sztychny Krzysztof , Tyndorf Marta , Bibik Robert , Sygut Dominik , Kozakiewicz Marcin
Published: May 2021

Treating oncologic patients remains a challenge for surgeons aiming to provide patients with safe margins of resection while maintaining the highest possible quality of life. The latter, in the case of malignancies, requires using sophisticated methods of reconstruction. Thus, we present a case of a 75-year-old patient treated in our department with a rare neoplasm in the region of the temporomandibular joint—a myxofibrosarcoma that was mimicking an inflammatory lesion. The patient underwent two surgeries—firstly alloplasty of the TMJ due to the suspicion of an inflammatory lesion, lately extended to the resection of glenoid fossa and subtemporal fossa contents when the mandible was reconstructed using UHMW-PE (ultra-high molecular weight polyethylene). The patient was also referred for adjuvant radiotherapy and has remained disease-free for over 96 months with very good aesthetics and function of the mandible. The presented case highlights not only the need for increased oncologic awareness but also the possible use of UHMW-PE as a reconstruction material in the broad resection of the maxillofacial region.
Complex Network Modelling of Origin–Destination Commuting Flows for the COVID-19 Epidemic Spread Analysis in Italian Lombardy Region
Authors: Lombardi Angela , Amoroso Nicola , Monaco Alfonso , Tangaro Sabina , Bellotti Roberto
Published: May 2021

Currently the whole world is affected by the COVID-19 disease. Italy was the first country to be seriously affected in Europe, where the first COVID-19 outbreak was localized in the Lombardy region. The further spreading of the cases led to the lockdown of the most affected regions in northern Italy and then the entire country. In this work we investigated an epidemic spread scenario in the Lombardy region by using the origin–destination matrix with information about the commuting flows among 1450 urban areas within the region. We performed a large-scale simulation-based modeling of the epidemic spread over the networks related to three main motivations, i.e., work, study and occasional transfers to quantify the potential contribution of each category of travellers to the spread of the epidemic process. Our findings outline that the three networks are characterised by different weight dynamic growth rates and that the network “work” has a critical role in the diffusion phenomenon showing the greatest contribution to the epidemic spread.
Modeling of Aerodynamic Separation of Preliminarily Stratified Grain Mixture in Vertical Pneumatic Separation Duct
Authors: Kharchenko Serhii , Borshch Yurii , Kovalyshyn Stepan , Piven Mykhailo , Abduev Magomed , Miernik Anna , Popardowski Ernest , Kiełbasa Paweł
Published: May 2021

The productivity of most grain cleaning machines seems to be directly related to the efficiency of vertical pneumatic separation ducts. Nevertheless, improvement is accompanied by an increase in the load of the vertical duct, the design of which is limited by the width of the grain cleaning machines. This requires an increase in the thickness of the layer of grain mixture that enters the working area of the duct, which significantly worsens the conditions of separation of its components under the action of airflow. Particles of light impurities are unable to separate due to their retention by the grain medium. This reduces the quality of cleaning and requires appropriate scientific and technical solutions. The application of preliminary stratification of the granular mixture while increasing the concentration of light impurities in the top layer of the mixture seems to be a prospective method. The positive effect of the previous stratification on the intensity of redistribution of light impurity particles in the working zone has been theoretically considered and experimentally confirmed. Mathematical models were obtained to determine the trajectory of discussed particles, taking into account the previous stratification of mixtures, and the corresponding dependences were established. The influence of the initial coordination of the introduction of the particles of lightweight impurities, their sizes and densities, and technological and structural parameters of operation of the pneumatic separation duct were taken into account in the studies. The intensification of the process of cleaning grain from lightweight impurities in the working zones of pneumatic separation ducts of grain cleaning machines is theoretically substantiated.
Evaluation of Construction Site Noise to Allow the Optimisation of Construction Processes and Construction Machinery Selection
Authors: Kantová Radka
Published: May 2021

During the pre-construction phase of an investment project, a construction plan is usually prepared by the building contractor. It contains financial plans, schedules, technical regulations with working procedures and proposals for the deployment of construction machinery and work teams. These items are then optimised in order to save money, shorten construction time and increase the quality of the final work while ensuring environmental and safety standards are maintained at the site. The ecological aspects of building work include compliance with the hygiene limits for building site noise, which is caused in particular by the simultaneous operation of machines. In the Czech Republic, limit values are set by Government Regulation 272/2011 Coll., as amended, on the protection of health against the adverse effects of noise and vibrations. The evaluation of construction site noise is not usually performed by contractors themselves and noise is usually measured only at the construction site, where work is limited or additional measures are taken in the case of values that exceed the stipulated limits. As the values in the construction plan should be as close as possible to reality during the implementation of the plan, it would seem appropriate to include such noise assessments within it. The purpose of this paper is to present an example of such an evaluation from the formal and material standpoint. The assessment has been conducted in the form of a specific noise study, expanded by a commentary on the treatment of the hypotheses. It shows how to obtain input data for evaluation, divide sub-calculations and use the supporting SW of the Hluk plus product for the purposes of construction information modelling, as well as how to correctly set noise sources and the positions of measured points for evaluation by the model within this programme. Subsequently, options for the evaluation of appropriate measures are listed, including their inclusion in reassessments.
Specific Absolute Velocity Thresholds during Male Basketball Games Using Local Positional System; Differences between Age Categories
Authors: Sosa Carlos , Lorenzo Alberto , Trapero Juan , Ribas Carlos , Alonso Enrique , Jimenez Sergio L.
Published: May 2021

The aim of this study was (I) to establish absolute specific velocity thresholds during basketball games using local positional system (LPS) and (II) to compare the speed profiles between various levels of competitions. The variables recorded were total distance (TD); meters per minute (m·min); real time (min); maximum speed (Km h−1), distance (m), percentage distance, and percentage duration invested in four speed zones (standing–walking; jogging; running; and high-speed running). Mean and standard deviation (±SD) were calculated, and a separate one-way analysis of variance was undertaken to identify differences between competitions. TD (3188.84 ± 808.37 m) is covered by standing–walking (43.51%), jogging (36.58%), running (14.68%), and sprinting (5.23%) activities. Overall, 75.22% of the time is invested standing–walking, jogging (18.43%), running (4.77%), and sprinting (1.89%). M·min (large effect size), % duration zone 2 (moderate effect size); distance zone 4 (large effect size), and % distance zone 4 (very large effect size) are significantly higher during junior than senior. However, % distance zone 1 (large effect size) and % duration zone 1 (large effect size) were largely higher during senior competition. The findings of this study reveal that most of the distance and play time is spent during walking and standing activities. In addition, the proportion of time spent at elevated intensities is higher during junior than in senior competition.
A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes Wound Healing in a Primary Gingival Fibroblast Model
Authors: Colangelo Maria Teresa , Belletti Silvana , Govoni Paolo , Guizzardi Stefano , Galli Carlo
Published: May 2021

Polynucleotides (PN) have long been known as an effective supportive therapy for wound healing. The present study investigated whether a hydrogel formulation containing PN and hyaluronic acid (PN + HA) could promote wound healing in an in vitro model of gingival fibroblasts. PN promoted cell growth and viability as assessed by different assays, and PN + HA, though not significantly further increasing cell growth as compared to PN, supported the formation of dense multilayered cell nodules. PN promoted fibroblasts’ clonogenic efficiency and PN + HA further enhanced the formation of more numerous dense colonies. PN + HA appeared to significantly increase the expression of collagen 1a1 and 3a1, while not affecting proteoglycans deposition. Interestingly, when tested in a scratch assay, PN + HA achieved gap closure after 48 h, while cells in the comparison groups had not completely bridged the scratch even after 96 h. Taken together, these results demonstrate that PN + HA is a promising candidate for a supportive therapy to promote soft tissue healing in the oral cavity.
Sperm Selection by Magnetic-Activated Cell Sorting before Microinjection of Autologous Oocytes Increases Cumulative Live Birth Rates with Limited Clinical Impact: A Retrospective Study in Unselected Males
Authors: Gil Juliá María , Hervás Irene , Navarro-Gómez Lechón Ana , Quintana Fernando , Amorós David , Pacheco Alberto , González-Ravina Cristina , Rivera-Egea Rocío , Garrido Nicolás
Published: May 2021

The application of MACS non-apoptotic sperm selection in infertility clinics is controversial since the published literature does not agree on its effect on reproductive outcomes. Therefore, it is not part of the routine clinical practice. Classical measures of reproductive success (pregnancy or live birth rates per ovarian stimulation) introduce a bias in the evaluation of a technique’s effect, since only the best embryo is transferred. This retrospective, multicenter, observational study evaluated the impact of MACS on reproductive outcomes, measuring results in classical parameters and cumulative live birth rates (CLBR). Data from ICSI cycles using autologous oocyte in Spanish IVIRMA fertility clinics from January 2008 to February 2020 were divided into two groups according to their semen processing: standard practice (reference: 46,807 patients) versus an added MACS sperm selection (1779 patients). Only when measured as CLBR per embryo transferred and per MII oocyte used was the difference between groups statistically significant. There were no significant differences between MACS and reference groups on pregnancy and live birth rates. In conclusion, results suggest that non-apoptotic sperm selection by MACS on unselected males prior to ICSI with autologous oocytes has limited clinical impact, showing a subtle increase in CLBR per embryo transferred.
Cytotoxic Compounds of Two Demosponges (Aplysina aerophoba and Spongia sp.) from the Aegean Sea
Authors: Orfanoudaki Maria , Hartmann Anja , Alilou Mostafa , Mehic Naida , Kwiatkowski Marcel , Jöhrer Karin , Nguyen Ngoc Hieu , Hensel Andreas , Greil Richard , Ganzera Markus
Published: May 2021

The class of demosponges is the biggest and most diverse of all described sponge species and it is reported to produce a plethora of chemically different metabolites with interesting biological activities. The focus of the present study was to investigate the chemical composition of two Mediterranean demosponges, targeting their brominated compounds and prenylated hydroquinones, compounds with interesting cytotoxic and anti-microbial properties. In order to gain a deeper insight into the chemical diversity of their metabolites and their activities, 20 pure secondary metabolites including new natural products were isolated from two different species (Aplysina aerophoba and Spongia sp.) using various chromatographic techniques. Their structures were confirmed by NMR and HRMS, revealing molecules with various chemical scaffolds, mainly prenylated hydroquinones from Spongia sp. and halogenated compounds from Aplysina aerophoba, including 5 novel natural products. The isolated compounds were investigated for their cytotoxic properties using 9 different cell lines, and especially one compound, 2,6-dibromo-4-hydroxy-4-methoxycarbonylmethylcyclohexa-2,5-dien-1-one showed good activities in all tested models.
Investigating the Relationship between Autistic Traits, Ruminative Thinking, and Suicidality in a Clinical Sample of Subjects with Bipolar Disorder and Borderline Personality Disorder
Authors: Dell’Osso Liliana , Cremone Ivan Mirko , Amatori Giulia , Cappelli Andrea , Cuomo Alessandro , Barlati Stefano , Massimetti Gabriele , Vita Antonio , Fagiolini Andrea , Carmassi Claudia , Carpita Barbara
Published: May 2021

(1) Background: Previous literature reported in both subjects with Borderline personality disorder (BPD) and Bipolar disorder (BD) higher levels of autistic traits, linked to a greater suicidality risk. The aim of this study was to evaluate and compare the presence of autistic traits in a sample of individuals with BD or BPD, with a specific focus on suicidality. (2) Methods: We recruited two clinical samples of subjects (BPD and BD) and a control group without a diagnosis according to DSM-5 (CTL). Subjects were assessed with the AdAS Spectrum, the RRS and, for evaluating suicidality, the MOODS-SR. (3) Results: The CTL group showed significantly lower scores of both BD and BPD on AdAS Spectrum, RRS, and suicidality scores. BPD subjects showed significantly lower scores than BD ones in most of AdAS Spectrum domain scores. Correlation and regression analyses highlighted specific patterns of association among AdAS Spectrum domains, RRS, and suicidality in each clinical group. (4) Conclusions: Both BPD and BD individuals show greater levels of autistic traits, which seem to be distributed in a continuum featuring the highest levels among BD subjects. In both disorders, higher autistic traits were linked to suicidal tendencies, although with different patterns of association between BD and BPD subjects.
Biomarkers of Oncogenesis, Adipose Tissue Dysfunction and Systemic Inflammation for the Detection of Hepatocellular Carcinoma in Patients with Nonalcoholic Fatty Liver Disease
Authors: Caviglia Gian Paolo , Armandi Angelo , Rosso Chiara , Gaia Silvia , Aneli Serena , Rolle Emanuela , Abate Maria Lorena , Olivero Antonella , Nicolosi Aurora , Guariglia Marta , Ribaldone Davide Giuseppe , Carucci Patrizia , Saracco Giorgio Maria , Bugianesi Elisabetta
Published: May 2021

Current surveillance strategy for patients with nonalcoholic fatty liver disease (NAFLD) at risk of hepatocellular carcinoma (HCC) development is unsatisfactory. We aimed to investigate the diagnostic accuracy of alpha-fetoprotein (AFP), protein induced by vitamin K absence or antagonist-II (PIVKA-II), glypican-3 (GPC-3), adiponectin, leptin and interleukin-6 (IL-6), alone or in combination, for the discrimination between NAFLD patients with or without HCC. The biomarkers were investigated in a cohort of 191 NAFLD patients (median age 62, 54–68 years; 121 males and 70 females) with advanced fibrosis/cirrhosis, 72 of whom had a diagnosis of HCC. PIVKA-II showed the best performance for the detection of HCC with an area under the curve (AUC) of 0.853, followed by adiponectin (AUC = 0.770), AFP (AUC = 0.763), GPC-3 (AUC = 0.759) and by IL-6 (AUC = 0.731), while the leptin values were not different between patients with and without HCC. The accuracy of the biomarkers’ combination was assessed by a stratified cross-validation approach. The combination of age, gender, PIVKA-II, GPC-3 and adiponectin further improved the diagnostic accuracy (AUC = 0.948); the model correctly identified the 87% of the patients. In conclusion, we developed a model with excellent accuracy for the detection of HCC that may be useful to improve the surveillance of NAFLD patients at risk of tumor development.
Empirical Evaluation of the Use of Computational HLA Binding as an Early Filter to the Mass Spectrometry-Based Epitope Discovery Workflow
Authors: Bouzid Rachid , de Beijer Monique T. A. , Luijten Robbie J. , Bezstarosti Karel , Kessler Amy L. , Bruno Marco J. , Peppelenbosch Maikel P. , Demmers Jeroen A. A. , Buschow Sonja I.
Published: May 2021

Immunopeptidomics is used to identify novel epitopes for (therapeutic) vaccination strategies in cancer and infectious disease. Various false discovery rates (FDRs) are applied in the field when converting liquid chromatography-tandem mass spectrometry (LC-MS/MS) spectra to peptides. Subsequently, large efforts have recently been made to rescue peptides of lower confidence. However, it remains unclear what the overall relation is between the FDR threshold and the percentage of obtained HLA-binders. We here directly evaluated the effect of varying FDR thresholds on the resulting immunopeptidomes of HLA-eluates from human cancer cell lines and primary hepatocyte isolates using HLA-binding algorithms. Additional peptides obtained using less stringent FDR-thresholds, although generally derived from poorer spectra, still contained a high amount of HLA-binders and confirmed recently developed tools that tap into this pool of otherwise ignored peptides. Most of these peptides were identified with improved confidence when cell input was increased, supporting the validity and potential of these identifications. Altogether, our data suggest that increasing the FDR threshold for peptide identification in conjunction with data filtering by HLA-binding prediction, is a valid and highly potent method to more efficient exhaustion of immunopeptidome datasets for epitope discovery and reveals the extent of peptides to be rescued by recently developed algorithms.
Unmasking BCL-2 Addiction in Synovial Sarcoma by Overcoming Low NOXA
Authors: Fairchild Carter K. , Floros Konstantinos V. , Jacob Sheeba , Coon Colin M. , Puchalapalli Madhavi , Hu Bin , Harada Hisashi , Dozmorov Mikhail G. , Koblinski Jennifer E. , Smith Steven C. , Domson Gregory , Leverson Joel D. , Souers Andrew J. , Takebe Naoko , Ebi Hiromichi , Faber Anthony C. , Boikos Sosipatros A.
Published: May 2021

Synovial sarcoma (SS) is frequently diagnosed in teenagers and young adults and continues to be treated with polychemotherapy with variable success. The SS18-SSX gene fusion is pathognomonic for the disease, and high expression of the anti-apoptotic BCL-2 pathologically supports the diagnosis. As the oncogenic SS18-SSX fusion gene itself is not druggable, BCL-2 inhibitor-based therapies are an appealing therapeutic opportunity. Venetoclax, an FDA-approved BCL-2 inhibitor that is revolutionizing care in some BCL-2-expressing hematological cancers, affords an intriguing therapeutic possibility to treat SS. In addition, there are now dozens of venetoclax-based combination therapies in clinical trials in hematological cancers, attributing to the limited toxicity of venetoclax. However, preclinical studies of venetoclax in SS have demonstrated an unexpected ineffectiveness. In this study, we analyzed the response of SS to venetoclax and the underlying BCL-2 family biology in an effort to understand venetoclax treatment failure and find a therapeutic strategy to sensitize SS to venetoclax. We found remarkably depressed levels of the endogenous MCL-1 inhibitor, NOXA, in SS compared to other sarcomas. Expressing NOXA led to sensitization to venetoclax, as did the addition of the MCL-1 BH3 mimetic, S63845. Importantly, the venetoclax/S63845 combination induced tumor regressions in SS patient-derived xenograft (PDX) models. As a very close analog of S63845 (S64315) is now in clinical trials with venetoclax in AML (NCT03672695), the combination of MCL-1 BH3 mimetics and venetoclax should be considered for SS patients as a new therapy.
Circulating Cell-Free DNA as a Prognostic Biomarker in Resectable Ampullary Cancer
Authors: Shyr Bor-Uei , Shyr Bor-Shiuan , Chen Shih-Chin , Chang Shih-Ching , Shyr Yi-Ming , Wang Shin-E
Published: May 2021

Circulating cell-free DNA (cfDNA) in ampullary cancer patients was measured to clarify the correlation between cfDNA and clinicopathological factors and the impact of cfDNA on survival outcomes. Patients with ampullary cancer undergoing pancreaticoduodenectomy were included. Correlations between cfDNA and clinicopathological and prognostic factors were determined. The cfDNA levels in patients ranged from 1282 to 21,674 copies/mL, with a median of 6687 copies/mL. The cfDNA level was significantly higher in patients with lymph node involvement, lymphovascular invasion, abnormal serum carcinoembryonic antigen (CEA) level, and stage II and III cancer. Poor prognostic factors for ampullary cancer included high cfDNA > 6687 copies/mL, lymph node involvement, abnormal serum CEA > 5 ng/mL, and advanced stage II and III cancer. The 1- and 5-year survival rates were 92.0% and 66.5%, respectively, for patients with low cfDNA < 6687 copies/mL and 84.0% and 49.9%, respectively, for patients with high cfDNA > 6687 copies/mL (p < 0.001). After multivariate analysis, only the cfDNA level and stage were independent prognostic factors of ampullary cancer. Thus, the cfDNA level could act as a surrogate marker of both disease extent and biological aggressiveness of ampullary cancer. Moreover, cfDNA plays a significant role in the prognosis of resectable ampullary cancer.
Validation of the Combined Biomarker for Prediction of Response to Checkpoint Inhibitor in Patients with Advanced Cancer
Authors: Kim Jin-Chul , Heo You-Jeong , Kang So-Young , Lee Jeeyun , Kim Kyoung-Mee
Published: May 2021

Although immune checkpoint inhibitors can induce durable responses in patients with multiple types of advanced cancer, only a limited number of patients have a known reliable biomarker. This study aimed to validate the IMmunotherapy Against GastrIc Cancer (IMAGiC) model, which was developed based on a previous study of four-gene and PD-L1 level, to predict immunotherapy response. We developed a clinical assay for formalin-fixed paraffin-embedded samples using quantitative real-time polymerase chain reaction to measure the expression level of the previously published four-gene set. The predictive performance was validated in a cohort of 89 patients with several advanced tumor types. The IMAGiC score was derived from tumor samples of 89 patients consisting of eight cancer types, and 73 out of 89 patients available for clinical response were analyzed with clinicopathological factors. The IMAGiC group (responder vs. non-responder) was determined with a specific value of the IMAGiC score as a cutoff, which was set by log-rank statistics for progression-free survival (PFS) divided the patients into 56 (76.7%) non-responders and 17 (23.3%) responders. Clinical responders (complete response/partial response) were higher in the IMAGiC responder group than in the non-responder group (70.6 vs. 21.4%). The median PFS of the IMAGiC responder group and non-responder was 20.8 months (95% CI 9.1-not reached) and 6.7 months (95% CI 4.9–11.1, p = 0.007), respectively. Among the 17 IMAGiC responders, 11 patients had tumor mutation burden-low and microsatellite-stable tumors. This study validated a predictive model based on a four-gene expression signature. Along with conventional biomarkers, our model could be useful for predicting response to immunotherapy in patients with advanced cancer.
Brain Tumor Resection in Elderly Patients: Potential Factors of Postoperative Worsening in a Predictive Outcome Model
Authors: Ferroli Paolo , Vetrano Ignazio Gaspare , Schiavolin Silvia , Acerbi Francesco , Zattra Costanza Maria , Schiariti Marco , Leonardi Matilde , Broggi Morgan
Published: May 2021

The decision of whether to operate on elderly patients with brain tumors is complex, and influenced by pathology-related and patient-specific factors. This retrospective cohort study, based on a prospectively collected surgical database, aims at identifying possible factors predicting clinical worsening after elective neuro-oncological surgery in elderly patients. Therefore, all patients ≥65 years old who underwent BT resection at a tertiary referral center between 01/2018 and 12/2019 were included. Age, smoking, previous radiotherapy, hypertension, preoperative functional status, complications occurrence, surgical complexity and the presence of comorbidities were prospectively collected and analyzed at discharge and the 3-month follow-up. The series included 143 patients (mean 71 years, range 65–86). Sixty-five patients (46%) had at least one neurosurgical complication, whereas 48/65 (74%) complications did not require invasive treatment. Forty-two patients (29.4%) worsened at discharge; these patients had a greater number of complications compared to patients with unchanged/improved performance status. A persistent worsening at three months of follow-up was noted in 20.3% of patients; again, this subgroup presented more complications than patients who remained equal or improved. Therefore, postoperative complications and surgical complexity seem to influence significantly the early outcome in elderly patients undergoing brain tumor surgery. In contrast, postoperative complications alone are the only factor with an impact on the 3-month follow-up.
Utility and Mechanism of SHetA2 and Paclitaxel for Treatment of Endometrial Cancer
Authors: Chandra Vishal , Rai Rajani , Benbrook Doris Mangiaracina
Published: May 2021

Endometrial cancer patients with advanced disease or high recurrence risk are treated with chemotherapy. Our objective was to evaluate the utility and mechanism of a novel drug, SHetA2, alone and in combination with paclitaxel, in endometrial cancer. SHetA2 targets the HSPA chaperone proteins, Grp78, hsc70, and mortalin, which have high mutation rates in endometrial cancer. SHetA2 effects on cancerous phenotypes, mitochondria, metabolism, protein expression, mortalin/client protein complexes, and cell death were evaluated in AN3CA, Hec13b, and Ishikawa endometrial cancer cell lines, and on growth of Ishikawa xenografts. In all three cell lines, SHetA2 inhibited anchorage-independent growth, migration, invasion, and ATP production, and induced G1 cell cycle arrest, mitochondrial damage, and caspase- and apoptosis inducing factor (AIF)-mediated apoptosis. These effects were associated with altered levels of proteins involved in cell cycle regulation, mitochondrial function, protein synthesis, endoplasmic reticulum stress, and metabolism; disruption of mortalin complexes with mitochondrial and metabolism proteins; and inhibition of oxidative phosphorylation and glycolysis. SHetA2 and paclitaxel exhibited synergistic combination indices in all cell lines and exerted greater xenograft tumor growth inhibition than either drug alone. SHetA2 is active against endometrial cancer cell lines in culture and in vivo and acts synergistically with paclitaxel.
Assessment of PI3K/mTOR/AKT Pathway Elements to Serve as Biomarkers and Therapeutic Targets in Penile Cancer
Authors: Thomas Anita , Reetz Sascha , Stenzel Philipp , Tagscherer Katrin , Roth Wilfried , Schindeldecker Mario , Michaelis Martin , Rothweiler Florian , Cinatl Jindrich , Cinatl Jaroslav , Dotzauer Robert , Vakhrusheva Olesya , Albersen Maarten , Macher-Goeppinger Stephan , Haferkamp Axel , Juengel Eva , Neisius Andreas , Tsaur Igor
Published: May 2021

The PI3K/mTOR/AKT pathway might represent an intriguing option for treatment of penile cancer (PeCa). We aimed to assess whether members of this pathway might serve as biomarkers and targets for systemic therapy. Tissue of primary cancer from treatment-naïve PeCa patients was used for tissue microarray analysis. Immunohistochemical staining was performed with antibodies against AKT, pAKT, mTOR, pmTOR, pS6, pPRAS, p4EBP1, S6K1 and pp70S6K. Protein expression was correlated with clinicopathological characteristics as well as overall survival (OS), disease-specific survival (DSS), recurrence-free survival (RFS) and metastasis-free survival (MFS). AKT inhibition was tested in two primarily established, treatment-naïve PeCa cell lines by treatment with capivasertib and analysis of cell viability and chemotaxis. A total of 76 patients surgically treated for invasive PeCa were included. Higher expression of AKT was significantly more prevalent in high-grade tumors and predictive of DSS and OS in the Kaplan–Meier analysis, and an independent predictor of worse OS and DSS in the multivariate regression analysis. Treatment with pan-AKT inhibitor capivasertib in PeCa cell lines induced a significant downregulation of both total AKT and pAKT as well as decreased cell viability and chemotaxis. Selected protein candidates of the mTOR/AKT signaling pathway demonstrate association with histological and survival parameters of PeCa patients, whereas AKT appears to be the most promising one.
LIMK2-NKX3.1 Engagement Promotes Castration-Resistant Prostate Cancer
Authors: Sooreshjani Moloud A. , Nikhil Kumar , Kamra Mohini , Nguyen Dung N. , Kumar Dinesh , Shah Kavita
Published: May 2021

NKX3.1’s downregulation is strongly associated with prostate cancer (PCa) initiation, progression, and CRPC development. Nevertheless, a clear disagreement exists between NKX3.1 protein and mRNA levels in PCa tissues, indicating that its regulation at a post-translational level plays a vital role. This study identified a strong negative relationship between NKX3.1 and LIMK2, which is critical in CRPC pathogenesis. We identified that NKX3.1 degradation by direct phosphorylation by LIMK2 is crucial for promoting oncogenicity in CRPC cells and in vivo. LIMK2 also downregulates NKX3.1 mRNA levels. In return, NKX3.1 promotes LIMK2’s ubiquitylation. Thus, the negative crosstalk between LIMK2-NKX3.1 regulates AR, ARv7, and AKT signaling, promoting aggressive phenotypes. We also provide a new link between NKX3.1 and PTEN, both of which are downregulated by LIMK2. PTEN loss is strongly linked with NKX3.1 downregulation. As NKX3.1 is a prostate-specific tumor suppressor, preserving its levels by LIMK2 inhibition provides a tremendous opportunity for developing targeted therapy in CRPC. Further, as NKX3.1 downregulates AR transcription and inhibits AKT signaling, restoring its levels by inhibiting LIMK2 is expected to be especially beneficial by co-targeting two driver pathways in tandem, a highly desirable requisite for developing effective PCa therapeutics.
The Role of the Microbiome in Liver Cancer
Authors: Moreno-Gonzalez Mar , Beraza Naiara
Published: May 2021

Hepatocellular carcinoma (HCC) is the most common malignancy occuring in the context of chronic liver disease and is one of the main causes of cancer-derived death worldwide. The lack of effective treatments, together with the poor prognosis, underlines the urge to develop novel and multidisciplinary therapeutics. An increasing body of evidence shows that HCC associates with changes in intestinal microbiota abundance and composition as well as with impaired barrier function, leading to the release of bacteria and their metabolites to the liver. These factors trigger a cascade of inflammatory responses contributing to liver cirrhosis and constituting an ideal environment for the progression of HCC. Interestingly, the use of bacteriotherapy in human and preclinical studies of chronic liver disease and HCC has been shown to successfully modify the microbiota composition, reducing overall inflammation and fibrosis. In this review, we explore the existing knowledge on the characterisation of the intestinal microbial composition in humans and experimental murine chronic liver disease and HCC, as well as the use of antibiotics and bacteriotherapy as therapeutic options.