bluuebunny/crossref_metadata_2025 huggingface dataset:
Implications of Permanent Teeth Dimensions and Arch Lengths on Dental Crowding during the Mixed Dentition Period
Authors: Daoud Raisa , Bencze Maria-Angelica , Albu Cristina-Crenguța , Teodorescu Elina , Dragomirescu Anca-Oana , Vasilache Adriana , Suciu Ioana , Ionescu Ecaterina
Published: August 2021

Dento-alveolar disharmony with crowding is a common reason for orthodontic treatment with not fully understood or unequivocally demonstrated causes. This study investigated the correlations between teeth dimensions, arch lengths, and crowding during the mixed dentition period. A cross-sectional study on 100 dental casts of patients with class I malocclusions was performed. Dental arches were classified as non-crowded, moderately crowded, severely crowded, and spaced. The mesio-distal widths, bucco-lingual sizes, and crown proportions of permanent teeth were assessed. The results indicated that arch length measurements showed smaller values in crowded arches. The mesio-distal dimensions of upper central incisors and lower lateral incisors were larger in patients with crowding. The bucco-lingual dimensions of upper incisors were decreased, the bucco-lingual dimensions of lower central incisors and permanent first molars were increased in crowded arches. Upper incisors and lower lateral incisors presented larger crown proportions in crowding cases. Low negative correlations were found between mesio-distal diameters of maxillary central incisors, lower lateral incisors, lower permanent first molars, and the values of arch space discrepancies. In conclusion, crowding in the mixed dentition could be associated with reduced arch lengths, increase in mesio-distal sizes of incisors and lower permanent first molars, and variations of bucco-lingual dimensions and crown proportions of incisors and lower permanent first molars.
Aromatic Profile of Hydroponically and Conventionally Grown Tomatoes
Authors: Korčok Melina , Vietorisová Nikola , Martišová Patrícia , Štefániková Jana , Mravcová Anna , Vietoris Vladimír
Published: August 2021

Hydroponics is a more environmentally friendly and economical way of growing crops that allows crops to be grown all year round, regardless of soil and climate conditions. Hydroponic cultivation of various fruits, vegetables, flowers, etc., is well known and used today. Tomatoes also play an important role in hydroponic cultivation. Tomatoes grown in this way should have a more pronounced aroma and flavor, and the fact that they are harvested at the ripe stage should also be reflected in the fresh red color and appropriate organoleptic characteristics of the fruit. This study was concerned with the sensory analysis of hydroponically grown tomatoes (Solanum lycopersicum) compared with conventionally grown tomatoes of the same species. Samples were evaluated by instrumental sensory analysis using an electronic nose as well as a sensory panel. In this study, the difference between hydroponically and conventionally grown tomatoes was demonstrated. These differences were also captured by the consumers (sensory panel). When analyzing the odor profile of the samples, we found that hydroponic tomatoes are characterized by the presence of chemical organic compounds, namely: 2-methylpropanol, 2,3-pentanedione, and (Z)-3-hexen-1-ol or 1-hexanol, which cause the fruity aroma of the fruit. These substances are very likely to characterize the differences between the tested samples. The electronic nose has shown to be a potentially suitable tool for detecting differences and identifying typical product markers, which may suggest its further use in food authentication detection. It is also interesting to find almost no correlation between the two methods studied.
Toxic Effects of Heavy Metals and Organic Polycyclic Aromatic Hydrocarbons in Sediment Porewater on the Amphipod Hyalella azteca and Zebrafish Brachydanio rerio Embryos from Different Rivers in Taiwan
Authors: Hu Shao-Yang , Hsieh Chi-Ying , Dahms Hans-Uwe , Tseng Yu-Hsien , Chen Jesse , Wu Meng-Chun , Kim Jin-Hyoung , Liu Cheng-Han
Published: August 2021

The amphipod (Hyalella azteca) and zebrafish (Brachydanio rerio) embryos were used for toxicological sediment porewater testing. Porewaters from 35 sampling stations of eight streams in southern Taiwan were screened for toxic effects and their relationship with 6 metal and 16 polycyclic aromatic hydrocarbons (PAHs). Concentration analysis results showed that the following PAHs, naphththalene, benzo(b)fluoranthene, dibenz(a,h)anthracene, acenaphthalene, and the heavy metal cadmium were not detected in 35 sampling stations. The highest detection rate of 94.1% was caused by the PAHs fluoranthene and pyrene. The highest detection rate of the metal zinc was 88.6% of 35 analyzed samples. The majority of samples (88%) were classified as level tier 1 according to USEPA national sediment inventory. This indicates the probability of adverse effects on aquatic life or human health. The results of a zebrafish embryo test showed that heart rate and survival were significantly reduced with all porewater samples. Therefore, fish exposed to contaminated river conditions may be affected in their cardiovascular functions. Looking at correlations between toxic effects of metals and PAHs, we found that phenanthrene, anthracene, pyrene, benzo(a)anthracene, chrysene, benzo(b)fluoranthene, and benzo(a)pyrene were low, while fluorene was highly correlated with toxic effects of metals.
Effects of Pitch Size and Goalkeeper Participation on Physical Load Measures during Small-Sided Games in Sub-Elite Professional Soccer Players
Authors: Santos Fernando J. , Verardi Carlos E. L. , de Moraes Mayra Grava , Filho Dalton M. Pessôa , Macedo Anderson G. , Figueiredo Teresa P. , Ferreira Cátia C. , Borba Ricardo P. , Espada Mário C.
Published: August 2021

This study sought to examine the variations of internal and external load measures in soccer players when performing different types of 4 vs. 4 small-sided games (SSGs). Ten U-23 professional and sub-elite soccer players, eight outfield players (age: 20.1 ± 1.5 years old; height: 1.83 ± 0.04 m; total body mass: 76.1 ± 3.6 kg) and two goalkeepers (GK) (age: 20.5 ± 2.1 years old; height: 1.91 ± 0.04 m; total body mass: 77.0 ± 1.4 kg) participated in the study. Five SSGs were performed with 3 min duration and 3 min rest interval. SSG1, SSG2 and SSG4 aimed for ball possession play, while in SSG3 and SSG5, two GK were added, defending regular-size goals. The pitch size changed from SSG1 (16 × 24 m) to SSG2 and SSG3 (both 20 × 30 m) and SSG4 and SSG5 (both 24 × 36 m). All players were monitored using WIMU Pro device during the tasks. Anova One-Way and the post hoc Tukey honestly significant difference were used for analysis of statistically significant differences (p < 0.05) and effect sizes were determined by calculating partial eta-squared. The framework was associated with relevant but steady state internal load fatigue and induce a considerable external load stimulus. GK participation in SSGs should be carefully considered as it seems to have a greater impact on physical load measures when compared to the pitch size manipulation.
Condition Monitoring Method for the Detection of Fault Graduality in Outer Race Bearing Based on Vibration-Current Fusion, Statistical Features and Neural Network
Authors: Saucedo-Dorantes Juan-Jose , Zamudio-Ramirez Israel , Cureno-Osornio Jonathan , Osornio-Rios Roque Alfredo , Antonino-Daviu Jose Alfonso
Published: August 2021

Bearings are the elements that allow the rotatory movement in induction motors, and the fault occurrence in these elements is due to excessive working conditions. In induction motors, electrical erosion remains the most common phenomenon that damages bearings, leading to incipient faults that gradually increase to irreparable damages. Thus, condition monitoring strategies capable of assessing bearing fault severities are mandatory to overcome this critical issue. The contribution of this work lies in the proposal of a condition monitoring strategy that is focused on the analysis and identification of different fault severities of the outer race bearing fault in an induction motor. The proposed approach is supported by fusion information of different physical magnitudes and the use of Machine Learning and Artificial Intelligence. An important aspect of this proposal is the calculation of a hybrid-set of statistical features that are obtained to characterize vibration and stator current signals by its processing through domain analysis, i.e., time-domain and frequency-domain; also, the fusion of information of both signals by means of the Linear Discriminant Analysis is important due to the most discriminative and meaningful information is retained resulting in a high-performance condition characterization. Besides, a Neural Network-based classifier allows validating the effectiveness of fusion information from different physical magnitudes to face the diagnosis of multiple fault severities that appear in the bearing outer race. The method is validated under an experimental data set that includes information related to a healthy condition and five different severities that appear in the outer race of bearings.
The Lunette with St. Thomas Becket at the Sacro Speco in Subiaco: An Unexpected Presence?
Authors: Cerone Roberta
Published: August 2021

The painting with St. Thomas Becket, St. Stephen and St. Nicholas of Bari that decorates one of the lunettes in the so-called lower church at the Sacro Speco in Subiaco is an enigma from an art-historical point of view, for two reasons. First, on an iconographical level, the lunette interrupts the flow of the story of Benedict’s life unfolding systematically on all the walls of the lower church. Second, from the formal point of view, the fresco clearly presents more archaic features than the surrounding Stories of St. Benedict, dating to the end of the thirteenth/beginning of the fourteenth century, and was therefore probably executed in a phase prior to the cycle of Benedict. In the paper, therefore, I will analyse the motivations that led to the preservation of this painting when the hall was renovated and later redecorated in the late thirteenth century, and discuss the hypotheses surrounding patronage. Both aspects will help to better contextualize the reasons for the presence of the image of St. Thomas Becket in a pre-eminent position in the sanctuary of Benedict in Subiaco, a papal bulwark on the borders of the Kingdom.
A Simple Multi-Parameter Method for Efficient Charging Scheduling of Electric Vehicles
Authors: Konstantinidis George , Kanellos Fotios , Kalaitzakis Kostas
Published: August 2021

In this article, a method for the efficient charging of electric vehicles (EVs) at the parking lot (PL) level, including V2G operation and taking into account lifetime of EV batteries, distribution network and local transformer loading, is proposed. The main targets of the method are to minimize the total charging cost of the PLs hosting the EVs and to satisfy all technical and operation constraints of EVs and PLs. The proposed method exploits particle swarm optimization (PSO) to derive the charging schedule of the EVs. The proposed method is compared with conventional charging strategies, where the EVs are charged with the maximum power of their charging power converter or the average power required to achieve their state-of-charge target, and a conventional charging scheduling method using the aggregated behavior of the plug-in EVs. Real-world data series of electricity price and parking lot activity were used. The results obtained from the study of indicative operation scenarios prove the effectiveness of the proposed method, while no sophisticated computing, measurement and communication systems are required for its application.
Analysis of Marine Diesel Engine Emission Characteristics of Different Power Ranges in China
Authors: Ma Zhongmin , Yang Yuanyuan , Sun Peiting , Xing Hui , Duan Shulin , Qu Hongfei , Zou Yongjiu
Published: August 2021

In order to accurately assess China’s port air pollution caused by the shipping industry, two main methods can be used to calculate the emissions of ships, including the method based on ship fuel consumption and the method based on ship activities. Both methods require accurate diesel engine emission factors, or specific emissions. In this paper, the emission characteristics of NOX, CO, CO2 and THC from 197 domestic marine diesel engines were tested under bench test conditions by a standard emission measurement system. The diesel engines were divided into six Classes, A~F, according to their power distribution, and the fuel-based emission factors and energy-based emission factors of marine main engine and auxiliary engine meeting IMO NOX Tier II standards were given. The results showed that the main engine fuel-based emission factors of NOX, CO, CO2 and THC from Class A to Class F were 33.25~76.58, 2.70~4.33, 3123.92~3166.47 and 1.10~2.64 kg/t-fuel, respectively; and the energy-based emission factors were 6.57~11.75, 0.56~0.81, 530.28~659.71 and 0.18~0.61 g/kW h, respectively. The auxiliary engine fuel-based emission factors of NOX, CO, CO2 and THC from Class A to Class D were 27.17~39.81, 2.66~5.12, 3113.01~3141.34 and 1.16~2.87 kg/t-fuel respectively; and their energy-based emission factors were 6.06~8.33, 0.47~0.77, 656.86~684.91 and 0.21~0.61 g/kW h, respectively. The emission factors for different types of diesel engines were closely related to the diesel engine load, and the relation between them could be expressed by quadratic polynomial or power function. The results of this paper provide valuable data for the estimation of waterway transportation exhaust emissions and comprehensive understanding of the emission characteristics of marine diesel engines.
Research and Optimization of Meteo-Particle Model for Wind Retrieval
Authors: Zhu Jiahui , Wang Haijiang , Li Jing , Xu Zili
Published: August 2021

As the aviation industry has entered a critical period of development, the demand for Automatic Dependent Surveillance Broadcast (ADS-B) technology is becoming increasingly urgent. Real-time detection of aviation wind field information and the early warning of wind field shear by atmospheric sounding system are two important factors related to the safe operation of aviation and airport. According to the advantages of ADS-B and Mode S data, this paper uses the Meteo-Particle (MP) model proposed by Sun et al., in their previous research to retrieve high-altitude wind field. Comparing the precision and accuracy of wind field retrieved results, and the optimization parameters of MP model suitable for meteorological model are further studied. To solve the problem of incomplete wind field coverage obtained by retrieval, an extrapolation algorithm of wind field is proposed. The results show that: (1) a comprehensive evaluation index is introduced, which can more effectively evaluate the comprehensive difference of wind field retrieval results in wind speed and direction. (2) The adaptability results of MP model in different periods and altitudes provide some reference for the research of other scholars. (3) The new parameter setting can improve the accuracy of the retrieved results, and the appropriate extrapolation of wind field fills in the blank part of aviation and meteorology.
Relativistic Study on the Scattering of e± from Atoms and Ions of the Rn Isonuclear Series
Authors: Khandker Mahmudul H. , Haque A. K. Fazlul , Haque M. M. , Billah M. Masum , Watabe Hiroshi , Uddin M. Alfaz
Published: August 2021

Calculations are presented for differential, integrated elastic, momentum transfer, viscosity, inelastic, total cross sections and spin polarization parameters S, T and U for electrons and positrons scattering from atoms and ions of radon isonuclear series in the energy range from 1 eV–1 MeV. In addition, we analyze systematically the details of the critical minima in the elastic differential cross sections along with the positions of the corresponding maximum polarization points in the Sherman function for the aforesaid scattering systems. Coulomb glory is investigated across the ionic series. A short range complex optical potential, comprising static, polarization and exchange (for electron projectile) potentials, is used to describe the scattering from neutral atom. This potential is supplemented by the Coulomb potential for the same purpose for a charged atom. The Dirac partial wave analysis, employing the aforesaid potential, is carried out to calculate the aforesaid scattering observables. A comparison of our results with other theoretical findings shows a reasonable agreement over the studied energy range.
Evaluation of Fe XIV Intensity Ratio for Electron Density Diagnostics by Laboratory Measurements
Authors: Kambara Nagaaki , Kawate Tomoko , Oishi Tetsutarou , Kawamoto Yasuko , Sakaue Hiroyuki A. , Kato Daiji , Nakamura Nobuyuki , Hara Hirohisa , Murakami Izumi
Published: August 2021

The intensity ratio of Fe XIV 264.765A/274.203A is useful to determine the electron density of solar corona, and the relationship between the electron density and the intensity ratio obtained from a model should be evaluated using laboratory plasmas to estimate the electron density more precisely. We constructed a new collisional–radiative model (CR-model) for Fe XIV (an Al-like iron ion) by considering the processes of proton-impact excitation and electron-impact ionization to the excited states of a Mg-like iron ion. The atomic data used in the CR-model were calculated using the HULLAC atomic code. The model was evaluated based on laboratory experiments using a compact electron beam ion trap, called CoBIT, and the Large Helical Device (LHD). The measured Fe XIV 264.785 Å/274.203 Å line intensity ratio with CoBIT was 1.869 ± 0.036, and it agreed well with our CR-model results. Concurrently, the measured ratio using LHD was larger than the results of our CR-model and CHIANTI. The estimated electron densities using our CR-model agreed with those from CHIANTI within a factor of 1.6–2.4 in the range of ne≈1010−11cm−3. Further model development is needed to explain the ratio in a high-electron density region.
A Parametric Type of Cauchy Polynomials with Higher Level
Authors: Komatsu Takao
Published: August 2021

There are many kinds of generalizations of Cauchy numbers and polynomials. Recently, a parametric type of the Bernoulli numbers with level 3 was introduced and studied as a kind of generalization of Bernoulli polynomials. A parametric type of Cauchy numbers with level 3 is its analogue. In this paper, as an analogue of a parametric type of Bernoulli polynomials with level 3 and its extension, we introduce a parametric type of Cauchy polynomials with a higher level. We present their characteristic and combinatorial properties. By using recursions, we show some determinant expressions.
Spatiotemporal Regulation and Functional Analysis of Circular RNAs in Skeletal Muscle and Subcutaneous Fat during Pig Growth
Authors: Li Biao , Yang Jinzeng , He Jun , Gong Yan , Xiao Yu , Zeng Qinghua , Xu Kang , Duan Yehui , He Jianhua , Ma Haiming
Published: August 2021

Recently, thousands of circular RNAs have been reported in different pig breeds. However, researches on the temporal and spatial expression patterns of circRNA over the period of animal growth are limited. Here, we systematically analyzed circRNAs in skeletal muscle and subcutaneous fat in four growth time points (30 days, 90 days, 150 days and 210 days after birth) of a Chinese native pig breed, Ningxiang pigs. A total of 1171 differentially expressed (DE) circRNAs between muscle and fat were identified, including 562 upregulated and 609 downregulated circRNAs. KEGG pathway enrichment analysis of these DE circRNAs revealed that host genes were mainly involved in glycogen metabolism signaling pathways, muscle development signaling pathways such as ErbB pathway and adipocytokine signaling pathways and AMPK signaling pathways and fatty acid biosynthesis. The circRNAs have striking spatiotemporal specificity in the form of dynamic expression at 90 d. Short Time-Series Expression Miner analysis showed multiple model spectra that are significantly enriched with time changes in muscle and fat. Our findings provide new ideas and perspectives about the role of circular RNAs and their targeting relations with mRNA and miRNA in skeletal muscle and fat tissue during pig growth.
Circadian Clock Gene Period Contributes to Diapause via GABAeric-Diapause Hormone Pathway in Bombyx mori
Authors: Cui Wen-Zhao , Qiu Jian-Feng , Dai Tai-Ming , Chen Zhuo , Li Jiang-Lan , Liu Kai , Wang Yu-Jun , Sima Yang-Hu , Xu Shi-Qing
Published: August 2021

Diapause is a developmental transition in insects based on seasonal adaptation to adversity; it is regulated by a circadian clock system and the endocrine system. However, the molecular node and its mechanism underlying the effects of these systems are still unclear. Here, a mutant of Bombyx mori with the circadian clock gene Period (Per) knocked out was constructed, which dramatically changed the classic diapause-destined pathway. Per-knockout silkworms powerfully attenuated, but could not completely block, the predetermined effects of temperature and photoperiod on diapause determination, and this effect depended on the diapause hormone (DH) pathway. The impaired transcription-translation feedback loop of the circadian clock system lacking the Per gene caused direct up-regulation of the expression of GRD, a receptor of γ-aminobutyric acid (GABA), by changing expression level of Cycle. The synthesis of GABA in the tissue complex of brain-suboesophageal ganglion then increased and restricted the decomposition, which continuously promoted the GABAergic signal to play a role, and finally inhibiting (delaying) the release of DH to the hemolymph, and reducing the diapause-inducing effect of DH. The results provided an example to explain the regulatory mechanism of the circadian clock on endocrine hormones in the silkworm.
Repetitive Sequence Distribution on Saguinus, Leontocebus and Leontopithecus Tamarins (Platyrrhine, Primates) by Mapping Telomeric (TTAGGG) Motifs and rDNA Loci
Authors: Ceraulo Simona , Perelman Polina L. , Mazzoleni Sofia , Rovatsos Michail , Dumas Francesca
Published: August 2021

Tamarins are a distinct group of small sized New World monkeys with complex phylogenetic relationships and poorly studied cytogenetic traits. In this study, we applied molecular cytogenetic analyses by fluorescence in situ hybridization with probes specific for telomeric sequences and ribosomal DNA loci after DAPI/CMA3 staining on metaphases from five tamarin species, namely Leontocebus fuscicollis, Leontopithecus rosalia, Saguinus geoffroyi, Saguinus mystax and Saguinus oedipus, with the aim to investigate the distribution of repetitive sequences and their possible role in genome evolution. Our analyses revealed that all five examined species show similar karyotypes, 2n = 46, which differ mainly in the morphology of chromosome pairs 16–17 and 19–22, due to the diverse distribution of rDNA loci, the amplification of telomeric-like sequences, the presence of heterochromatic blocks and/or putative chromosomal rearrangements, such as inversions. The differences in cytogenetic traits between species of tamarins are discussed in a comparative phylogenetic framework, and in addition to data from previous studies, we underline synapomorphies and apomorphisms that appeared during the diversification of this group of New World monkeys.
Lipocalin 2 Deficiency Restrains Aging-Related Reshaping of Gut Microbiota Structure and Metabolism
Authors: Qiu Xiaoxue , Chen Chi , Chen Xiaoli
Published: August 2021

Gut microbiota modulate age-associated changes in metabolism, innate immune responses, and cognitive function. However, the involvement of host factors in the regulation of age-dependent gut microbial structure and intestinal inflammation is largely unknown. Lipocalin 2 (Lcn2) has previously been identified as an adipocytokine and characterized as an important regulator of diet-induced obesity and inflammation. Previous studies have shown that Lcn2 plays a role in high fat diet-induced reshaping of gut microbiota and intestinal inflammation. However, the role of Lcn2 in the regulation of aging-related reshaping of gut microbiota is unclear. Herein, we demonstrate that fecal levels of Lcn2 are reduced during aging. Age reshaped gut microbiota composition in wild-type (WT) mice. Interestingly, Lcn2 deficiency diminished this effect of aging in Lcn2 knockout (LKO) mice, leading to decreased bacterial diversity and increased Firmicutes to Bacteroidetes (F to B) ratio. Specifically, we identified 16 bacteria at the family level that were differentially abundant between WT and LKO mice at old age. Several health-promoting bacteria, including SCFA-producing bacteria, were significantly less prevalent in old LKO mice compared to WT mice, indicating that Lcn2 deficiency shifts the aging-related gut microbial community towards an unhealthy population and lowers microbial butyrate production. Our results provide a line of evidence that Lcn2 plays a role in the control of aging-related reshaping of gut microbiota composition and metabolites.
Gold Nanoparticles: Multifaceted Roles in the Management of Autoimmune Disorders
Authors: Koushki Khadijeh , Keshavarz Shahbaz Sanaz , Keshavarz Mohsen , Bezsonov Evgeny E. , Sathyapalan Thozhukat , Sahebkar Amirhossein
Published: August 2021

Gold nanoparticles (GNPs) have been recently applied for various diagnostic and therapeutic purposes. The unique properties of these nanoparticles (NPs), such as relative ease of synthesis in various sizes, shapes and charges, stability, high drug-loading capacity and relative availability for modification accompanied by non-cytotoxicity and biocompatibility, make them an ideal field of research in bio-nanotechnology. Moreover, their potential to alleviate various inflammatory factors, nitrite species, and reactive oxygen production and the capacity to deliver therapeutic agents has attracted attention for further studies in inflammatory and autoimmune disorders. Furthermore, the characteristics of GNPs and surface modification can modulate their toxicity, biodistribution, biocompatibility, and effects. This review discusses in vitro and in vivo effects of GNPs and their functionalized forms in managing various autoimmune disorders (Ads) such as rheumatoid arthritis, type 1 diabetes, and multiple sclerosis.
Topological and Structural Plasticity of the Single Ig Fold and the Double Ig Fold Present in CD19
Authors: Youkharibache Philippe
Published: August 2021

The Ig fold has had a remarkable success in vertebrate evolution, with a presence in over 2% of human genes. The Ig fold is not just the elementary structural domain of antibodies and TCRs, it is also at the heart of a staggering 30% of immunologic cell surface receptors, making it a major orchestrator of cell–cell interactions. While BCRs, TCRs, and numerous Ig-based cell surface receptors form homo- or heterodimers on the same cell surface (in cis), many of them interface as ligand-receptors (checkpoints) on interacting cells (in trans) through their Ig domains. New Ig-Ig interfaces are still being discovered between Ig-based cell surface receptors, even in well-known families such as B7. What is largely ignored, however, is that the Ig fold itself is pseudosymmetric, a property that makes the Ig domain a versatile self-associative 3D structure and may, in part, explain its success in evolution, especially through its ability to bind in cis or in trans in the context of cell surface receptor–ligand interactions. In this paper, we review the Ig domains’ tertiary and quaternary pseudosymmetries, with particular attention to the newly identified double Ig fold in the solved CD19 molecular structure to highlight the underlying fundamental folding elements of Ig domains, i.e., Ig protodomains. This pseudosymmetric property of Ig domains gives us a decoding frame of reference to understand the fold, relate all Ig domain forms, single or double, and suggest new protein engineering avenues.
Wilson’s Disease: Facing the Challenge of Diagnosing a Rare Disease
Authors: Sánchez-Monteagudo Ana , Ripollés Edna , Berenguer Marina , Espinós Carmen
Published: August 2021

Wilson disease (WD) is a rare disorder caused by mutations in ATP7B, which leads to the defective biliary excretion of copper. The subsequent gradual accumulation of copper in different organs produces an extremely variable clinical picture, which comprises hepatic, neurological psychiatric, ophthalmological, and other disturbances. WD has a specific treatment, so that early diagnosis is crucial to avoid disease progression and its devastating consequences. The clinical diagnosis is based on the Leipzig score, which considers clinical, histological, biochemical, and genetic data. However, even patients with an initial WD diagnosis based on a high Leipzig score may harbor other conditions that mimic the WD’s phenotype (Wilson-like). Many patients are diagnosed using current available methods, but others remain in an uncertain area because of bordering ceruloplasmin levels, inconclusive genetic findings and unclear phenotypes. Currently, the available biomarkers for WD are ceruloplasmin and copper in the liver or in 24 h urine, but they are not solid enough. Therefore, the characterization of biomarkers that allow us to anticipate the evolution of the disease and the monitoring of new drugs is essential to improve its diagnosis and prognosis.
Salubrinal Enhances Cancer Cell Death during Glucose Deprivation through the Upregulation of xCT and Mitochondrial Oxidative Stress
Authors: Chen Mei-Chun , Hsu Li-Lin , Wang Sheng-Fan , Pan Yi-Ling , Lo Jeng-Fan , Yeh Tien-Shun , Tseng Ling-Ming , Lee Hsin-Chen
Published: August 2021

Cancer cells have the metabolic flexibility to adapt to heterogeneous tumor microenvironments. The integrated stress response (ISR) regulates the cellular adaptation response during nutrient stress. However, the issue of how the ISR regulates metabolic flexibility is still poorly understood. In this study, we activated the ISR using salubrinal in cancer cells and found that salubrinal repressed cell growth, colony formation, and migration but did not induce cell death in a glucose-containing condition. Under a glucose-deprivation condition, salubrinal induced cell death and increased the levels of mitochondrial reactive oxygen species (ROS). We found that these effects of salubrinal and glucose deprivation were associated with the upregulation of xCT (SLC7A11), which functions as an antiporter of cystine and glutamate and maintains the level of glutathione to maintain redox homeostasis. The upregulation of xCT did not protect cells from oxidative stress-mediated cell death but promoted it during glucose deprivation. In addition, the supplementation of ROS scavenger N-acetylcysteine and the maintenance of intracellular levels of amino acids via sulfasalazine (xCT inhibitor) or dimethyl-α-ketoglutarate decreased the levels of mitochondrial ROS and protected cells from death. Our results suggested that salubrinal enhances cancer cell death during glucose deprivation through the upregulation of xCT and mitochondrial oxidative stress.
Targeting ErbB3 Receptor in Cancer with Inhibitory Antibodies from Llama
Authors: Eliseev Igor E. , Ukrainskaya Valeria M. , Yudenko Anna N. , Mikushina Anna D. , Shmakov Stanislav V. , Afremova Anastasiya I. , Ekimova Viktoria M. , Vronskaia Anna A. , Knyazev Nickolay A. , Shamova Olga V.
Published: August 2021

The human ErbB3 receptor confers resistance to the pharmacological inhibition of EGFR and HER2 receptor tyrosine kinases in cancer, which makes it an important therapeutic target. Several anti-ErbB3 monoclonal antibodies that are currently being developed are all classical immunoglobulins. We took a different approach and discovered a group of novel heavy-chain antibodies targeting the extracellular domain of ErbB3 via a phage display of an antibody library from immunized llamas. We first produced three selected single-domain antibodies, named BCD090-P1, BCD090-M2, and BCD090-M456, in E. coli, as SUMO fusions that yielded up to 180 mg of recombinant protein per liter of culture. Then, we studied folding, aggregation, and disulfide bond formation, and showed their ultimate stability with half-denaturation of the strongest candidate, BCD090-P1, occurring in 8 M of urea. In surface plasmon resonance experiments, two most potent antibodies, BCD090-P1 and BCD090-M2, bound the extracellular domain of ErbB3 with 1.6 nM and 15 nM affinities for the monovalent interaction, respectively. The receptor binding was demonstrated by immunofluorescent confocal microscopy on four different ErbB3+ cancer cell lines. We observed that BCD090-P1 and BCD090-M2 bind noncompetitively to two distinct epitopes on the receptor. Both antibodies inhibited the ErbB3-driven proliferation of MCF-7 breast adenocarcinoma cells and HER2-overexpressing SK-BR-3 cells, with the EC50 in the range of 0.1–25 μg/mL. BCD090-M2 directly blocks ligand binding, whereas BCD090-P1 does not compete with the ligand and presumably acts through a distinct allosteric mechanism. We anticipate that these llama antibodies can be used to engineer new biparatopic anti-ErbB3 or bispecific anti-ErbB2/3 antibodies.
Hyaluronic Acid Hydrogels Crosslinked in Physiological Conditions: Synthesis and Biomedical Applications
Authors: Pérez Luis Andrés , Hernández Rebeca , Alonso José María , Pérez-González Raúl , Sáez-Martínez Virginia
Published: August 2021

Hyaluronic acid (HA) hydrogels display a wide variety of biomedical applications ranging from tissue engineering to drug vehiculization and controlled release. To date, most of the commercially available hyaluronic acid hydrogel formulations are produced under conditions that are not compatible with physiological ones. This review compiles the currently used approaches for the development of hyaluronic acid hydrogels under physiological/mild conditions. These methods include dynamic covalent processes such as boronic ester and Schiff-base formation and click chemistry mediated reactions such as thiol chemistry processes, azide-alkyne, or Diels Alder cycloaddition. Thermoreversible gelation of HA hydrogels at physiological temperature is also discussed. Finally, the most outstanding biomedical applications are indicated for each of the HA hydrogel generation approaches.
Super-Resolution Imaging with Graphene
Authors: Jiang Xiaoxiao , Kong Lu , Ying Yu , Gu Qiongchan , Lv Jiangtao , Dai Zhigao , Si Guangyuan
Published: August 2021

Super-resolution optical imaging is a consistent research hotspot for promoting studies in nanotechnology and biotechnology due to its capability of overcoming the diffraction limit, which is an intrinsic obstacle in pursuing higher resolution for conventional microscopy techniques. In the past few decades, a great number of techniques in this research domain have been theoretically proposed and experimentally demonstrated. Graphene, a special two-dimensional material, has become the most meritorious candidate and attracted incredible attention in high-resolution imaging domain due to its distinctive properties. In this article, the working principle of graphene-assisted imaging devices is summarized, and recent advances of super-resolution optical imaging based on graphene are reviewed for both near-field and far-field applications.
Developmental Dyslexia, Reading Acquisition, and Statistical Learning: A Sceptic’s Guide
Authors: Schmalz Xenia , Treccani Barbara , Mulatti Claudio
Published: August 2021

Many theories have been put forward that propose that developmental dyslexia is caused by low-level neural, cognitive, or perceptual deficits. For example, statistical learning is a cognitive mechanism that allows the learner to detect a probabilistic pattern in a stream of stimuli and to generalise the knowledge of this pattern to similar stimuli. The link between statistical learning and reading ability is indirect, with intermediate skills, such as knowledge of frequently co-occurring letters, likely being causally dependent on statistical learning skills and, in turn, causing individual variation in reading ability. We discuss theoretical issues regarding what a link between statistical learning and reading ability actually means and review the evidence for such a deficit. We then describe and simulate the “noisy chain hypothesis”, where each intermediary link between a proposed cause and the end-state of reading ability reduces the correlation coefficient between the low-level deficit and the end-state outcome of reading. We draw the following conclusions: (1) Empirically, there is evidence for a correlation between statistical learning ability and reading ability, but there is no evidence to suggest that this relationship is causal, (2) theoretically, focussing on a complete causal chain between a distal cause and developmental dyslexia, rather than the two endpoints of the distal cause and reading ability only, is necessary for understanding the underlying processes, (3) statistically, the indirect nature of the link between statistical learning and reading ability means that the magnitude of the correlation is diluted by other influencing variables, yielding most studies to date underpowered, and (4) practically, it is unclear what can be gained from invoking the concept of statistical learning in teaching children to read.
Alcohol Dependence in Rats Is Associated with Global Changes in Gene Expression in the Central Amygdala
Authors: Kisby Brent R. , Farris Sean P. , McManus Michelle M. , Varodayan Florence P. , Roberto Marisa , Harris R. Adron , Ponomarev Igor
Published: August 2021

Alcohol dependence is associated with adverse consequences of alcohol (ethanol) use and is evident in most severe cases of alcohol use disorder (AUD). The central nucleus of the amygdala (CeA) plays a critical role in the development of alcohol dependence and escalation of alcohol consumption in dependent subjects. Molecular mechanisms underlying the CeA-driven behavioral changes are not well understood. Here, we examined the effects of alcohol on global gene expression in the CeA using a chronic intermittent ethanol (CIE) vapor model in rats and RNA sequencing (RNA-Seq). The CIE procedure resulted in robust changes in CeA gene expression during intoxication, as the number of differentially expressed genes (DEGs) was significantly greater than those expected by chance. Over-representation analysis of cell types, functional groups and molecular pathways revealed biological categories potentially important for the development of alcohol dependence in our model. Genes specific for astrocytes, myelinating oligodendrocytes, and endothelial cells were over-represented in the DEG category, suggesting that these cell types were particularly affected by the CIE procedure. The majority of the over-represented functional groups and molecular pathways were directly related to the functions of glial and endothelial cells, including extracellular matrix (ECM) organization, myelination, and the regulation of innate immune response. A coordinated regulation of several ECM metalloproteinases (e.g., Mmp2; Mmp14), their substrates (e.g., multiple collagen genes and myelin basic protein; Mbp), and a metalloproteinase inhibitor, Reck, suggests a specific mechanism for ECM re-organization in response to chronic alcohol, which may modulate neuronal activity and result in behavioral changes, such as an escalation of alcohol drinking. Our results highlight the importance of glial and endothelial cells in the effects of chronic alcohol exposure on the CeA, and demonstrate further insight into the molecular mechanisms of alcohol dependence in rats. These molecular targets may be used in future studies to develop therapeutics to treat AUD.
Histological Grade of Endometrioid Endometrial Cancer and Relapse Risk Can Be Predicted with Machine Learning from Gene Expression Data
Authors: Gargya Péter , Bálint Bálint László
Published: August 2021

The tumor grade of endometrioid endometrial cancer is used as an independent marker of prognosis and a key component in clinical decision making. It is reported that between grades 1 and 3, however, the intermediate grade 2 carries limited information; thus, patients with grade 2 tumors are at risk of both under- and overtreatment. We used RNA-sequencing data from the TCGA project and machine learning to develop a model which can correctly classify grade 1 and grade 3 samples. We used the trained model on grade 2 patients to subdivide them into low-risk and high-risk groups. With iterative retraining, we selected the most relevant 12 transcripts to build a simplified model without losing accuracy. Both models had a high AUC of 0.93. In both cases, there was a significant difference in the relapse-free survivals of the newly identified grade 2 subgroups. Both models could identify grade 2 patients that have a higher risk of relapse. Our approach overcomes the subjective components of the histological evaluation. The developed method can be automated to perform a prescreening of the samples before a final decision is made by pathologists. Our translational approach based on machine learning methods could allow for better therapeutic planning for grade 2 endometrial cancer patients.
A Nuclear-Directed Ribonuclease Variant Targets Cancer Stem Cells and Inhibits Migration and Invasion of Breast Cancer Cells
Authors: Castro Jessica , Tornillo Giusy , Ceada Gerardo , Ramos-Neble Beatriz , Bravo Marlon , Ribó Marc , Vilanova Maria , Smalley Matthew J. , Benito Antoni
Published: August 2021

Despite the significant advances in cancer research made in recent years, this disease remains one of the leading causes of death worldwide. In part, this is due to the fact that after therapy, a subpopulation of self-renewing tumor cells can survive and promote cancer relapse, resistance to therapies and metastasis. Targeting these cancer stem cells (CSCs) is therefore essential to improve the clinical outcome of cancer patients. In this sense, multi-targeted drugs may be promising agents targeting CSC-associated multifocal effects. We have previously constructed different human pancreatic ribonuclease (RNase) variants that are cytotoxic for tumor cells due to a non-classical nuclear localization signal introduced in their sequence. These cytotoxic RNases affect the expression of multiple genes involved in deregulated metabolic and signaling pathways in cancer cells and are highly cytotoxic for multidrug-resistant tumor cell lines. Here, we show that these cytotoxic nuclear-directed RNases are highly selective for tumor cell lines grown in 3D, inhibit CSCs’ development and diminish the self-renewal capacity of the CSCs population. Moreover, these human RNase variants reduce the migration and invasiveness of highly invasive breast cancer cells and downregulate N-cadherin expression.
Minimally Invasive Approach to Gastric GISTs: Analysis of a Multicenter Robotic and Laparoscopic Experience with Literature Review
Authors: Ceccarelli Graziano , Costa Gianluca , De Rosa Michele , Codacci Pisanelli Massimo , Frezza Barbara , De Prizio Marco , Bravi Ilaria , Scacchi Andrea , Gallo Gaetano , Amato Bruno , Bugiantella Walter , Tacchi Piergiorgio , Bartoli Alberto , Patriti Alberto , Cappuccio Micaela , Komici Klara , Mariani Lorenzo , Avella Pasquale , Rocca Aldo
Published: August 2021

Background: Gastrointestinal stromal tumors (GISTs) are most frequently located in the stomach. In the setting of a multidisciplinary approach, surgery represents the best therapeutic option, consisting mainly in a wedge gastric resection. (1) Materials and methods: Between January 2010 to September 2020, 105 patients with a primary gastrointestinal stromal tumor (GISTs) located in the stomach, underwent surgery at three surgical units. (2) Results: A multi-institutional analysis of minimally invasive series including 81 cases (36 laparoscopic and 45 robotic) from 3 referral centers was performed. Males were 35 (43.2%), the average age was 66.64 years old. ASA score ≥3 was 6 (13.3%) in the RS and 4 (11.1%) in the LS and the average tumor size was 4.4 cm. Most of the procedures were wedge resections (N = 76; 93.8%) and the main operative time was 151 min in the RS and 97 min in the LS. Conversion was necessary in five cases (6.2%). (3) Conclusions: Minimal invasive approaches for gastric GISTs performed in selected patients and experienced centers are safe. A robotic approach represents a useful option, especially for GISTs that are more than 5 cm, even located in unfavorable places.
Photodynamic Therapy for Pancreatic Ductal Adenocarcinoma
Authors: Karimnia Vida , Slack Frank J. , Celli Jonathan P.
Published: August 2021

Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal of human cancers. Clinical trials of various chemotherapy, radiotherapy, targeted agents and combination strategies have generally failed to provide meaningful improvement in survival for patients with unresectable disease. Photodynamic therapy (PDT) is a photochemistry-based approach that enables selective cell killing using tumor-localizing agents activated by visible or near-infrared light. In recent years, clinical studies have demonstrated the technical feasibility of PDT for patients with locally advanced PDAC while a growing body of preclinical literature has shown that PDT can overcome drug resistance and target problematic and aggressive disease. Emerging evidence also suggests the ability of PDT to target PDAC stroma, which is known to act as both a barrier to drug delivery and a tumor-promoting signaling partner. Here, we review the literature which indicates an emergent role of PDT in clinical management of PDAC, including the potential for combination with other targeted agents and RNA medicine.
The Molecular Networks of microRNAs and Their Targets in the Drug Resistance of Colon Carcinoma
Authors: Crudele Francesca , Bianchi Nicoletta , Astolfi Annalisa , Grassilli Silvia , Brugnoli Federica , Terrazzan Anna , Bertagnolo Valeria , Negrini Massimo , Frassoldati Antonio , Volinia Stefano
Published: August 2021

Drug resistance is one of the major forces driving a poor prognosis during the treatment and progression of human colon carcinomas. The molecular mechanisms that regulate the diverse processes underlying drug resistance are still under debate. MicroRNAs (miRNAs) are a subgroup of non-coding RNAs increasingly found to be associated with the regulation of tumorigenesis and drug resistance. We performed a systematic review of the articles concerning miRNAs and drug resistance in human colon cancer published from 2013 onwards in journals with an impact factor of 5 or higher. First, we built a network with the most studied miRNAs and targets (as nodes) while the drug resistance/s are indicated by the connections (edges); then, we discussed the most relevant miRNA/targets interactions regulated by drugs according to the network topology and statistics. Finally, we considered the drugs as nodes in the network, to allow an alternative point of view that could flow through the treatment options and the associated molecular pathways. A small number of microRNAs and proteins appeared as critically involved in the most common drugs used for the treatment of patients with colon cancer. In particular, the family of miR-200, miR34a, miR-155 and miR-17 appear as the most relevant microRNAs. Thus, regulating these miRNAs could be useful for interfering with some drug resistance mechanisms in colorectal carcinoma.