bluuebunny/crossref_metadata_2025 huggingface dataset:
Applications of the Neutrosophic Poisson Distribution for Bi-Univalent Functions Involving the Modified Caputo’s Derivative Operator
Authors: Santhiya S. , Thilagavathi K.
Published: December 2022

This paper establishes the upper bounds for the second and third coefficients of holomorphic and bi-univalent functions in a family which involves Bazilevic functions and μ-pseudo-starlike functions under a new operator, joining the neutrosophic Poisson distribution with the modified Caputo’s derivative operator. We also discuss Fekete–Szego’s function problem in this family. Examples are given to support our case for the neutrosophic Poisson distribution. The fields of physics, mechanics, engineering, and biology all make extensive use of fractional derivatives.
Intervertebral Disc Tissue Engineering Using Additive Manufacturing
Authors: Yoshida Minami , Turner Paul Richard , Cabral Jaydee Dones
Published: December 2022

Intervertebral disc (IVD) degeneration is one of the major causes of lower back pain, a common health condition that greatly affects the quality of life. With an increasing elderly population and changes in lifestyle, there exists a high demand for novel treatment strategies for damaged IVDs. Researchers have investigated IVD tissue engineering (TE) as a way to restore biological and mechanical functions by regenerating or replacing damaged discs using scaffolds with suitable cells. These scaffolds can be constructed using material extrusion additive manufacturing (AM), a technique used to build three-dimensional (3D), custom discs utilising computer-aided design (CAD). Structural geometry can be controlled via the manipulation of printing parameters, material selection, temperature, and various other processing parameters. To date, there are no clinically relevant TE-IVDs available. In this review, advances in AM-based approaches for IVD TE are briefly discussed in order to achieve a better understanding of the requirements needed to obtain more effective, and ultimately clinically relevant, IVD TE constructs.
Integrative Analysis of miRNAs Involved in Fat Deposition in Different Pig Breeds
Authors: Zhang Xiuxiu , Huang Wanlong , Guo Yuntao , Miao Xiangyang
Published: December 2022

Background: miRNAs are a set of small, noncoding RNAs that bind to partially complementary sequences on target mRNAs. This leads to the post-transcriptional regulation of gene expression. Many studies have shown that microRNAs play critical roles in adipose cell differentiation and fat metabolism. The aim of this study was to explore the regulatory functions of miRNAs in fat deposition for the prevention and therapy of lipid metabolism-related diseases. Methods: The significant differences in the fat deposition of Laiwu (LW) pigs and Large White (LY) pigs were studied. To investigate the genetic relationships of miRNAs that regulate fat deposition, we performed a genome-wide analysis of miRNAs derived from subcutaneous adipose tissue of LW and LY pigs using RNA-seq. Results: There were 39 known miRNAs and 56 novel miRNAs significantly differential expressed between the two breeds of pigs. In the analysis of the Gene Ontology and KEGG pathways, predicted targets of these differentially expressed miRNAs were involved in several fat-associated pathways, such as the peroxisome proliferator-activated receptor (PPAR), mitogen-activated protein kinases (MAPK) and Wnt signaling pathways. In addition, ssc-miR-133a-3p, ssc-miR-486 and ssc-miR-1 each had a great impact on the development of porcine subcutaneous fat through the PPAR signaling pathway. Conclusions: We explored the role of differentially expressed miRNAs and studied the mechanisms of adipogenesis and fat deposition between two different pig breeds. In addition, these results also contribute to research relevant to human obesity.
Genome-Wide Identification, Characterization, and Expression Analysis of GRAS Gene Family in Ginger (Zingiber officinale Roscoe)
Authors: Tian Shuming , Wan Yuepeng , Jiang Dongzhu , Gong Min , Lin Junyao , Xia Maoqin , Shi Cuiping , Xing Haitao , Li Hong-Lei
Published: December 2022

GRAS family proteins are one of the most abundant transcription factors in plants; they play crucial roles in plant development, metabolism, and biotic- and abiotic-stress responses. The GRAS family has been identified and functionally characterized in some plant species. However, this family in ginger (Zingiber officinale Roscoe), a medicinal crop and non-prescription drug, remains unknown to date. In the present study, 66 GRAS genes were identified by searching the complete genome sequence of ginger. The GRAS family is divided into nine subfamilies based on the phylogenetic analyses. The GRAS genes are distributed unevenly across 11 chromosomes. By analyzing the gene structure and motif distribution of GRAS members in ginger, we found that the GRAS genes have more than one cis-acting element. Chromosomal location and duplication analysis indicated that whole-genome duplication, tandem duplication, and segmental duplication may be responsible for the expansion of the GRAS family in ginger. The expression levels of GRAS family genes are different in ginger roots and stems, indicating that these genes may have an impact on ginger development. In addition, the GRAS genes in ginger showed extensive expression patterns under different abiotic stresses, suggesting that they may play important roles in the stress response. Our study provides a comprehensive analysis of GRAS members in ginger for the first time, which will help to better explore the function of GRAS genes in the regulation of tissue development and response to stress in ginger.
Analysis of SNHG14: A Long Non-Coding RNA Hosting SNORD116, Whose Loss Contributes to Prader–Willi Syndrome Etiology
Authors: Ariyanfar Shadi , Good Deborah J.
Published: December 2022

The Small Nucleolar Host Gene 14 (SNHG14) is a host gene for small non-coding RNAs, including the SNORD116 small nucleolar C/D box RNA encoding locus. Large deletions of the SNHG14 locus, as well as microdeletions of the SNORD116 locus, lead to the neurodevelopmental genetic disorder Prader–Willi syndrome. This review will focus on the SNHG14 gene, its expression patterns, its role in human cancer, and the possibility that single nucleotide variants within the locus contribute to human phenotypes in the general population. This review will also include new in silico data analyses of the SNHG14 locus and new in situ RNA expression patterns of the Snhg14 RNA in mouse midbrain and hindbrain regions.
A Multi-proxy Provenance Study of Late Carboniferous to Middle Jurassic Sandstones in the Eastern Sverdrup Basin and Its Bearing on Arctic Palaeogeographic Reconstructions
Authors: Pointon Michael A. , Smyth Helen , Omma Jenny E. , Morton Andrew C. , Schneider Simon , Hülse Peter , Rippington Stephen J. , Lopez-Mir Berta , Crowley Quentin G. , Millar Ian , Whitehouse Martin J. , Frei Dirk , Scott Robert A. , Flowerdew Michael J.
Published: December 2022

A multi-proxy provenance study of Late Carboniferous to Middle Jurassic sandstones from the eastern Sverdrup Basin was undertaken employing optical petrography and heavy mineral analysis, chemical analysis of apatite, garnet and rutile grains, as well as detrital zircon U–Pb geochronology and Hf isotope analysis. Late Carboniferous to Middle Jurassic strata on the southern basin margin are inferred as being predominantly reworked from Silurian to Devonian strata within the adjacent Franklinian Basin succession. Higher-grade metamorphic detritus appeared during Middle to Late Triassic times and indicates exhumation and erosion of lower (Neoproterozoic to Cambrian) levels within the Franklinian Basin succession and/or a direct detrital input from the Canadian-Greenland Shield. The provenance of northern-derived sediments is more enigmatic owing to the subsequent opening of the Arctic Ocean. Northern-derived Middle Permian to Early Triassic sediments were likely derived from proximal areas of the Chukotkan part of the Arctic Alaska-Chukotka microplate. Late Triassic northern-derived sediments have different detrital zircon U–Pb age spectra from Middle Permian to Early Triassic ones and were likely derived from the Uralian orogenic belt and/or the Arctic Uralides. The loss of this sand input during latest Triassic times is interpreted to reflect drainage reorganisation farther upstream on the Barents Shelf. Middle Jurassic sands in the northern and axial parts of the basin were largely reworked from local northern-derived Late Triassic strata. This may have been facilitated by rift flank uplift of the northern basin margin in response to rifting in the adjacent proto-Amerasia Basin.
Application Software Developed for the Determination of Expansion Volume in Clay Soil Generated by the Detonation of an Explosive Charge
Authors: Težak Denis , Dobrilović Ivana , Dobrilović Mario , Jurenić Davorin
Published: December 2022

Blasting in clay soil in the field of anchoring and foundation of objects and structures has its benefits in construction and geotechnical practice. The foundations of the method lay in the fact that a shock wave is generated when the explosive charge is detonated. The shock wave, with high pressure at the wavefront, causes the natural structure of the clay soil to be destroyed, and a spherical expansion in the clay mass is formed. The presented research is focused on determining the shape and volume of the resulting expansion in test blasts performed with several types of explosives. An application named Borehole was developed to determine the resulting spherical expansion formed after the detonation of an explosive charge with the integration of the GNSS method of measurement, depth camera, and laser. The application Borehole calculates expansion volume based on the coordinates obtained with the GNSS and the laser-obtained distance of the formed expansion and provides a graphical interpretation in 2D and 3D views. Additionally, when developing the application Borehole, compatibility with CAD tools was considered, primarily for better verification and a more detailed graphical interpretation of 3D views. The developed method allows for simple determination of the volume and dimension of the spherical expansion in clay soil with acceptable accuracy for the design and building of geotechnical structures constructed above and underground.
Self-Reported Reasons for Inconsistent Participation in Colorectal Cancer Screening Using FIT in Flanders, Belgium
Authors: Hoeck Sarah , Tran Thuy Ngan
Published: December 2022

Background: In Flanders, the uptake in the population-based colorectal cancer (CRC) screening program (using fecal immunochemical test, FIT) is suboptimal (~50%). This study explored the reasons for inconsistent participation in FIT screening among irregular participants in Flanders. Methods: An online survey with both open questions and fixed statements was sent to irregular participants (2016–2018) in the Flemish CRC screening program. A reminder email followed eight weeks after the first email. Data analysis used both qualitative and quantitative approaches. Post-stratification weights based on gender, age group, and the first two digits of the postcode were employed to reduce non-response bias. Results: In total, 5328 out of 19,592 irregular participants responded to the survey. While the main reasons not to participate were related to ‘postponing participation’ and ‘having other priorities’, the main reasons to participate were related to the importance of (preventive) health checks. The role of general practitioners (GPs) in promoting CRC screening also emerged as an important theme among the respondents’ answers (based on fixed statements). Conclusions: The study reported the main reasons for inconsistent participation in FIT screening for CRC in Flanders. The findings are helpful in guiding tailored interventions to increase FIT screening uptake in the region.
What Is Known about Midazolam? A Bibliometric Approach of the Literature
Authors: Corôa Maria Claudia Pinheiro , Mendes Paulo Fernando Santos , Baia-da-Silva Daiane Claydes , Souza-Monteiro Deiweson , Ferreira Maria Karolina Martins , Braga Glenda Luciana Costa , Damasceno Taissa Viana , Perdigão José Messias , Lima Rafael Rodrigues
Published: December 2022

Midazolam is a drug with actions towards the central nervous system producing sedative and anticonvulsants effects, used for sedation and seizures treatments. A better understanding about its effects in the different scenarios presented in the literature could be helpful to gather information regarding its clinical indications, pharmacological interactions, and adverse events. From this perspective, the aim of this study was to analyze the global research about midazolam mapping, specifically the knowledge of the 100 most-cited papers about this research field. For this, a search was executed on the Web of Science-Core Collection database using bibliometric methodological tools. The search strategy retrieved 34,799 articles. A total of 170 articles were evaluated, with 70 articles being excluded for not meeting the inclusion criteria. The 100 most-cited articles rendered 42,480 citations on WoS-CC, ranging from 253 to 1744. Non-systematic review was the most published study type, mainly from North America, during the period of 1992 to 2002. The most frequent keywords were midazolam and pharmacokinetics. Regarding the authors, Thummel and Kunze were the ones with the greatest number of papers included. Our findings showed the global research trends about midazolam, mainly related to its different effects and uses throughout the time.
Assistive Technologies and Quadriplegia: A Map Point on the Development and Spread of the Tongue Barbell Piercing
Authors: Pirrera Antonia , Meli Paola , De Dominicis Alessandra , Lepri Alessandra , Giansanti Daniele
Published: December 2022

The barbell piercing can be used as an assistive device that allows people with severe disabilities, such as tetraplegia, to control their environments using the movement of the tongue. The human tongue can move rapidly and accurately, such that the tip can touch every tooth. Lingual control systems allow people with disabilities to take advantage of their residual skills for easier communication and to improve the control of mobility and the surrounding environment. The aim of this study was to conduct a narrative review of the development and dissemination of the assistive technologies based on tongue control by means of the barbell piercing. The design of the study was based on: (I) an overview of Pubmed complemented with other databases and Web searches (also institutional); (II) an organization according to a standardized checklist for narrative reviews; (III) an arrangement with four different perspectives: the trends in the scientific literature, technological evolution and categorization, dominant approaches, issues of incorporation into the health domain—such as acceptance, safety, and regulations. The results have highlighted: (1) that the volume of scientific productions, which started in this sector before the smartphone expansion, has not increased; (2) that it is possible to make a map point of the technological evolution and categorization; (3) that these assistive technologies have a high degree of acceptance and performance, especially when integrated with aid tools with mechatronics; (4) and the complexity of the regulatory framework in this area. The study, from a general point of view, highlighted the high potential of these systems and we suggest investing the energy into agreement tools for assistive technologies (AT)s, such as health technology assessment studies, comparative assessment analysis, or consensus conferences that could allow a better diffusion and use of ATs, including these systems.
Violence, Harassment, and Turnover Intention in Home and Community Care: The Role of Training
Authors: Sayin Firat K. , Denton Margaret , Brookman Catherine , Davies Sharon , Zeytinoglu Isik U.
Published: December 2022

Background: Violence and harassment affect healthcare workers’ well-being and career decisions in the home and community care sector. Purpose: The objective of this study is to assess the role of training in alleviating the relationship between violence and harassment at work and turnover intention among personal support workers (PSWs). Methodology/Approach: Cross-sectional survey data from 1401 PSWs in Ontario, Canada are analyzed with structural equation modeling. Utilizing a resource perspective, the associations between job demands (i.e., violence and harassment at work), personal resources (i.e., self-esteem), job resources (i.e., workplace violence training and challenging task training), stress, and intention to stay among personal support workers (PSWs) are examined. Results: Challenging task training is positively associated with self-esteem and negatively associated with stress, whereas workplace violence training does not have a significant association with either variable. Stress has a negative relationship with intention to stay. Self-esteem is the mediator of both associations between violence and harassment at work and stress and between challenging task training and stress. Discussion: The results point to varied degrees of training effectiveness that may be shaping turnover decisions of PSWs who experience violence and harassment in home and community care organizations. Practice implications: There seems to be a need to assess and redesign workplace violence training. Home and community care managers might be able to lower the impact of violence and harassment on PSWs’ turnover by providing training that is not directly related to workplace violence and harassment.
The Effect of Emotional Labor on the Physical and Mental Health of Health Professionals: Emotional Exhaustion Has a Mediating Effect
Authors: Chen Chien-Chih , Lan Yu-Li , Chiou Shau-Lun , Lin Yi-Ching
Published: December 2022

(1) Background: Workers who perform emotional labor for an extended period are prone to emotional exhaustion; in particular, when the work exceeds the range of one’s emotional resources, it will produce job burnout. This study investigated the effects of emotional labor and emotional exhaustion on the physical and mental health of health professionals. (2) Methods: This study was cross-sectional and the sampling criteria were health professionals from August 2020 to July 2021, including rehabilitators, nutritionists, clinical psychologists, radiologists, respiratory therapists, pharmacists, medical examiners and audiologists. A questionnaire was used to collect data on participants’ emotional labor, emotional exhaustion, physical health and mental health. A total of 120 valid questionnaires were obtained. (3) Results: Significant positive correlations were found between emotional labor and emotional exhaustion, physical and mental health and anxiety. A hierarchical regression analysis found that the effect of emotional labor on physical and mental health increased the predictive power to 59.7% through emotional exhaustion, and emotional exhaustion had a mediating effect on the relationship between emotional labor and physical and mental health. (4) Conclusions: This study provides a reference for managers of medical institutions to care for employees’ work stress and physical and mental health, which will help institutions build a friendly and healthy workplace.
Development of a Digital Healthcare Management System for Lower-Extremity Amputees: A Pilot Study
Authors: Kim Jin Hong , Kim Yu Ri , Han Mi Hyang , Lee Ji Young , Kim Ji Sung , Kang Yong Cheol , Yoon Seong Jun , Chang Yunhee , Lee Gangpyo , Cho Nam Soon
Published: December 2022

The research, which was designed as a “pre- and post-single group” study, included patients with lower-limb amputation and aimed to evaluate the effectiveness of self-directed physical-strength training and cardiovascular exercise using a novel digital healthcare management service three times a week for 12 weeks. Muscle strength, thigh circumference, lipid profile and glycated hemoglobin levels, pulmonary function, quality of life, and physical activity level were evaluated before and after the intervention, while satisfaction was measured after the study. Among the 14 included patients, the proportion of adherence to the physical-strength training and physical-strengthening activity were 85.2% and 75.8%, respectively. The level of satisfaction with the digital healthcare management system was high. Significant changes were observed in the muscle-strength tests (dominant grip power and muscle strength of knee flexion and extension of the intact side), thigh circumference, and glycated hemoglobin levels. Further, the quality-of-life score showed improvement, although without significant differences. Individualized exercise management using the novel digital healthcare management system for lower-limb amputees could induce interest in self-care and promote physical activity and healthy behavior. Through this effect, we can expect a reduction in the incidence of cardiovascular diseases, diabetes mellitus, dyslipidemia, and severe injuries from falling.
Zooarchaeological Evidence from Medieval Ojców Castle, Lesser Poland
Authors: Religa-Sobczyk Joanna , Wertz Krzysztof , Lõugas Lembi , Wojenka Michał , Lemanik Anna , Wojtal Piotr
Published: December 2022

Archaeological research at Ojców castle has yielded important information about life in that medieval castle. The results of zooarchaeological analyses presented in this paper complement the archaeological research, adding to our knowledge of the diet of the castle inhabitants from the time of establishment of the castle until the final residents. Zooarchaeological research is also complemented by data from older settlement phases on the castle hill, directly related to the Lusatian culture in the early Iron Age. The great variability of remains from mammals, birds, and fish and the taphonomic features of bones found in the different chronological strata of the castle’s courtyard reflect the diverse economic activities that took place in particular times and spaces.
Agrarian Archaeology: A Research and Social Transformation Tool
Authors: Fernández Mier Margarita , Fernández Fernández Jesús , López Gómez Pablo
Published: December 2022

The aim of this paper is to approach the concept of agrarian archaeology, an epistemological framework that allows the investigation of agrarian landscapes favouring new historical narratives far from traditional paradigms and, at the same time, the proposal of innovative forms of cultural heritage management in rural areas. The working methodology and some examples of both empirical work (archaeology) and research-action approaches to cultural heritage are presented. As a result, agrarian archaeology opens up a perspective of experimental research, which permits the problematization of conceptual languages, questions historical narratives and causes the rethinking of personal practices, in addition to promoting an open science close to citizens. Cultural heritage here is to be understood as a social process of meaningful construction, a space of debate and transformation for social innovation.
The Influence of Government Regulation on Farmers’ Green Production Behavior—From the Perspective of the Market Structure
Authors: Huang Qiang , Wang Huizhu , Chen Chao
Published: December 2022

To clarify the factors influencing the green production behavior of peach farmers, this paper uses the survey data of 741 peach farmers in 19 provinces and cities in China, it uses a multiple ordered probit model to empirically analyze the impact of the government regulations on the green production behavior of peach farmers, from the perspective of the market structure. This paper also analyzes its intermediary role in this process, and it analyzes the possible heterogeneity at both the planting scale and the regional level. The results show the following: (1) Government regulation has a positive and significant impact on the green production behavior of peach farmers. Specifically, government supervision and inspection, alongside green subsidies, can positively promote the implementation of green production behavior by peach farmers, but government publicity and training have not played a good role. (2) The market structure plays a partial intermediary role, rather than a complete intermediary role, in the government regulation affecting the green production behavior of peach farmers. (3) The impact of the government regulation on the green production behavior of peach farmers is heterogeneous. Specifically, compared with small farmers, the impact on large-scale farmers is higher; however, the influence of the three methods of government regulation on the green production behavior of peach farmers varies from region to region. Therefore, in order to promote the implementation of green production, the government should introduce appropriate local policies, strongly support new agricultural business entities, draw clear guidelines for the market, and play the role of “night watchman”.
Are COL22A1 Gene Polymorphisms rs11784270 and rs6577958 Associated with Susceptibility to a Non-Contact Anterior Cruciate Ligament Injury in Polish Athletes?
Authors: Sun Zhuo , Cięszczyk Paweł , Lulińska Ewelina , Dzitkowska-Zabielska Magdalena , Johne Monika , Humińska-Lisowska Kinga , Michałowska-Sawczyn Monika , Ficek Krzysztof , Leońska-Duniec Agata , Mastalerz Andrzej , Janczyk Arkadiusz , Marek Sawczuk
Published: December 2022

Understanding the risk factors and etiology of ACL ruptures (anterior cruciate ligament) is crucial due to the injury’s high occurrence, significant financial cost to the healthcare sector, and clinical consequences. In this study, we investigated the hypothesis that rs11784270 A/C and rs6577958 C/T SNPs (single gene polymorphism) within COL22A1 are associated with ACL ruptures (ACLR) in Polish soccer players. Methods: 228 athletes with ACLR (157 male, age 26 ± 4, 71 female, age 26 ± 6) and 202 control athletes (117 male, age 26 ± 6, 85 female, age 29 ± 2) engaged in the study. The buccal cell swabs were genotyped using TaqMan® pre-designed SNP genotyping assays, following the manufacturer’s recommendations. The R program and SNPassoc package were used to determine the genotype and allele frequency distributions under the various inheritance models (co-dominant, dominant, recessive, and over-dominant). Further, p-values of <0.05 were considered statistically significant. We found no association between the analyzed polymorphisms and the risk of non-contact ACL ruptures in any of the studied models. Although the genetic variants investigated in this study were not associated with the risk of non-contact ACL ruptures, we assumed that the COL22A1 gene remains a candidate for further investigations in musculoskeletal injuries.
Attentional Prioritization of Infant Faces in Parents: The Influence of Parents’ Experiences of Care
Authors: Gemignani Micol , Giannotti Michele , Schmalz Xenia , Rigo Paola , De Falco Simona
Published: December 2022

Infant faces are prioritized by the attentional system in parents, resulting in a greater cognitive engagement in terms of response time. However, many biological, contextual and environmental factors relating to this cognitive mechanism have been left unexplored. To fill this gap, this study aims to (i) confirm that infant faces engage more attention compared to adult faces; (ii) investigate whether the attention to infant faces is affected early care experiences of parents; (iii) explore the effect of parents’ sex by taking the amount of involvement with early childcare into consideration. 51 mothers and 46 fathers completed a modified Go/no-Go task, a brief sociodemographic questionnaire, the short version of the Adult Parental Acceptance–Rejection scale, and an ad-hoc question relating to the amount of parental involvement with early childcare. Parents’ response times were slowed in the presence of infant versus adult faces. Parents whose mother was perceived as more sensitively accepting were more engaged by infant cues. By considering the amount of early parental involvement, the sex of parents did not significantly interact with the type of face. These findings provide new insights on the attention process in response to infant cues in parents and suggest that the investigation of experience-based factors may shed further light on this topic.
Therapeutic Targeting of the Proteolytic Enzymes
Authors: Couture Frédéric
Published: December 2022

The processes regulating the generation of proteins from the early translation events to the final biologically active products are complex and tightly controlled [...]
Simultaneous Antagonism at H3R/D2R/D3R Reduces Autism-like Self-Grooming and Aggressive Behaviors by Mitigating MAPK Activation in Mice
Authors: Eissa Nermin , Awad Mohamed Al , Thomas Shilu Deepa , Venkatachalam Karthikkumar , Jayaprakash Petrilla , Zhong Sicheng , Stark Holger , Sadek Bassem
Published: December 2022

Dysregulation in brain neurotransmitters underlies several neuropsychiatric disorders, e.g., autism spectrum disorder (ASD). Also, abnormalities in the extracellular-signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) pathway pave the way for neuroinflammation, neurodegeneration, and altered learning phenotype in ASD. Therefore, the effects of chronic systemic administration of the multiple-targeting antagonist ST-713 at the histamine H3 receptor (H3R) and dopamine D2/D3 receptors (D2/D3R) on repetitive self-grooming, aggressive behaviors, and abnormalities in the MAPK pathway in BTBR T + Itpr3tf/J (BTBR) mice were assessed. The results showed that ST-713 (2.5, 5, and 10 mg/kg, i.p.) mitigated repetitive self-grooming and aggression in BTBR mice (all p < 0.05), and the ameliorative effects of the most promising dose of ST-713 (5 mg/kg, i.p.) on behaviors were completely abrogated by co-administration of the H3R agonist (R)-α-methylhistamine or the anticholinergic drug scopolamine. Moreover, the elevated levels of several MAPK pathway proteins and induced proinflammatory markers such as tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and IL-6 were significantly suppressed following chronic administration of ST-713 (5 mg/kg, i.p.) (all p < 0.01). Furthermore, ST-713 significantly increased the levels of histamine and dopamine in hippocampal tissue of treated BTBR mice (all p < 0.01). The current observations signify the potential role of such multiple-targeting compounds, e.g., ST-713, in multifactorial neurodevelopmental disorders such as ASD.
Electroactive Hydroxyapatite/Carbon Nanofiber Scaffolds for Osteogenic Differentiation of Human Adipose-Derived Stem Cells
Authors: Sun Baojun , Sun Yajie , Han Shuwei , Zhang Ruitong , Wang Xiujuan , Meng Chunxia , Ji Tuo , Sun Chunhui , Ren Na , Ge Shaohua , Liu Hong , Yu Yang , Wang Jingang
Published: December 2022

Traditional bone defect treatments are limited by an insufficient supply of autologous bone, the immune rejection of allogeneic bone grafts, and high medical costs. To address this medical need, bone tissue engineering has emerged as a promising option. Among the existing tissue engineering materials, the use of electroactive scaffolds has become a common strategy in bone repair. However, single-function electroactive scaffolds are not sufficient for scientific research or clinical application. On the other hand, multifunctional electroactive scaffolds are often complicated and expensive to prepare. Therefore, we propose a new tissue engineering strategy that optimizes the electrical properties and biocompatibility of carbon-based materials. Here, a hydroxyapatite/carbon nanofiber (HAp/CNF) scaffold with optimal electrical activity was prepared by electrospinning HAp nanoparticle-incorporated polyvinylidene fluoride (PVDF) and then carbonizing the fibers. Biochemical assessments of the markers of osteogenesis in human adipose-derived stem cells (h-ADSCs) cultured on HAp/CNF scaffolds demonstrate that the material promoted the osteogenic differentiation of h-ADSCs in the absence of an osteogenic factor. The results of this study show that electroactive carbon materials with a fibrous structure can promote the osteogenic differentiation of h-ADSCs, providing a new strategy for the preparation and application of carbon-based materials in bone tissue engineering.
B-Cell Epitope Mapping of the Vibrio cholera Toxins A, B, and P and an ELISA Assay
Authors: De-Simone Salvatore G. , Napoleão-Pêgo Paloma , Gonçalves Priscilla S. , Lechuga Guilherme C. , Cardoso Sergian V. , Provance David W. , Morel Carlos M. , da Silva Flavio R.
Published: December 2022

Oral immunization with the choleric toxin (CT) elicits a high level of protection against its enterotoxin activities and can control cholera in endemic settings. However, the complete B-cell epitope map of the CT that is responsible for protection remains to be clarified. A library of one-hundred, twenty-two 15-mer peptides covering the entire sequence of the three chains of the CT protein (CTP) was prepared by SPOT synthesis. The immunoreactivity of membrane-bound peptides with sera from mice vaccinated with an oral inactivated vaccine (Schankol™) allowed the mapping of continuous B-cell epitopes, topological studies, multi-antigen peptide (MAP) synthesis, and Enzyme-Linked Immunosorbent Assay (ELISA) development. Eighteen IgG epitopes were identified; eight in the CTA, three in the CTB, and seven in the protein P. Three V. cholera specific epitopes, Vc/TxA-3, Vc/TxB-11, and Vc/TxP-16, were synthesized as MAP4 and used to coat ELISA plates in order to screen immunized mouse sera. Sensitivities and specificities of 100% were obtained with the MAP4s of Vc/TxA-3 and Vc/TxB-11. The results revealed a set of peptides whose immunoreactivity reflects the immune response to vaccination. The array of peptide data can be applied to develop improved serological tests in order to detect cholera toxin exposure, as well as next generation vaccines to induce more specific antibodies against the cholera toxin.
Structure of an Alkaline Pectate Lyase and Rational Engineering with Improved Thermo-Alkaline Stability for Efficient Ramie Degumming
Authors: Zhou Cheng , Cao Yuting , Xue Yanfen , Liu Weidong , Ju Jiansong , Ma Yanhe
Published: December 2022

Alkaline pectate lyases have biotechnological applications in plant fiber processing, such as ramie degumming. Previously, we characterized an alkaline pectate lyase from Bacillus clausii S10, named BacPelA, which showed potential for enzymatic ramie degumming because of its high cleavage activity toward methylated pectins in alkaline conditions. However, BacPelA displayed poor thermo-alkaline stability. Here, we report the 1.78 Å resolution crystal structure of BacPelA in apo form. The enzyme has the characteristic right-handed β-helix fold of members of the polysaccharide lyase 1 family and shows overall structural similarity to them, but it displays some differences in the details of the secondary structure and Ca2+-binding site. On the basis of the structure, 10 sites located in flexible regions and showing high B-factor and positive ΔTm values were selected for mutation, aiming to improve the thermo-alkaline stability of the enzyme. Following site-directed saturation mutagenesis and screening, mutants A238C, R150G, and R216H showed an increase in the T5015 value at pH 10.0 of 3.0 °C, 6.5 °C, and 7.0 °C, respectively, compared with the wild-type enzyme, interestingly accompanied by a 24.5%, 46.6%, and 61.9% increase in activity. The combined mutant R150G/R216H/A238C showed an 8.5 °C increase in the T5015 value at pH 10.0, and an 86.1% increase in the specific activity at 60 °C, with approximately doubled catalytic efficiency, compared with the wild-type enzyme. Moreover, this mutant retained 86.2% activity after incubation in ramie degumming conditions (4 h, 60 °C, pH 10.0), compared with only 3.4% for wild-type BacPelA. The combined mutant increased the weight loss of ramie fibers in degumming by 30.2% compared with wild-type BacPelA. This work provides a thermo-alkaline stable, highly active pectate lyase with great potential for application in the textile industry, and also illustrates an effective strategy for rational design and improvement of pectate lyases.
Inclusion of 11-Oxygenated Androgens in a Clinical Routine LC-MS/MS Setup for Steroid Hormone Profiling
Authors: Zeidler Robert , Biemann Ronald , Ceglarek Uta , Kratzsch Jürgen , Isermann Berend , Gaudl Alexander
Published: December 2022

11-Oxygenated androgens (11-OAs) are being discussed as potential biomarkers in diagnosis and therapy control of disorders with androgen excess such as congenital adrenal hyperplasia and polycystic ovary syndrome. However, quantification of 11-OAs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) still relies on extensive sample preparation including liquid–liquid extraction, derivatization and partial long runtimes, which is unsuitable for high-throughput analysis under routine laboratory settings. For the first time, an established online-solid-phase extraction-LC-MS/MS (online-SPE-LC-MS/MS) method for the quantitation of seven serum steroids in daily routine use was extended and validated to include 11-ketoandrostenedione, 11-ketotestosterone, 11β-hydroxyandrostenedione and 11β-hydroxytestosterone. Combining a simple protein precipitation step with fast chromatographic separation and ammonium fluoride-modified ionization resulted in a high-throughput method (6.6 min run time) featuring lower limits of quantification well below endogenous ranges (63–320 pmol/L) with recoveries between 85% and 117% (CVs ≤ 15%). Furthermore, the ability of this method to distinguish between adrenal and gonadal androgens was shown by comparing 11-OAs in patients with hyperandrogenemia to healthy controls. Due to the single shot multiplex design of the method, potential clinically relevant ratios of 11-OAs and corresponding androgens were readily available. The fully validated method covering endogenous concentration levels is ready to investigate the diagnostic values of 11-OAs in prospective studies and clinical applications.
CRISPR/Cas9-Mediated Mutagenesis of BrLEAFY Delays the Bolting Time in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)
Authors: Shin Yun-Hee , Park Young-Doo
Published: December 2022

Chinese cabbage has unintended bolting in early spring due to sudden climate change. In this study, late-bolting Chinese cabbage lines were developed via mutagenesis of the BrLEAFY (BrLFY) gene, a transcription factor that determines floral identity, using the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. Double-strand break of the target region via gene editing based on nonhomologous end joining (NHEJ) was applied to acquire useful traits in plants. Based on the ‘CT001’ pseudomolecule, a single guide RNA (sgRNA) was designed and the gene-editing vector was constructed. Agrobacterium-mediated transformation was used to generate a Chinese cabbage line in which the sequence of the BrLFY paralogs was edited. In particular, single base inserted mutations occurred in the BrLFY paralogs of the LFY-7 and LFY-13 lines, and one copy of T-DNA was inserted into the intergenic region. The selected LFY-edited lines displayed continuous vegetative growth and late bolting compared to the control inbred line, ‘CT001’. Further, some LFY-edited lines showing late bolting were advanced to the next generation. The T-DNA-free E1LFY-edited lines bolted later than the inbred line, ‘CT001’. Overall, CRISPR/Cas9-mediated mutagenesis of the BrLFY gene was found to delay the bolting time. Accordingly, CRISPR/Cas9 is considered an available method for the molecular breeding of crops.
A Novel Biocidal Nanocomposite: Spherical Silica with Silver Ions Anchored at the Surface
Authors: Laskowska Magdalena , Kowalczyk Paweł , Karczmarska Agnieszka , Kramkowski Karol , Wrzosek Karol , Laskowski Łukasz
Published: December 2022

This article is devoted to a novel class of antimicrobial agents: nanocomposites composed of spherical silica and silver ions located at the silica’s surface with the assumed distribution. Such materials are in high demand due to the increasing threat from bacterial strains that are becoming resistant to currently known antibiotics. In particular, we focus on materials that make it possible to limit the growth of bacterial colonies on a variety of tactile surfaces. In this paper, we present a method for preparing a silica-based nanocomposite containing silver ions and the analysis of their antimicrobial properties. Our research revealed that the presence of tested nanocomposite induces very high oxidative stress in the bacteria cell, damaging and modifying bacterial DNA, creating oxidized guanines, cytosines, or adenines, which causes its very rapid destruction, leading to cell death.
Role of Leptin as a Link between Asthma and Obesity: A Systematic Review and Meta-Analysis
Authors: Sánchez-Ortega Helena , Jiménez-Cortegana Carlos , Novalbos-Ruiz José P. , Gómez-Bastero Ana , Soto-Campos José G. , Sánchez-Margalet Víctor
Published: December 2022

Asthma and obesity are considered as highly prevalent diseases with a great impact on public health. Obesity has been demonstrated to be an aggravating factor in the pathogenesis of asthma. Adipose tissue secretes proinflammatory cytokines and mediators, including leptin, which may promote the development and severity of asthma in obese patients. This study is a systematic review and a meta-analysis based on the relationship between leptin and asthma during obesity. MEDLINE, Cochrane, EMBASE and CINAHL databases were used. Data heterogeneity was analyzed using Cochran’s Q and treatment effect with the DerSimonian and Laird method. Random effect analyses were carried out to test data sensitivity. Asymmetry was estimated using Begg’s and Egger’s tests. All studies showed significant differences in leptin levels. The effect of the measures (p < 0.001), data sensitivity (p < 0.05) and data asymmetry were statistically significant, as well as tBegg’s test (p = 0.010) and Egge’s test (p < 0.001). Despite the existing limiting factors, the results of this study support the relevant role of leptin in the pathophysiology of asthma in obese subjects. Nevertheless, further studies are needed to obtain better insight in the relationship between leptin and asthma in obesity.
Stellate Trichomes in Dionaea muscipula Ellis (Venus Flytrap) Traps, Structure and Functions
Authors: Płachno Bartosz J. , Kapusta Małgorzata , Stolarczyk Piotr , Świątek Piotr
Published: December 2022

The digestive organs of carnivorous plants have external (abaxial) glands and trichomes, which perform various functions. Dionaea muscipula Ellis (the Venus flytrap) is a model carnivorous plant species whose traps are covered by external trichomes. The aim of the study was to fill in the gap regarding the structure of the stellate outer trichomes and their immunocytochemistry and to determine whether these data support the suggestions of other authors about the roles of these trichomes. Light and electron microscopy was used to show the trichomes’ structure. Fluorescence microscopy was used to locate the carbohydrate epitopes that are associated with the major cell wall polysaccharides and glycoproteins. The endodermal cells and internal head cells of the trichomes were differentiated as transfer cells, and this supports the idea that stellate trichomes transport solutes and are not only tomentose-like trichomes. Trichome cells differ in the composition of their cell walls, e.g., the cell walls of the internal head cells are enriched with arabinogalactan proteins (AGPs). The cell walls of the outer head cells are poor in both low and highly homogalacturonans (HGs), but the immature trichomes are rich in the pectic polysaccharide (1–4)–β-D-galactan. In the immature traps, young stellate trichomes produce mucilage which may protect the trap surface, and in particular, the trap entrance. However, the role of these trichomes is different when the outer head cells collapse. In the internal head cells, a thick secondary wall cell was deposited, which together with the thick cell walls of the outer head cells played the role of a large apoplastic space. This may suggest that mature stellate trichomes might function as hydathodes, but this should be experimentally proven.
Gelatin Meshes Enriched with Graphene Oxide and Magnetic Nanoparticles Support and Enhance the Proliferation and Neuronal Differentiation of Human Adipose-Derived Stem Cells
Authors: Șelaru Aida , Mocanu-Dobranici Alexandra-Elena , Olăreț Elena , Ginghină Raluca-Elena , Stancu Izabela-Cristina , Costache Marieta , Dinescu Sorina
Published: December 2022

The field of tissue engineering is constantly evolving due to the fabrication of novel platforms that promise to stimulate tissue regeneration in the scenario of accidents. Here, we describe the fabrication of fibrous nanostructured substrates based on fish gelatin (FG) and enriched with graphene oxide (GO) and magnetic nanoparticles (MNPs) and demonstrate its biological properties in terms of cell viability and proliferation, cell adhesion, and differentiation. For this purpose, electrospun fibers were fabricated using aqueous precursors containing either only GO and only MNP nanospecies, or both of them within a fish gelatin solution. The obtained materials were investigated in terms of morphology, aqueous media affinity, tensile elasticity, and structural characteristics. The biological evaluation was assessed against adipose-derived stem cells by MTT, LDH, Live/Dead assay, cytoskeleton investigation, and neuronal trans-differentiation. The results indicate an overall good interaction and show that these materials offer a biofriendly environment. A higher concentration of both nanospecies types induced some toxic effects, thus 0.5% GO, MNPs, and GO/MNPs turned out to be the most suitable option for biological testing. Moreover, a successful neuronal differentiation has been shown on these materials, where cells presented a typical neuronal phenotype. This study demonstrates the potential of this scaffold to be further used in tissue engineering applications.
Parahydrogen-Induced Hyperpolarization of Unsaturated Phosphoric Acid Derivatives
Authors: Zlobina Veronika V. , Kiryutin Alexey S. , Nikovskiy Igor A. , Artyushin Oleg I. , Kozinenko Vitaly P. , Peregudov Alexander S. , Yurkovskaya Alexandra V. , Novikov Valentin V.
Published: December 2022

Parahydrogen-induced nuclear polarization offers a significant increase in the sensitivity of NMR spectroscopy to create new probes for medical diagnostics by magnetic resonance imaging. As precursors of the biocompatible hyperpolarized probes, unsaturated derivatives of phosphoric acid, propargyl and allyl phosphates, are proposed. The polarization transfer to 1H and 31P nuclei of the products of their hydrogenation by parahydrogen under the ALTADENA and PASADENA conditions, and by the PH-ECHO-INEPT+ pulse sequence of NMR spectroscopy, resulted in a very high signal amplification, which is among the largest for parahydrogen-induced nuclear polarization transfer to the 31P nucleus.