bluuebunny/crossref_metadata_2025 huggingface dataset:
Teaching young adults with visual impairment and intellectual/developmental disability how to determine appropriate job fit
Authors: Kearney Kelly , Torres Ayse , Quezada Saurym
Published: July 2023

BACKGROUND: Job seekers must know how to determine whether the jobs identified during a search are a good fit. Employment decision making requires problem-solving and choice-making skills. There is limited research on employment decision making skills. OBJECTIVE: The purpose of this study was to teach three young adults with dually diagnosed visual impairment and intellectual/developmental disability to determine appropriate job fit. The mnemonic “ELSE” was paired with remote audio coaching to teach the participants how to determine if the education level, likes, skills, and earnings of each presented job were an appropriate fit for them. METHODS: This study used a single subject design to determine the effectiveness of the multicomponent intervention. The design was a multiple baseline across participant design. RESULTS: The results demonstrated all three participants were able to reach a pre-determined mastery criterion of the skill. Maintenance and generalization were variable across participants. CONCLUSIONS: All participants who received the intervention increased their ability to determine appropriate job fit. The results from this study are promising and demonstrate the potential and flexibility of the mnemonic ELSE on determining appropriate job fit by young adults with VI and IDD.
MINAT INVESTASI PEKERJA MIGRAN INDONESIA DALAM MENINGKATKAN KETAHANAN EKONOMI KELUARGA DI LAMPUNG TIMUR
Authors: Lestari Enny Puji , Putri Pertiwi Nyimas Lidya , Alimuddin Agus , Asbihani Danil
Published: July 2023

This study is to determine the investment interest of former migrant workers in improving family resilience in East Lampung and the strategies carried out by former migrant workers in increasing family economic resilience in East Lampung. Data collection by observation and interviews, data analysis techniques by collecting data on migrant worker participants in East Lampung who returned from abroad in the last 5 years at the East Lampung Manpower Office, besides that the author also collected data on former migrant workers, then reduced data and presented data. East Lampung is the 5th largest migrant worker working abroad, the investment interest of former migrant workers in improving family resilience in East Lampung is quite high, income from work is used for positive needs. In addition, factors that influence the investment interest of former migrant workers are due to interest, desire and belief. The strategy in increasing family resilience carried out by former migrant workers in East Lampung utilizes the results of work to be managed by families properly and remain productive.
ETHICAL PROBLEMS OF USING PUSH-POLLS IN MARKETING RESEARCH IN THE POLITICAL MARKET
Authors: Kushnir Taras
Published: July 2023

Thepurpose of political marketing research is to measure the electoral sentiments of voters, which are formed in the process of using various tools of influence. Critical from the ethical point of view of conducting marketing research in the political marketis their use as a tool to influence the consciousness and electoral mood of voters. Under the current conditions of marketing development, researching the needs of consumers, competitors, and macroenvironmental factors is extremely important for further planning of the company's activities. Marketing research, in addition to the fact that it should help in making effective management decisions, can be used by unscrupulous customers for various manipulations. For example, in the conditions of the industrialgoods market, such manipulations are practically unreal, because each of the participants systematically makes purchasing decisions and is very well acquainted with both the market participants and the technical and functional characteristics of the goodsthat are the subject of procurement. In the political market, the "purchase of goods" is carried out regularly, but with large time intervals (once every 4 or 5 years), which, given the unformed political culture, creates extremely favorable conditions for the use of various manipulations and technologies to influence the voter (consumer of political goods).The main methods of the conducted research are the analysis, generalization and processing of information obtained as a result of sociological research conducted by the Batkivshchyna party, the sociological company Rating, the Kyiv International Institute of Sociology, and the Razumkov Center. The method of correlation analysis was also used to identify the relationship between the poll results and the actual voting results. As a result of the analysis, it was found that the use of manipulative technologies in the process of conducting marketing political research in the conditions of the pre-election process is ineffective and difficult to predict from the point of view of the final result. Thus, it can be argued that the Ukrainian voter is influenced by a complex of marketing and technological means and techniques, and individual efforts do not bring the desired result
GLOBAL CITIES AS CENTRES OF GLOBAL ECONOMIC MANAGEMENT AND INTERNATIONAL INFORMATION COMMUNICATIONS
Authors: Shklyar Viktoriya , Strilok Ivanna , Kuzmenko Oksana
Published: July 2023

Global cities arebecoming increasingly important players in the global system. They bring together different cultures, ideas and business opportunities. Understanding their role and functions in global economic governance and international information exchange is key to developing strategies for economic growth, creating favourable conditions for doing business and increasing the competitiveness of countries.The study of this problem has been based on the consideration of various aspects, such as economic activity, transnational governance, information communications, technological development and political participation, which determine the role of global cities. Using methods of comparative analysis, sociological, geographical research and other approaches, the article offers a deeper understanding of the functions of global cities and their impact on the world economy and international information communications.The functional significance of global cities as centres of global economic governance is determined mainly by the scale of their gross regional product, huge economic potential, as well as management and control of the global economy, which is due to the level of concentration of headquarters of large multinational companies, key institutions of the international financial market, mainly transnational banks, advanced business services companies, etc.Global cities are the backbone of the global economic and political system, leading global financial and economic processes, managing global financial flows, and having developed infrastructure and access to a wide range of services. Multinational enterprises actively use the advantages of the transport, political, social and economic position of global cities. In turn, these enterprises have a significant impact on global economic and political processes, contributing to the globalisation of city functions.Since the functional significance of global cities is expressed in many aspects, based on the analysis of available data, reports and studies, they are systematised into two groups: centres of global economic management and centres of international information communications. The characteristic of global cities according to these groups is due to the constant development and expansion of strong ties between countries and the destruction of existing barriers through technological communications and networking.
THREATS TO THE TAX SECURITY OF TAXPAYERS IN THE MODERN ECONOMIC CONDITIONS
Authors: Shulyarenko Svіtlana
Published: July 2023

Taxes are the main source of filling the state budget. Considerable attention is paid to the fiscal security of the state in the conditions of martial law. Ensuring the country's tax security is impossible without compliance with tax security by the taxpayers themselves. Scientists consider tax security at different levels: tax security of taxpayers, the region, the state, international tax security. Tax security consists of such elements as threats, risks, level, indicators of tax security. The article substantiates the concept of tax security of taxpayers and tax security management as a set of measures to ensure the optimal level of tax security. The tasks of tax security management include identifying threats to tax security, determining the actual and optimal level of tax security, substantiating and implementing measures to ensure the optimal level of tax security, determining the effectiveness of measures to adjust management decisions. Threats to the tax security of taxpayers are divided into managed and unmanaged by the possibility of influencing and managing them. The managed types of threats include the company's management system, the qualifications of accounting staff, the level of tax culture, accounting policy, and the taxpayer's financial condition; to the unmanaged -the system of administration of taxes and fees, changes in tax legislation and the structure of control bodies, a high level of tax burden, force majeure, circumstances of force majeure. Blocking the registration of tax invoices in the Unified register of tax invoices as a tool for electronic administration of value added tax poses a threat to VAT payers, leads to the impossibility of recognizing the tax liability and VAT tax credit. The introduced changes in taxation had a positive effect on taxpayers and the country's budget during martial law.Keywords:tax security, tax security management, threats to tax security, administration of taxes and fees.
MODERN CHALLENGES OF THE THEORY AND PRACTICE OF INTERNATIONAL COMMUNICATIONS IN THE SYSTEMOF ADMINISTRATIVE MANAGEMENT OF THE ORGANIZATION IN THE CONDITIONS OF SUSTAINABLE DEVELOPMENT: A SYSTEMATIC APPROACH
Authors: Batsenko Liudmyla , Halenin Roman
Published: July 2023

The article examines the current problem of international communications of administrative management in the conditions of sustainabledevelopment, and also substantiates the need to apply a systemic approach to its solution. The world is becoming increasingly globalized, leading to a growing need for improving communication processes between organizations from different countries and cultures. To achieve the articles’ goal, the main approaches to system analysis were considered, as well as their application to improve international communications. The role of the system approach in increasing the efficiency of interaction between variousstakeholders and promoting sustainable development is analyzed in detail.
Additionally, the article provides specific recommendations for the application of systemic analysis methods in practical situations of international communications. This may include implementing a monitoring and evaluation system for communication strategies, developing culturally adaptive approaches to engage with various stakeholders, and utilizing intercultural training and education tools to enhance perception and understandingof different cultural contexts.The concluding remarks of the article emphasize that the use of a systemic approach is essential for effectively addressing the challenges of international communications in the context of sustainable development. It facilitates collaboration among stakeholders, improves the qualityof communication processes, and contributes to achieving sustainable development on an international level.The article contains valuable information for professionals in the field of administrative management who are interested in enhancing international communications for the purpose of sustainable development. It provides a comprehensive understanding of the application of a systemic approach in this context and offers specific methods and recommendations for their practical implementation.
Duración razonable del proceso tributario
Authors: Blatt Daniel Soriano
Published: July 2023

Este artículo tiene como objetivo ver el papel del principio de duración razonable del proceso en el ámbito constitucional y procesal tributario, así como su interconexión con el campo de los Derechos Humanos. El principio de la duración razonable del proceso se fundamenta en varios diplomas jurídicos, entre ellos la Declaración Universal de los Derechos Humanos y la Convención Americana sobre Derechos Humanos. Se realizó una comparación con otros ordenamientos jurídicos, como el italiano y el español, donde se pudo comprobar que dicho principio forma parte de sus respectivos textos constitucionales. En Brasil, el principio de la duración razonable de los procesos está respaldado por la Constitución Federal, así como por la legislación infraconstitucional. Finalmente, se pudo comprobar que, a pesar de los avances, las normas existentes combaten la lentitud de las autoridades fiscales, pero no la inercia del Poder Judicial, lo que lleva a constantes violaciones a este principio fundamental en el ámbito tributario y en el mundo del Derecho como entero.
E-SAMSAT: SARANA UNTUK MEMBANTU DAN MEMUDAHKAN WAJIB PAJAK MELAKUKAN PEMBAYARAN PAJAK KENDARAAN BERMOTOR
Authors: Rindiyani Rina , Faisol Moh.
Published: July 2023

The aims of this reseach to know the implementation of E-Samsat at the Sumenep District SAMSAT Office. The researcher uses the case study method by conducting interviews with five informants: one SAMSAT office employee and four motor vehicle taxpayers who use E-Samsat. To analyze the data using qualitative data analysis (Miles & Huberman). The results of this study state that the implementation of the use of E-Samsat is very helpful and makes it easier for taxpayers to make motor vehicle tax payments. Taxpayers don't need to stand in long queues, because they can make tax payments from anywhere, and at any time via E-Samsat. However, the biggest contribution to accessing E-Samsat is the internet network, so whether or not the network is stable affects the utilization of E-Samsat.
Image recognition of traditional Chinese medicine based on deep learning
Authors: Miao Junfeng , Huang Yanan , Wang Zhaoshun , Wu Zeqing , Lv Jianhui
Published: July 2023

Chinese herbal medicine is an essential part of traditional Chinese medicine and herbalism, and has important significance in the treatment combined with modern medicine. The correct use of Chinese herbal medicine, including identification and classification, is crucial to the life safety of patients. Recently, deep learning has achieved advanced performance in image classification, and researchers have applied this technology to carry out classification work on traditional Chinese medicine and its products. Therefore, this paper uses the improved ConvNeXt network to extract features and classify traditional Chinese medicine. Its structure is to fuse ConvNeXt with ACMix network to improve the performance of ConvNeXt feature extraction. Through using data processing and data augmentation techniques, the sample size is indirectly expanded, the generalization ability is enhanced, and the feature extraction ability is improved. A traditional Chinese medicine classification model is established, and the good recognition results are achieved. Finally, the effectiveness of traditional Chinese medicine identification is verified through the established classification model, and different depth of network models are compared to improve the efficiency and accuracy of the model.
Molecular characterization of multidrug resistant Acinetobacter baumannii clinical isolates from Alexandria, Egypt
Authors: Sánchez-Urtaza Sandra , Ocampo-Sosa Alain , Molins-Bengoetxea Ainhoa , El-Kholy Mohammed A. , Hernandez Marta , Abad David , Shawky Sherine M. , Alkorta Itziar , Gallego Lucia
Published: July 2023

Carbapenem resistant Acinetobacter baumannii is a major global concern, especially in countries of the Middle East and North Africa, where the antibiotic resistance rates are on the rise. The aim of this study was to study the genomic characteristics and antimicrobial susceptibility profile of thirty-six multidrug resistant A. baumannii clinical isolates obtained in hospitals from Alexandria, Egypt. Antibiotic resistance rates were estimated by determination of Minimum Inhibitory Concentrations. Carbapenemase genes, other antibiotic resistance genes and virulence factors were then screened by the use of Whole Genome Sequencing. Isolates were also subjected to Multi Locus Sequence Typing (MLST) using the Pasteur Scheme and to core genome MLST to study their clonal relatedness. In addition, plasmid analysis was performed by the use of a commercial kit and S1- Pulsed Field Gel Electrophoresis, and Hybridization experiments with DIG-labeled DNA probes for blaNDM-1, blaPER-7 and blaGES-like were performed to locate these genes. The majority of isolates were resistant to β-lactams (including carbapenems), fluoroquinolones, aminoglycosides and trimethoprim; and some showed resistance to cefiderocol and minocycline. We identified 8 different blaOXA-51-like variants including blaOXA-51, blaOXA-64, blaOXA-65, blaOXA-66, blaOXA-68, blaOXA-91, blaOXA-94 and blaOXA-336; blaOXA-23, blaNDM-1, blaPER-7, blaGES-like and blaADC-like and other antibiotic resistance genes, some of these genes were within transposons or class 1 integrons. Multiple virulence factors responsible for adherence, biofilm production, type II and type VI secretion systems, exotoxins, exoenzymes, immune modulation and iron uptake were observed and 34 out of 36 isolates showed motility. Thirty-five out of 36 isolates clustered with International Clones 2, 4, 5, 7, 8 and 9; and 9 STs were identified including ST570, ST2, ST600, ST15, ST113, ST613, ST85, ST158, ST164. Plasmids ranging in size from 1.7 to 70 kb were found; blaNDM-1 and blaPER-7 genes were located in the chromosome and blaGES-like genes were simultaneously located in the chromosome and in a plasmid of 70kb. In conclusion, this study revealed a wide spectrum of antibiotic resistance genes and a variety of lineages among A. baumannii isolated in hospitals from Alexandria, and highlights the importance of investigating the molecular epidemiology to control the spread of multi-drug resistant isolates.
When amiodarone-induced thyroiditis meets cardiomyopathy with excessive trabeculation: a case report
Authors: Gašparini Dora , Raljević Damir , Pehar-Pejčinović Vesna , Klarica Gembić Tihana , Peršić Viktor , Turk Wensveen Tamara
Published: July 2023

Introduction

Amiodarone is a potent antiarrhythmic medication used to treat life-threatening ventricular arrhythmias; however, its well-established adverse effect is a thyroid disorder. Amiodarone-induced thyroiditis (AIT), a clinical entity involving two types with different etiopathology and treatment approaches, may occur at the beginning or even several years after amiodarone treatment discontinuation. The toxicity profile of amiodarone becomes especially important in young patients with lifelong cardiac disorders, which are often refractory to other antiarrhythmic drugs. Herein, we report the first case of non-sustained ventricular tachycardia (NSVT), an unusual presentation of type II AIT, in a young male patient who was previously diagnosed with left ventricular cardiomyopathy with excessive trabeculation.

Case report

A 36-year-old male non-athlete presented with tiredness during regular follow-up. Continuous electrocardiographic monitoring (cECG) revealed NSVT, whereas echocardiography and cardiac magnetic resonance imaging detected discrete structural and functional changes that could not fully explain the observed cECG report. Conversely, an unmeasurably low thyrotropin level on admission and previous exposure to amiodarone pointed the diagnostic pathway in the direction of the thyroid gland. Elevated free thyroxine and undetectable autoantibody titers with unremarkable sonographic findings raised clinical suspicion of type II AIT. Scintigraphic imaging with 99mTc-2-methoxyisobutylisonitrile (sestamibi) revealed decreased thyroid uptake; hence, prednisone was introduced for treatment. Clear improvements in both biochemical and electrocardiographic parameters were observed after immunomodulatory treatment of type II AIT in this young patient with cardiomyopathy and excessive trabeculation.

Conclusion

Treatment of reversible causes of cardiac rhythm abnormalities such as type II AIT should be considered before choosing other treatment modalities, particularly in patients with structural cardiac disorders. The importance of a multidisciplinary approach in complex cases such as the one reported, thus, cannot be emphasized enough.
Too much information is no information: how machine learning and feature selection could help in understanding the motor control of pointing
Authors: Thomas Elizabeth , Ali Ferid Ben , Tolambiya Arvind , Chambellant Florian , Gaveau Jérémie
Published: July 2023

The aim of this study was to develop the use of Machine Learning techniques as a means of multivariate analysis in studies of motor control. These studies generate a huge amount of data, the analysis of which continues to be largely univariate. We propose the use of machine learning classification and feature selection as a means of uncovering feature combinations that are altered between conditions. High dimensional electromyogram (EMG) vectors were generated as several arm and trunk muscles were recorded while subjects pointed at various angles above and below the gravity neutral horizontal plane. We used Linear Discriminant Analysis (LDA) to carry out binary classifications between the EMG vectors for pointing at a particular angle, vs. pointing at the gravity neutral direction. Classification success provided a composite index of muscular adjustments for various task constraints—in this case, pointing angles. In order to find the combination of features that were significantly altered between task conditions, we conducted a post classification feature selection i.e., investigated which combination of features had allowed for the classification. Feature selection was done by comparing the representations of each category created by LDA for the classification. In other words computing the difference between the representations of each class. We propose that this approach will help with comparing high dimensional EMG patterns in two ways; (i) quantifying the effects of the entire pattern rather than using single arbitrarily defined variables and (ii) identifying the parts of the patterns that convey the most information regarding the investigated effects.
Down-staging of obesity one year after bariatric surgery: a new proposal of Edmonton obesity staging system
Authors: Quinto Giulia , Bettini Silvia , Neunhaeuserer Daniel , Battista Francesca , Milan Gabriella , Gasperetti Andrea , Vecchiato Marco , Vettor Roberto , Ermolao Andrea , Busetto Luca
Published: July 2023

Background

Different approaches are used to classify obesity severity. The Edmonton Obesity Staging System (EOSS) considers medical, physical and psychological parameters. A new modified EOSS with a different functional evaluation method, measuring Cardiorespiratory Fitness (CRF), has been recently proposed, EOSS-CRF. Bariatric surgery (BS) is one of the most efficient treatments of obesity and all aspect of related disorders. No studies have yet applied EOSS-CRF after BS. Therefore, the aim of this study was to evaluate modifications in EOSS and EOSS-CRF before and after BS.

Methods

This observational study finally enrolled 72 patients affected by obesity. A multi-disciplinary assessment in order to evaluate eligibility to surgical treatment has been performed, including anamnesis, physical evaluation, anthropometric data measurement, biochemical blood exams and cardiopulmonary exercise testing. One year after BS the same protocol was applied. Patients have been classified according to EOSS and EOSS-CRF before and one year after BS.

Results

After BS, patients categorized in classes associated to severe obesity (EOSS ≥ 2 or EOSS-CRF ≥ 2) reduced significantly. Using EOSS, patients without functional impairment were 61% before surgery and 69% after BS (p=0.383). Using EOSS-CRF, patients considered without functional impairment were only 9.7% before BS; this percentage significantly raised to 50% after BS (p<0.001). The impact of functional domains before and after BS is different in grading patients in EOSS and EOSS-CRF, respectively.

Conclusions

Improvements obtained after BS are adequately summarized by EOSS and EOSS-CRF. The EOSS-CRF grading method for functional impairment seems to better reflect the known amelioration obtained after BS. Objective measurements of CRF may provide additional value to classify severity of obesity, also in the follow-up after BS.
Prostaglandin F2α requires activation of calcium-dependent signalling to trigger inflammation in human myometrium
Authors: Riaposova Lucia , Kim Sung Hye , Hanyaloglu Aylin C. , Sykes Lynne , MacIntyre David A. , Bennett Phillip R. , Terzidou Vasso
Published: July 2023

Introduction

Preterm birth is one of the major causes of neonatal morbidity and mortality across the world. Both term and preterm labour are preceded by inflammatory activation in uterine tissues. This includes increased leukocyte infiltration, and subsequent increase in chemokine and cytokine levels, activation of pro-inflammatory transcription factors as NF-κB and increased prostaglandin synthesis. Prostaglandin F2α (PGF2α) is one of the myometrial activators and stimulators.

Methods

Here we investigated the role of PGF2α in pro-inflammatory signalling pathways in human myometrial cells isolated from term non-labouring uterine tissue. Primary myometrial cells were treated with G protein inhibitors, calcium chelators and/or PGF2α. Nuclear extracts were analysed by TranSignal cAMP/Calcium Protein/DNA Array. Whole cell protein lysates were analysed by Western blotting. mRNA levels of target genes were analysed by RT-PCR.

Results

The results show that PGF2α increases inflammation in myometrial cells through increased activation of NF-κB and MAP kinases and increased expression of COX-2. PGF2α was found to activate several calcium/cAMP-dependent transcription factors, such as CREB and C/EBP-β. mRNA levels of NF-κB-regulated cytokines and chemokines were also elevated with PGF2α stimulation. We have shown that the increase in PGF2α-mediated COX-2 expression in myometrial cells requires coupling of the FP receptor to both Gαq and Gαi proteins. Additionally, PGF2α-induced calcium response was also mediated through Gαq and Gαi coupling.

Discussion

In summary, our findings suggest that PGF2α-induced inflammation in myometrial cells involves activation of several transcription factors – NF-κB, MAP kinases, CREB and C/EBP-β. Our results indicate that the FP receptor signals via Gαq and Gαi coupling in myometrium. This work provides insight into PGF2α pro-inflammatory signalling in term myometrium prior to the onset of labour and suggests that PGF2α signalling pathways could be a potential target for management of preterm labour.
Five-year measurement data along a 1200 m elevational gradient reveals that global warming increases soil respiration
Authors: Takeda Soichiro , Majima Ryota , Makita Naoki , Takahashi Koichi
Published: July 2023

Soil respiration is a major pathway for CO2 emissions from ecosystems. Owing to its temperature dependency, the soil respiration rate is expected to increase due to global warming, particularly at high elevations. To clarify the effects of soil temperature and volumetric soil water content on soil respiration rates (RS), we examined seasonal changes in RS at five elevations of 1600–2800 m in subalpine coniferous forests in Japan for 5 years. The aboveground biomass of forest stands decreased from 282 to 29 Mg/ha as elevation increased. The monthly mean RS was lower at higher elevations from July to October. While RS was positively correlated with soil temperature at each elevation, the effect of soil water content on RS varied among the five elevations. Seasonal changes in RS could be reproduced from soil temperature and soil water content for each elevation in each year. RS at any temperature was lower at higher elevations because RS was also positively correlated with aboveground biomass. From 1600 to 2800 m, the annual RS was estimated to decrease from 2.79 to 0.74 kg CO2 year–1 m–2. The annual RS along the elevational gradient was predicted to increase by 9–12% and 30–42% under low and high greenhouse gas emission scenarios (annual mean temperature 0.76°C and 3.3°C increases), respectively, during 2095–2100 compared to the current period 2015–2020. Increased soil respiration rate will accelerate global warming via the positive feedback. Overall, our findings suggest that soil respiration evaluation is important not only for calculating the carbon balance of forest stands due to global warming but also for predicting global warming owing to the feedback of CO2 emission from soil to atmosphere.
Establishment of a genome editing tool using CRISPR-Cas9 ribonucleoprotein complexes in the non-model plant pathogen Sphaerulina musiva
Authors: Tannous Joanna , Sawyer Cole , Hassan Md Mahmudul , Labbe Jesse L. , Eckert Carrie
Published: July 2023

CRISPR-Cas9 is a versatile genome editing system widely used since 2013 to introduce site-specific modifications into the genomes of model and non-model species. This technology is used in various applications, from gene knock-outs, knock-ins, and over-expressions to more precise changes, such as the introduction of nucleotides at a targeted locus. CRISPR-Cas9 has been demonstrated to be easy to establish in new species and highly efficient and specific compared to previous gene editing strategies such as Zinc finger nucleases and transcription activator-like effector nucleases. Grand challenges for emerging CRISPR-Cas9 tools in filamentous fungi are developing efficient transformation methods for non-model organisms. In this paper, we have leveraged the establishment of CRISPR-Cas9 genome editing tool that relies on Cas9/sgRNA ribonucleoprotein complexes (RNPs) in the model species Trichoderma reesei and developed the first protocol to efficiently transform the non-model species, Sphaerulina musiva. This fungal pathogen constitutes a real threat to the genus Populus, a foundational bioenergy crop used for biofuel production. Herein, we highlight the general considerations to design sgRNAs and their computational validation. We also describe the use of isolated protoplasts to deliver the CRISPR-Cas9 RNP components in both species and the screening for targeted genome editing events. The development of engineering tools in S. musiva can be used for studying genes involved in diverse processes such as secondary metabolism, establishment, and pathogenicity, among many others, but also for developing genetic mitigation approaches. The approach described here provides guidance for potential development of transformation systems in other non-model spore-bearing ascomycetes.
Genetic manipulation of betta fish
Authors: Palmiotti Alec , Lichak Madison R. , Shih Pei-Yin , Kwon Young Mi , Bendesky Andres
Published: July 2023

Betta splendens, also known as Siamese fighting fish or “betta,” is a freshwater fish species renowned for its astonishing morphological diversity and extreme aggressive behavior. Despite recent advances in our understanding of the genetics and neurobiology of betta, the lack of tools to manipulate their genome has hindered progress at functional and mechanistic levels. In this study, we outline the use of three genetic manipulation technologies, which we have optimized for use in betta: CRISPR/Cas9-mediated knockout, CRISPR/Cas9-mediated knockin, and Tol2-mediated transgenesis. We knocked out three genes: alkal2l, bco1l, and mitfa, and analyzed their effects on viability and pigmentation. Furthermore, we knocked in a fluorescent protein into the mitfa locus, a proof-of-principle experiment of this powerful technology in betta. Finally, we used Tol2-mediated transgenesis to create fish with ubiquitous expression of GFP, and then developed a bicistronic plasmid with heart-specific expression of a red fluorescent protein to serve as a visible marker of successful transgenesis. Our work highlights the potential for the genetic manipulation of betta, providing valuable resources for the effective use of genetic tools in this animal model.
A novel 3D spheroid model of rheumatoid arthritis synovial tissue incorporating fibroblasts, endothelial cells, and macrophages
Authors: Philippon Eva M. L. , van Rooijen Lisanne J. E. , Khodadust Fatemeh , van Hamburg Jan Piet , van der Laken Conny J. , Tas Sander W.
Published: July 2023

Objective

Rheumatoid Arthritis (RA) is a progressive and systemic autoimmune disorder associated with chronic and destructive joint inflammation. The hallmarks of joint synovial inflammation are cellular proliferation, extensive neoangiogenesis and infiltration of immune cells, including macrophages. In vitro approaches simulating RA synovial tissue are crucial in preclinical and translational research to evaluate novel diagnostic and/or therapeutic markers. Two-dimensional (2D) settings present very limited in vivo physiological proximity as they cannot recapitulate cell-cell and cell-matrix interactions occurring in the three-dimensional (3D) tissue compartment. Here, we present the engineering of a spheroid-based model of RA synovial tissue which mimics 3D interactions between cells and pro-inflammatory mediators present in the inflamed synovium.

Methods

Spheroids were generated by culturing RA fibroblast-like-synoviocytes (RAFLS), human umbilical vein endothelial cells (ECs) and monocyte-derived macrophages in a collagen-based 3D scaffold. The spheroids were cultured in the presence or absence of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (bFGF) or RA synovial fluid (SF). Spheroid expansion and cell migration were quantified for all conditions using confocal microscopy and digital image analysis.

Results

A novel approach using machine learning was developed to quantify spheroid outgrowth and used to reexamine the existing spheroid-based model of RA synovial angiogenesis consisting of ECs and RAFLS. A 2-fold increase in the spheroid outgrowth ratio was demonstrated upon VEGF/bFGF stimulation (p<0.05). The addition of macrophages within the spheroid structure (3.75x104 RAFLS, 7.5x104 ECs and 3.0x104 macrophages) resulted in good incorporation of the new cell type. The addition of VEGF/bFGF significantly induced spheroid outgrowth (p<0.05) in the new system. SF stimulation enhanced containment of macrophages within the spheroids.

Conclusion

We present a novel spheroid based model consisting of RAFLS, ECs and macrophages that reflects the RA synovial tissue microenvironment. This model may be used to dissect the role of specific cell types in inflammatory responses in RA, to study specific signaling pathways involved in the disease pathogenesis and examine the effects of novel diagnostic (molecular imaging) and therapeutic compounds, including small molecule inhibitors and biologics.
Suppressive myeloid cells in SARS-CoV-2 and Mycobacterium tuberculosis co-infection
Authors: Shaw Jane Alexandra , Malherbe Stephanus T. , Walzl Gerhard , du Plessis Nelita
Published: July 2023

Epidemiologic data show that both current and previous tuberculosis (TB) increase the risk of in-hospital mortality from coronavirus disease-2019 (COVID-19), and there is a similar trend for poor outcomes from Mycobacterium tuberculosis (Mtb) infection after recent SARS-CoV-2. A shared dysregulation of immunity explains the dual risk posed by co-infection, but the specific mechanisms are being explored. While initial attention focused on T cell immunity, more comprehensive analyses revealed a dysfunctional innate immune response in COVID-19, characterized by reduced numbers of dendritic cells, NK cells and a redistribution of mononuclear phagocytes towards intermediate myeloid subsets. During hyper- or chronic inflammatory processes, activation signals from molecules such as growth factors and alarmins lead to the expansion of an immature population of myeloid cells called myeloid-deprived suppressor cells (MDSC). These cells enter a state of pathological activation, lose their ability to rapidly clear pathogens, and instead become broadly immunosuppressive. MDSC are enriched in the peripheral blood of patients with severe COVID-19; associated with mortality; and with higher levels of inflammatory cytokines. In TB, MDSC have been implicated in loss of control of Mtb in the granuloma and ineffective innate and T cell immunity to the pathogen. Considering that innate immune sensing serves as first line of both anti-bacterial and anti-viral defence mechanisms, we propose MDSC as a crucial mechanism for the adverse clinical trajectories of TB-COVID-19 coinfection.
Trophic structure and fishing impacts on an oligotrophic ecosystem in the Western Mediterranean: the Balearic Islands
Authors: Sánchez-Zulueta Paula , Valls María , Guijarro Beatriz , Ángeles Torres María , Ángeles Zapata María , Coll Marta , Corrales Xavier , Andonegi Eider , Díaz-Valdés Marta , Massutí Enric , Ordines Francesc
Published: July 2023

Within the context of the ecosystem approach to fisheries management, an ecosystem model was developed for the Balearic Islands in the early 2000s, covering from 0 to 800 m of depth. The aim of the study was to describe the structure and functioning of the ecosystem together with the fishing impacts. The results show that the biomass of the primary producers (PP), mainly dominated by Posidonia oceanica meadows and red algae beds, represented a high percentage of the biomass (39.20%) in the ecosystem. Most of the trophic flows occurred between PP (mostly benthic) and trophic level II. The mixed trophic impact analysis also highlighted a positive impact of the functional groups (FGs) at the base of the food web. Besides, there were important trophic flows between several demersal FGs, and many groups of the pelagic compartment, confirming the importance of the benthopelagic coupling previously reported. The mean trophic level of the catch (mTLc=2.76) was lower than those reported in adjacent areas due to the discard of benthic producers. By excluding benthic PP, the Balearic Islands showed the highest mTLc (3.44) among all the Mediterranean areas modelled, probably due to differences in the trophic structure, lower fisheries exploitation levels and lower development of the purse seine fleet targeting small pelagic fishes. The mean transfer efficiency (21.80%) was double than the global average (10%), which highlights the oligotrophy of the system, the importance of demersal production which has lower rates of transfer through the food web, and a more efficient use of nutrients in the ecosystem. Sponges, one of the most important groups of benthic invertebrates in the Balearic Archipelago, are known to recycle dissolved organic matter, which is reintroduced into the food web similar to the microbial loop, but the magnitude of this and other processes remains still unknown in this area. The present study is a first step to develop spatio-temporal simulations under different exploitation scenarios and to calculate ecological indicators to assess the state of the marine environment in this region.
Fabrication of bioactive nanocomposites from chitosan, cress mucilage, and selenium nanoparticles with powerful antibacterial and anticancerous actions
Authors: El-Sherbiny Mohsen M. , Orif Mohamed I. , El-Hefnawy Mohamed E. , Alhayyani Sultan , Al-Goul Soha T. , Elekhtiar Rawan S. , Mahrous Hoda , Tayel Ahmed A.
Published: July 2023

Natural bioactive alternatives are the utmost requests from researchers to provide biosafe and effectual health-guarding agents. The biopolymers chitosan nanoparticles (NCT), mucilage of cress seed (GCm; Lepidium sativum), and GCm-mediated selenium nanoparticles (GCm/SeNPs) were innovatively employed for fabricating novel bioactive natural nanocomposites (NCs) with elevated bioactivities as bactericidal (against Salmonella typhimurium and Staphylococcus aureus) and anticancer (against CaCo-2 and HeLa cells). The SeNPs were successfully generated with GCm, and different NCs formulations were fabricated from NCT:GCm/SeNPs amalgam ratios including T1, T2, and T3 with 2:1, 1:1, and 1:2 ratios, respectively. The infrared analysis of synthesized molecules appointed apparent physical interactions among interacted molecules. The average particles’ sizes and charges of molecules/NCs were (12.7, 316.4, 252.8, and 127.3 nm) and (−6.9, +38.7, +26.2, and −25.8 mV) for SeNPs, T1, T2, and T3, respectively. The biocidal assessment of NCs indicated that T1 was the strongest antibacterial formulation, whereas T3 was the superior anticancer amalgam. These NCs formulations could exceed the biocidal potentialities of standard biocides. T1-NC could cause severe destructions/deformations in challenged S. typhimurium within 9 h, whereas T3-NCs induced apparent fluorescent apoptosis signs in treated HeLa cells. The prospective applications innovatively designed biocidal natural NCs that are recommended for controlling pathogenic bacteria and fighting cancerous cells.
Potential biocide roles of violacein
Authors: Rivero Berti Ignacio , Gantner Melisa E. , Rodriguez Santiago , Islan German A. , Fávaro Wagner J. , Talevi Alan , Castro Guillermo R. , Durán Nelson
Published: July 2023

Violacein is a pigment produced by Gram-negative bacteria, which has shown several beneficial biological activities. The most relevant activities of violacein include the interference in the physiological activities of biological membranes, inhibition of cell proliferation, antioxidant, and anti-inflammatory activities. Moreover, the antiviral activities of violacein against some enveloped and non-enveloped viruses have also been reported. Violacein showed a wide spectrum of protease inhibition, both experimentally and in silico. Other in silico studies have suggested that violacein binds to the SARS-CoV-2 spike. Empirical physicochemical studies indicate that violacein (or, occasionally, its derivatives) may be administered orally to treat different disorders. In addition, different alternatives to product violacein, and molecular devices for delivery of this pigment are reviewed.
Unlocking the enigma: unraveling multiple cognitive dysfunction linked to glymphatic impairment in early Alzheimer’s disease
Authors: Zhong Jiayi , Zhang Xiaochen , Xu Huanyu , Zheng Xiaoran , Wang Luyao , Jiang Jiehui , Li Yunxia
Published: July 2023

Background

Alzheimer’s disease (AD) is one of the world’s well-known neurodegenerative diseases, which is related to the balance mechanism of production and clearance of two proteins (amyloid-β and tau) regulated by the glymphatic system. Latest studies have found that AD patients exhibit impairments to their glymphatic system. However, the alterations in the AD disease continuum, especially in the early stages, remain unclear. Moreover, the relationship between the glymphatic system and cognitive dysfunction is still worth exploring.

Methods

A novel diffusion tensor image analysis method was applied to evaluate the activity of the glymphatic system by an index for diffusivity along the perivascular space (ALPS-index). Based on this method, the activity of the glymphatic system was noninvasively evaluated in 300 subjects, including 111 normal controls (NC), 120 subjects with mild cognitive impairment (MCI), and 69 subjects with AD. Partial correlation analysis was applied to explore the association between glymphatic system and cognitive impairment based on three domain-general scales and several domain-specific cognitive scales. Receiver operating characteristic curve analysis was used to evaluate the classification performance of ALPS-index along the AD continuum.

Results

ALPS-index was significantly different among NC, MCI and AD groups, and ALPS-index decreased with cognitive decline. In addition, ALPS-index was significantly correlated with the scores of the clinical scales (p<0.05, FDR corrected), especially in left hemisphere. Furthermore, combination of ALPS and fractional anisotropy (FA) values achieved better classification results (NC vs. MCI: AUC = 0.6610, NC vs. AD: AUC = 0.8214).

Conclusion

Here, we show that the glymphatic system is closely associated with multiple cognitive dysfunctions, and ALPS-index can be used as a biomarker for alterations along the AD continuum. This may provide new targets and strategies for the treatment of AD, and has the potential to assist clinical diagnosis.
Amino acid intake with protein food source and incident dyslipidemia in Korean adults from the Ansan and Ansung Study and the Health Examinee Study
Authors: Chung Sangwon , Park Jae Ho , Joung Hyojee , Ha Kyungho , Shin Sangah
Published: July 2023

Background

Dyslipidemia is a major risk factor for cardiovascular diseases and appropriate intake of amino acids may be helpful for the management of dyslipidemia. However, evidence of an association between amino acid intake and dyslipidemia in Korean adults is limited.

Objective

The purpose of this study was to investigate how the incidence of dyslipidemia in Korean adults is associated with the consumption of amino acids, essential and nonessential types, as well as the sources of these amino acids from food.

Methods

Data from 35,478 study participants without dyslipidemia at baseline from the Ansan and Ansung Study and the Health Examinee Study were used for the analysis. Dyslipidemia and its components such as hypertriglyceridemia, hypercholesterolemia, hyper-low-density lipoprotein (LDL) cholesterolemia and hypo-high-density lipoprotein (HDL) cholesterolemia were the main outcome in this study. The participants were categorized into quartiles, based on the intake of amino acids and plant−/animal-based proteins.

Results

On average, the follow-up period lasted for 5.7 years. The two major food groups that contributed to one-half of the intake for each type of amino acid were whole grain mixed rice and white rice. Compared to the lowest quartile group, the highest quartile groups of essential amino acid intake [men: hazard ratio (HR) = 0.78; 95% confidence interval (CI), 0.63–0.97; P for trend = 0.0088; women: HR = 0.86; 95% CI, 0.76–0.99; P for trend = 0.0201] and nonessential amino acid intake (men: HR = 0.75; 95% CI, 0.60–0.94; P for trend = 0.0069; women: HR = 0.81; 95% CI, 0.71–0.93; P for trend = 0.0024) had a decreased risk of dyslipidemia. Plant-based protein intake had a negative association and animal-based protein intake had a nonsignificant association with dyslipidemia after adjustment for energy-adjusted fat intake. Furthermore, the essential and nonessential amino acid intake showed stronger negative associations with dyslipidemia after further adjustment for energy-adjusted fat intake.

Conclusion

To conclude, the intake of amino acids may have a protective effect against dyslipidemia in Korean adults who are aged 40 years or older, regardless of their protein food sources.