bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and Expression Pattern of the Carotenoid Cleavage Oxygenase Gene Family in Lycium Suggest CCOs Respond to Abiotic Stress and Promote Carotenoids Degradation
Authors: Li Weinan , Che Jiahang , Lian Qile , Wang Cuiping , Dai Guoli , Chen Jinhuan
Published: May 2023

Carotenoids are key metabolites in goji (Lycium), a traditional Chinese medicine plant; however, the carotenoid content varies in fruits of different goji species, and the mechanism of this variation is not clear. Carotenoids participate in signal transduction and photosynthesis, and function as colorants and photoprotectors. Members of the carotenoid cleavage oxygenase (CCO) gene family are involved in the regulation of phytohormones, pigments, and aromatic substances, such as abscisic acid (ABA), β-carotenoid, and α-ionone, by degrading carotenoids. Some CCO genes are also related to an abiotic stress response. Here, a total of 12 LbCCO genes were identified and analyzed from the L. barbarum genome. CCO genes were divided into six subfamilies based on the constructed phylogenetic tree, including LbNCEDs, LbCCD1, LbCCD3, LbCCD4, LbCCD7, and LbCCD8. Among them, CCD3 was reported for the first time. The gene structure and motif analysis revealed the conservation of CCO subfamilies. Pseudogene generation and the importance of each subfamily in CCOs were revealed by collinearity analysis. The spatiotemporal transcriptomes of L. barbarum and L. ruthenicum were compared, suggesting that CCD4-1 may dominate carotenoid degradation in goji fruits. Cis-acting elements prediction and environment responsive gene expression analyses indicated that salt-alkali stress and photothermal conditions might influence the expression of CCOs in goji. The results of this study enhance our understanding of the carotenoid degradation pathway, and the functions and responses of CCOs in goji species.
Quantitative Evaluation of the Integrity of Natural Ecosystems and Anthropogenic Impacts in Shennongjia National Park, China
Authors: Huang Zhihao , Cao Jiashuo , Peng Yangjing , Ma Keming , Cui Guofa
Published: May 2023

China launched its pilot national park system in 2015, and Shennongjia National Park has attracted much attention as one of the first batch of pilot national parks. The primary goal of national park construction in China is to effectively protect the integrity and authenticity of nationally important natural ecosystems. Based on the theory of landscape ecology, this study interpreted data from high-resolution remote sensing images and used landscape pattern analysis, spatial correlation analysis, and geographic weighted regression analysis to quantitatively evaluate the integrity of natural ecosystems at the landscape scale. A more accurate and operable calculation method was proposed to analyze the spatial variation in natural ecosystem integrity and to explore the scope and intensity of the impact of different anthropogenic activities on natural ecosystem integrity. The results showed that: (1) from the perspective of the spatial distribution patterns of natural ecosystem integrity, the degree of natural ecosystem integrity of Shennongjia National Park was generally high, with an integrity index value of 96.06, and the proportion of high-integrity areas accounted for 72.38%. The integrity index value of the strict protection zone was 98.83, and the proportion of the low-integrity index area only accounted for 0.01% of the strict protection zone, which was mainly distributed in the main urban areas of the nearby townships and along the highways in the national park, as well as in other areas with intensive anthropogenic activities. (2) From the perspective of the degree of impact of anthropogenic activities on natural ecosystem integrity, population density (0.3344), traffic accessibility (0.2389), traditional utilization activities (0.1101), and industrial and mining activities (0.0095) were, in descending order, the most impactful, and there was no significant correlation between ecotourism activities and natural ecosystem integrity. (3) From the perspective of the impact range of anthropogenic activities on natural ecosystem integrity, traditional utilization activities had the largest impact, accounting for 19.71% of the total area of the national park. The area affected by population density accounted for 1.52%. Industrial and mining activities had an influence of 4.75%, and the area affected by traffic accessibility accounted for 9.28%. Through conducting quantitative research into the integrity of natural ecosystems in Shennongjia National Park, this study provides a new research paradigm for the conservation of natural ecosystems and for the sustainable development of resources in protected areas, which is of great significance for the sensible development of national park conservation and management.
Optimal Fermentation of Artemisia annua Residues and Its Effects on Production Performance of Laying Hens
Authors: Yi Siyu , He Fumeng , Azad Md. Abul Kalam , Zhu Qian , Zhang Minghui , Xu Xiaojie , Cui Yadong , Lan Wei , Li Fenglan , Kong Xiangfeng
Published: May 2023

Artemisia annua residue (ARR) is a pharmaceutical by-product produced after the extraction of artemisinin; it is rich in protein, crude fat, vitamins, trace elements, and bioactive compounds and contains negligible anti-nutritional factors. The present study aimed to optimize the fermentation conditions of ARR, evaluate the compound and microbial compositions of fermented AAR, and explore its effects on the production performance of laying hens. A total of 288 Xinyang black-feather laying hens were randomly allocated into four treatments for 30 days, including a control group (basal diet) and a basal diet supplemented with 1%, 2%, and 4% fermented AAR, respectively. The results showed that the optimized fermentation conditions of AAR were 80% moisture content, 3% inoculation quantity, 34 °C fermentation for 6 days, initial pH at 8, and 60 mesh (sieving). The compounds of 2-furyl-5-methyl furan, deoxyartemisinin, phytol, n-hexadecanoic acid, aromandendrene, and calarene had higher contents (average 6.86%) in the fermented AAR. The bacteria of Proteobacteria and Firmicutes (average 45.18%) were the most abundant phyla, and Acinetobacter, Bacillus, and Brevundimonas (average 15.87%) were the most abundant genera in the fermented AAR. The fungi of Phragmoplastophyta, Vertebrata, and Ascomycota (average 30.13%) were the most abundant phyla, and Magnoliophyta, Mammalia, Wickerhamomyces-Candida_clade, and Aspergillus were the most abundant genera (average 21.12%) in the fermented AAR. Furthermore, dietary supplementation of fermented AAR increased the average daily feed intake (ADFI), egg weight, and albumen height. Dietary supplementation of 2% and 4% fermented AAR increased the laying rate, while 2% fermented AAR increased the Haugh unit and decreased the feed-to-egg ratio. Collectively, it is concluded that fermented AAR has the potential to become a phytogenic feed additive, and dietary supplementation of 2% fermented AAR had better effects on the production performance of laying hens.
Probiotic Properties of Lactic Acid Bacteria Isolated from the Spontaneously Fermented Soybean Foods of the Eastern Himalayas
Authors: Kharnaior Pynhunlang , Tamang Jyoti Prakash
Published: May 2023

Spontaneously fermented soybean foods with sticky-textured and umami-flavor are popular delicacies of multi-ethnic communities of the Eastern Himalayas. Even though species of Bacillus have been reported earlier as pre-dominant bacteria, we hypothesized that some lactic acid bacteria (LAB) present in these unique soy-based foods may exhibit probiotic properties. Hence, the present study is aimed to evaluate some probiotic attributes of LAB. A total of 352 bacterial isolates from spontaneously fermented soybean foods of the Eastern Himalayas viz., kinema, grep-chhurpi, peha, peron namsing and peruñyaan were preliminarily screened for survival in low pH, bile salt tolerance, and cell surface hydrophobicity. Finally, eight probiotic LAB were selected and identified, based on the 16S rRNA gene sequencing, as Pediococcus acidilactici Ki20 and Enterococcus faecium Kn19 (isolated from kinema), E. faecalis Gc21 (grep-chhurpi), P. acidilactici Ph32 (peha), E. faecium Pn11 and E. faecalis Pn37 (peron namsing), E. lactis Py03, and E. lactis Py08 (peruñyaan). In vitro probiotic attributes, E. faecium Kn19 (73.67 ± 1.05) and P. acidilactici Ph32 (79.71 ± 0.13%) recorded higher survival ability in acid and bile salt test, respectively. Furthermore, attachment ability of isolates to hydrocarbons showed ≥80% adhesion property with E. faecalis Gc21 (90.50 ± 10.14%) marked the highest degree of hydrophobicity, and P. acidilactici Ki20 showed the higher auto-aggregation and co-aggregation property. LAB strains were able to produce antibacterial activity against pathogenic bacteria. Genetic screening revealed the presence of genes responsible for acid tolerance (groEl, clpL), bile salt tolerance (apf, bsh), adhesion (msa, mub1), and bacteriocin producing genes for pediocin (pedA, pedB) and enterocin (entA, entB). The present study highlighted the probiotic potentials of LAB strains isolated from Himalayan naturally fermented soybean foods that may be developed as a starter or co-starter culture for controlled and optimized fermentation of soybeans.
Analysis of Phenolic Acids and Flavonoids in Rabbiteye Blueberry Leaves by UPLC-MS/MS and Preparation of Nanoemulsions and Extracts for Improving Antiaging Effects in Mice
Authors: Yu Hsin-Rong , Chen Bing-Huei
Published: May 2023

Rabbiteye blueberry leaves, a waste produced after harvest of blueberry, are rich in polyphenols. This study aims to analyze phenolic acids and flavonoids in blueberry leaves by UPLC-MS/MS and prepare nanoemulsions for determining anti-aging activity in mice. Overall, 30% ethanol was the most suitable extraction solvent for total phenolic acids and total flavonoids. A total of four phenolic acids and four flavonoids were separated within seven minutes for further identification and quantitation by UPLC-MS/MS in selective reaction monitoring (SRM) mode, with 3-O-caffeoylquinic acid being present in the highest amount (6474.2 μg/g), followed by quercetin-3-O-galactoside (1943.9 μg/g), quercetin-3-O-rutinoside (1036.6 μg/g), quercetin-3-O-glucoside (867.2 μg/g), 5-O-caffeoylquinic acid (815.8 μg/g), kaempferol-3-O-glucoside (309.7 μg/g), 3,5-dicaffeoylquinic acid (195.3 μg/g), and 4,5-dicaffeoylquinic acid (60.8 μg/g). The blueberry nanoemulsion was prepared by using an appropriate ratio of soybean oil, Tween 80, glycerol, ethanol, and water at 1.2%, 8%, 2%, 2%, and 86.8%, respectively, and mixing with dried blueberry extract, with the mean particle size and zeta potential being 16 nm and −54 mV, respectively. A high stability was observed during storage of nanoemulsion for 90 days at 4 °C and heated at 100 °C for 2 h. An animal study revealed that this nanoemulsion could elevate dopamine content in mice brain as well as superoxide dismutase, glutathione peroxidase, and catalase activities in mice liver while reducing the contents of malondialdehyde and protein carbonyl in mice brains. Collectively, the high-dose nanoemulsion possessed the highest efficiency in improving mice aging with a promising potential for development into a health food.
Study on the Trend in Microbial Changes during the Fermentation of Black Tea and Its Effect on the Quality
Authors: Liu Changwei , Lin Haiyan , Wang Kuofei , Zhang Zhixu , Huang Jianan , Liu Zhonghua
Published: May 2023

The role of tea endophytes in black tea fermentation and their impact on black tea quality remain unclear. We collected fresh leaves of Bixiangzao and Mingfeng tea and processed them into black tea, while testing the biochemical composition of both the fresh leaves and the black tea. We also used high-throughput techniques, such as 16S rRNA, to analyze the dynamic changes in the microbial community structure and function during black tea processing in order to investigate the influence of dominant microorganisms on the quality of black tea formation. Our results showed that bacteria, such as Chryseobacterium and Sphingomonas, and Pleosporales fungi dominated the entire black tea fermentation process. Predicted functional analysis of the bacterial community indicated that glycolysis-related enzymes, pyruvate dehydrogenase, and tricarboxylic acid cycle-related enzymes were significantly elevated during the fermentation stage. Amino acids, soluble sugars, and tea pigment content also increased considerably during fermentation. Pearson’s correlation analysis revealed that the relative bacterial abundance was closely related to the content of tea polyphenols and catechins. This study provides new insights into the changes in microbial communities during the fermentation of black tea and demonstrates understanding of the basic functional microorganisms involved in black tea processing.
Sociodemographically Stratified Exploration of Pancreatic Cancer Incidence in Younger US Patients: Implication of Cannabis Exposure as a Risk Factor
Authors: Reece Albert Stuart , Hulse Gary Kenneth
Published: May 2023

Introduction. The aetiology for the recent increase in pancreatic cancer incidence (PCI) in the US is unknown. This paper provides an epidemiological investigation of the exponential increase in PCI in young people aged 15–34 years, particularly amongst females, with a focus on the exponential rise amongst African American females, and its relationship to substance use. Methods. National pancreatic cancer data from recent reports. Tobacco, alcohol and daily cannabis use data taken from the annual nationally representative National Survey of Drug Use and Health, response rate = 74%. Results. Amongst the 15–34-year-aged cohort, PCI was found to be significantly more common in females (females: β-est. = 0.1749 p = 0.0005). African American females are noted to have the highest rates of daily cannabis use amongst females in the 26–34 and 35–49-year groups. The relationship between PCI and daily cannabis use was strongly positive across all ethnicities and in both sexes. In African American females, the Pearson correlation between daily cannabis use and PCI was R = 0.8539, p = 0.0051. In an additive multivariable model for each sex and race, cannabis was the only significant term remaining in the final model in the 15–34-year-aged cohort and thus out-performed alcohol as a risk factor. The most significant term in multivariate models was the alcohol:cannabis interaction which was highly significant in all ethnicities from p = 2.50 × 10−7 for Caucasian American females and the highest E-value pair was for Hispanic American females (E-value estimate = 1.26 × 10102 and E-value lower bound 2.20 × 1074). Conclusion. These data show that cannabis fulfills quantitative criteria of causality in all age, sex and ethnicity cohorts, and thus explains both the recent surge in PCI and its ethnocentric predominance. Cannabis interacts powerfully genotoxically and cancerogenically with alcohol, with increases in cannabis use driving the current PCI surge. These results raise the important question as to how much cannabis might be responsible for the modern renaissance in cancer rates amongst younger people.
Quality-by-Design-Assisted Optimization of Carvacrol Oil-Loaded Niosomal Gel for Anti-Inflammatory Efficacy by Topical Route
Authors: Ghazwani Mohammed , Hani Umme , Alam Aftab , Alqarni Mohammed H.
Published: May 2023

Niosomes are multilamellar vesicles that effectively transfer active ingredients into the skin’s layers. To improve the active substance’s penetration across the skin, these carriers are frequently utilized as topical drug delivery systems. Essential oils (EOs) have garnered significant interest in the field of research and development owing to their various pharmacological activities, cost-effectiveness, and simple manufacturing techniques. However, these ingredients undergo degradation and oxidation over time, leading to a loss of functionality. Niosome formulations have been developed to deal with these challenges. The main goal of this work was to create a niosomal gel of carvacrol oil (CVC) to improve its penetration into the skin for anti-inflammatory actions and stability. By changing the ratio of drug, cholesterol and surfactant, various formulations of CVC niosomes were formulated using Box Behnken Design (BBD). A thin-film hydration technique using a rotary evaporator was employed for the development of niosomes. Following optimization, the CVC-loaded niosomes had shown: 180.23 nm, 0.265, −31.70 mV, and 90.61% of vesicle size, PDI, zeta potential, and EE%. An in vitro study on drug release discovered the rates of drug release for CVC-Ns and CVC suspension, which were found to be 70.24 ± 1.21 and 32.87 ± 1.03, respectively. The release of CVC from niosomes best fit the Higuchi model, and the Korsmeyer–Peppas model suggests that the release of the drug followed the non-Fickian diffusion. In a dermatokinetic investigation, niosome gel significantly increased CVC transport in the skin layers when compared to CVC–conventional formulation gel (CVC-CFG). Confocal laser scanning microscopy (CLSM) of rat skin exposed to the rhodamine B-loaded niosome formulation showed a deeper penetration of 25.0 µm compared to the hydroalcoholic rhodamine B solution (5.0 µm). Additionally, the CVC-N gel antioxidant activity was higher than that of free CVC. The formulation coded F4 was selected as the optimized formulation and then gelled with carbopol to improve its topical application. Niosomal gel underwent tests for pH determination, spreadability, texture analysis, and CLSM. Our findings imply that the niosomal gel formulations could represent a potential strategy for the topical delivery of CVC in the treatment of inflammatory disease.
PPIGCF: A Protein–Protein Interaction-Based Gene Correlation Filter for Optimal Gene Selection
Authors: Pati Soumen Kumar , Gupta Manan Kumar , Banerjee Ayan , Mallik Saurav , Zhao Zhongming
Published: May 2023

Biological data at the omics level are highly complex, requiring powerful computational approaches to identifying significant intrinsic characteristics to further search for informative markers involved in the studied phenotype. In this paper, we propose a novel dimension reduction technique, protein–protein interaction-based gene correlation filtration (PPIGCF), which builds on gene ontology (GO) and protein–protein interaction (PPI) structures to analyze microarray gene expression data. PPIGCF first extracts the gene symbols with their expression from the experimental dataset, and then, classifies them based on GO biological process (BP) and cellular component (CC) annotations. Every classification group inherits all the information on its CCs, corresponding to the BPs, to establish a PPI network. Then, the gene correlation filter (regarding gene rank and the proposed correlation coefficient) is computed on every network and eradicates a few weakly correlated genes connected with their corresponding networks. PPIGCF finds the information content (IC) of the other genes related to the PPI network and takes only the genes with the highest IC values. The satisfactory results of PPIGCF are used to prioritize significant genes. We performed a comparison with current methods to demonstrate our technique’s efficiency. From the experiment, it can be concluded that PPIGCF needs fewer genes to reach reasonable accuracy (~99%) for cancer classification. This paper reduces the computational complexity and enhances the time complexity of biomarker discovery from datasets.
The Effects of a Beauty Program on Self-Perception of Aging and Depression among Community-Dwelling Older Adults in an Agricultural Area in Taiwan
Authors: Wu Ya-Ling , Chao Shan-Ru
Published: May 2023

This study aimed to investigate the effects of a beauty program on the self-perception of aging and depression among the community-dwelling older adults in an agricultural area in Taiwan. Twenty-nine older adults aged 65 and above in one agricultural community care center completed the program. Based on cosmetic therapy, the beauty program consisted of 13 sessions focused on facial skin care, make-up application, and massage with essential oils. Each 90 min session of the program was conducted in groups once a week for 13 weeks. This study applied the mixed methods approach, and data were gathered through questionnaire surveys, interviews, and observation. Before and after the beauty program, the elderly individuals’ self-perceptions of aging and depression were assessed using the Attitudes towards Old People Scale (ATOPS) and Taiwanese Depression Questionnaire (TDQ), respectively. The participants’ ATOPS scores after the program were significantly higher than those examined before the program (p < 0.001), and their TDQ scores were significantly lower than those before the program (p < 0.001). Additionally, the participants’ body images were improved, the participants disrupted their stereotypes about makeup, and they were willing to gradually maintain their appearance. Overall, the beauty program was effective for enhancing the self-perceptions of aging and reducing depression in older adults in rural Taiwan. Further research with a larger population of older individuals, male older adults, or frail older adults is needed to examine the specific effects of the beauty program.
Editorial to the Special Issue “The Effect of Exercise Intensity and Psychological Intervention on Musculoskeletal Disorders”
Authors: Sánchez-Romero Eleuterio A. , de-Pedro Miguel , Fernández-Carnero Josué
Published: May 2023

Musculoskeletal disorders are responsible for the most prevalent form of pain, and necessitate a comprehensive approach to rehabilitation [...]
Shifts in Diabetes Health Literacy Policy and Practice in Australia—Promoting Organisational Health Literacy
Authors: Murfet Giuliana O. , Lin Shanshan , Ridd Jan C. , Cremer Gunhild H. , Davidson Susan , Muscat Danielle M.
Published: May 2023

Improving organisational health literacy ensures people can navigate, understand and use essential health information and services. However, systematic reviews have identified limited evidence for practical approaches to implementing such organisational change, particularly at a national level. This study aimed to (a) investigate the approach taken by an Australian national diabetes organisation—Diabetes Australia, as the administrator of the National Diabetes Services Scheme (NDSS)—to improve organisational health literacy over a 15-year-period and (b) examine the impact of organisational changes on the health literacy demands of health information. We performed an environmental scan, examining the websites of the NDSS, Diabetes Australia and the Australian government for reports and position statements describing organisational health literacy policies and practices between 2006 and 2021. The Patient Education Materials Assessment Tool (PEMAT) was applied to consecutively published NDSS diabetes self-care fact sheets (n = 20) to assess changes in the health literacy demands (understandability and actionability) of these fact sheets over the same period. We identified nine policies resulting in 24 health literacy practice changes or projects between 2006 and 2021, applied using a streamlined incremental approach and group reflexivity. The incremental approach focused on (1) increasing audience reach, (2) consistency and branding, (3) person-centred language and (4) the understandability and actionability of health information. The PEMAT scores of fact sheets improved between 2006 and 2021 for understandability (53% to 79%) and actionability (43% to 82%). Diabetes Australia’s information development process leveraging national policies, employing an incremental approach and group reflexivity has improved the health literacy demands of diabetes information and serves as a template for other organisations seeking to improve their organisational health literacy.
Healthy Ageing Should Be a Key Component of Ageing in Place: Case Study from Hong Kong
Authors: Woo Jean , Ko Rina , Yu Ruby , Chan Stacey , Lo Regina , Mo Kar Him
Published: May 2023

As part of a knowledge-transfer project consisting of a series of three talks on the topic of healthy ageing and ageing in place, we explored what participants (older adults, students, the general public, as well as professionals in architecture, urban planning and property management) consider to be key requirements for ageing in place and healthy ageing. Feedback is captured using survey questionnaires and a post-talk discussion group. Safety, a comfortable and spacious environment, age-friendly facilities and meeting the needs of older adults, the availability of caring support and home maintenance services were the most frequently mentioned desirable features of ageing in place. Future models for different types of support for ageing in place may be explored by management companies working with the residents themselves, to develop a sustainable business model.
New Frontiers in Autoimmune Diagnostics: A Systematic Review on Saliva Testing
Authors: Foddai Silvia Grazietta , Radin Massimo , Barinotti Alice , Cecchi Irene , Rubini Elena , Arbrile Marta , Mantello Ester , Menegatti Elisa , Roccatello Dario , Sciascia Savino
Published: May 2023

(1) Background: Immunological laboratory testing is known to be complex, and it is usually performed in tertiary referral centers. Many criticalities affect diagnostic immunological testing, such as limited availability, the need for specifically trained laboratory staff, and potential difficulties in collecting blood samples, especially in the most vulnerable patients, i.e., the elderly and children. For this reason, the identification of a new feasible and reliable methodology for autoantibody detection is urgently needed. (2) Methods: We designed a systematic review to investigate the available literature on the utilization of saliva samples for immunological testing. (3) Results: A total of 170 articles were identified. Eighteen studies met the inclusion criteria, accounting for 1059 patients and 671 controls. The saliva collection method was mostly represented by passive drooling (11/18, 61%), and the most frequently described methodology for antibody detection was ELISA (12/18, 67%). The analysis included 392 patients with rheumatoid arthritis, 161 with systemic lupus erythematosus, 131 with type 1 diabetes mellitus, 116 with primary biliary cholangitis, 100 with pemphigus vulgaris, 50 with bullous pemphigoids, 49 with Sjogren syndrome, 39 with celiac disease, 10 with primary antiphospholipid syndromes, 8 with undifferentiated connective tissue disease, 2 with systemic sclerosis, and 1 with autoimmune thyroiditis. The majority of the reviewed studies involved adequate controls, and saliva testing allowed for a clear distinction of patients (10/12 studies, 83%). More than half of the papers showed a correlation between saliva and serum results (10/18, 55%) for autoantibody detection, with varying rates of correlation, sensitivity, and specificity. Interestingly, many papers showed a correlation between saliva antibody results and clinical manifestations. (4) Conclusions: Saliva testing might represent an appealing alternative to serum-based testing for autoantibody detection, considering the correspondence with serum testing results and the correlation with clinical manifestations. Nonetheless, standardization of sample collection processing, maintenance, and detection methodology has yet to be fully addressed.
Multiplexed Gene Engineering Based on dCas9 and gRNA-tRNA Array Encoded on Single Transcript
Authors: Jiang Chaoqian , Geng Lishuang , Wang Jinpeng , Liang Yingjuan , Guo Xiaochen , Liu Chang , Zhao Yunjing , Jin Junxue , Liu Zhonghua , Mu Yanshuang
Published: May 2023

Simultaneously, multiplexed genome engineering and targeting multiple genomic loci are valuable to elucidating gene interactions and characterizing genetic networks that affect phenotypes. Here, we developed a general CRISPR-based platform to perform four functions and target multiple genome loci encoded in a single transcript. To establish multiple functions for multiple loci targets, we fused four RNA hairpins, MS2, PP7, com and boxB, to stem–loops of gRNA (guide RNA) scaffolds, separately. The RNA-hairpin-binding domains MCP, PCP, Com and λN22 were fused with different functional effectors. These paired combinations of cognate-RNA hairpins and RNA-binding proteins generated the simultaneous, independent regulation of multiple target genes. To ensure that all proteins and RNAs are expressed in one transcript, multiple gRNAs were constructed in a tandemly arrayed tRNA (transfer RNA)-gRNA architecture, and the triplex sequence was cloned between the protein-coding sequences and the tRNA-gRNA array. By leveraging this system, we illustrate the transcriptional activation, transcriptional repression, DNA methylation and DNA demethylation of endogenous targets using up to 16 individual CRISPR gRNAs delivered on a single transcript. This system provides a powerful platform to investigate synthetic biology questions and engineer complex-phenotype medical applications.
Can Pharmacogenetic Variants in TPMT, MTHFR and SLCO1B1 Genes Be Used as Potential Markers of Outcome Prediction in Systemic Sclerosis Patients?
Authors: Jelovac Marina , Kotur Nikola , Ristivojevic Bojan , Pavlovic Djordje , Spasovski Vesna , Damjanov Nemanja , Pavlovic Sonja , Zukic Branka
Published: May 2023

Systemic sclerosis (SSc) is a rare connective tissue disorder with highest morbidity and mortality among rheumatologic diseases. Disease progression is highly heterogeneous between patients, implying a strong need for individualization of therapy. Four pharmacogenetic variants, namely TPMT rs1800460, TPMT rs1142345, MTHFR rs1801133 and SLCO1B1 rs4149056 were tested for association with severe disease outcomes in 102 patients with SSc from Serbia treated either with immunosuppressants azathioprine (AZA) and methotrexate (MTX) or with other types of medications. Genotyping was performed using PCR-RFLP and direct Sanger sequencing. R software was used for statistical analysis and development of polygenic risk score (PRS) model. Association was found between MTHFR rs1801133 and higher risk for elevated systolic pressure in all patients except those prescribed with MTX, and higher risk for kidney insufficiency in patients prescribed with other types of drugs. In patients treated with MTX, variant SLCO1B1 rs4149056 was protective against kidney insufficiency. For patients receiving MTX a trend was shown for having a higher PRS rank and elevated systolic pressure. Our results open a door wide for more extensive research on pharmacogenomics markers in patients with SSc. Altogether, pharmacogenomics markers could predict the outcome of patients with SSc and help in prevention of adverse drug reactions.
Submergence Stress Alters the Expression of Clock Genes and Configures New Zeniths and Expression of Outputs in Brachypodium distachyon
Authors: Medina-Chávez Lucisabel , Camacho Christian , Martínez-Rodríguez Jorge Arturo , Barrera-Figueroa Blanca Estela , Nagel Dawn H. , Juntawong Piyada , Peña-Castro Julián Mario
Published: May 2023

Plant networks of oscillating genes coordinate internal processes with external cues, contributing to increased fitness. We hypothesized that the response to submergence stress may dynamically change during different times of the day. In this work, we determined the transcriptome (RNA sequencing) of the model monocotyledonous plant, Brachypodium distachyon, during a day of submergence stress, low light, and normal growth. Two ecotypes of differential tolerance, Bd21 (sensitive) and Bd21-3 (tolerant), were included. We submerged 15-day-old plants under a long-day diurnal cycle (16 h light/8 h dark) and collected samples after 8 h of submergence at ZT0 (dawn), ZT8 (midday), ZT16 (dusk), ZT20 (midnight), and ZT24 (dawn). Rhythmic processes were enriched both with up- and down-regulated genes, and clustering highlighted that the morning and daytime oscillator components (PRRs) show peak expression in the night, and a decrease in the amplitude of the clock genes (GI, LHY, RVE) was observed. Outputs included photosynthesis-related genes losing their known rhythmic expression. Up-regulated genes included oscillating suppressors of growth, hormone-related genes with new late zeniths (e.g., JAZ1, ZEP), and mitochondrial and carbohydrate signaling genes with shifted zeniths. The results highlighted genes up-regulated in the tolerant ecotype such as METALLOTHONEIN3 and ATPase INHIBITOR FACTOR. Finally, we show by luciferase assays that Arabidopsis thaliana clock genes are also altered by submergence changing their amplitude and phase. This study can guide the research of chronocultural strategies and diurnal-associated tolerance mechanisms.
Genome-Wide Analysis of the BBX Genes in Platanus × acerifolia and Their Relationship with Flowering and/or Dormancy
Authors: Shi Gehui , Ai Kangyu , Yan Xu , Zhou Zheng , Cai Fangfang , Bao Manzhu , Zhang Jiaqi
Published: May 2023

The B-BOX (BBX) gene family is widely distributed in animals and plants and is involved in the regulation of their growth and development. In plants, BBX genes play important roles in hormone signaling, biotic and abiotic stress, light-regulated photomorphogenesis, flowering, shade response, and pigment accumulation. However, there has been no systematic analysis of the BBX family in Platanus × acerifolia. In this study, we identified 39 BBX genes from the P. × acerifolia genome, and used TBtools, MEGA, MEME, NCBI CCD, PLANTCARE and other tools for gene collinearity analysis, phylogenetic analysis, gene structure, conserved domain analysis, and promoter cis-element analysis, and used the qRT-PCR and transcriptome data for analyzing expression pattern of the PaBBX genes. Collinearity analysis indicated segmental duplication was the main driver of the BBX family in P. × acerifolia, and phylogenetic analysis showed that the PaBBX family was divided into five subfamilies: I, II, III, IV and V. Gene structure analysis showed that some PaBBX genes contained super-long introns that may regulate their own expression. Moreover, the promoter of PaBBX genes contained a significant number of cis-acting elements that are associated with plant growth and development, as well as hormone and stress responses. The qRT-PCR results and transcriptome data indicated that certain PaBBX genes exhibited tissue-specific and stage-specific expression patterns, suggesting that these genes may have distinct regulatory roles in P. × acerifolia growth and development. In addition, some PaBBX genes were regularly expressed during the annual growth of P. × acerifolia, corresponding to different stages of flower transition, dormancy, and bud break, indicating that these genes may be involved in the regulation of flowering and/or dormancy of P. × acerifolia. This article provided new ideas for the study of dormancy regulation and annual growth patterns in perennial deciduous plants.
From the Gender Gap to Neuroactive Steroids: Exploring Multiple Cases to Further Understand Neuropathic Pain
Authors: Marinelli Sara , Coccurello Roberto
Published: May 2023

Neuropathic pain (NeuP) is still an intractable form of highly debilitating chronic pain, resulting from a lesion or disease of the somatosensory nervous system [...]
An Elusive Target: Inhibitors of JC Polyomavirus Infection and Their Development as Therapeutics for the Treatment of Progressive Multifocal Leukoencephalopathy
Authors: Kaiserman Jacob , O’Hara Bethany A. , Haley Sheila A. , Atwood Walter J.
Published: May 2023

Progressive multifocal leukoencephalopathy (PML) is a rare demyelinating disease caused by infection with JC Polyomavirus (JCPyV). Despite the identification of the disease and isolation of the causative pathogen over fifty years ago, no antiviral treatments or prophylactic vaccines exist. Disease onset is usually associated with immunosuppression, and current treatment guidelines are limited to restoring immune function. This review summarizes the drugs and small molecules that have been shown to inhibit JCPyV infection and spread. Paying attention to historical developments in the field, we discuss key steps of the virus lifecycle and antivirals known to inhibit each event. We review current obstacles in PML drug discovery, including the difficulties associated with compound penetrance into the central nervous system. We also summarize recent findings in our laboratory regarding the potent anti-JCPyV activity of a novel compound that antagonizes the virus-induced signaling events necessary to establish a productive infection. Understanding the current panel of antiviral compounds will help center the field for future drug discovery efforts.
Exploring BenzylethoxyAryl Urea Scaffolds for Multitarget Immunomodulation Therapies
Authors: Gil-Edo Raquel , Hernández-Ribelles German , Royo Santiago , Thawait Natasha , Serrels Alan , Carda Miguel , Falomir Eva
Published: May 2023

Thirteen benzylethoxyaryl ureas have been synthesized and biologically evaluated as multitarget inhibitors of VEGFR-2 and PD-L1 proteins to overcome resistance phenomena offered by cancer. The antiproliferative activity of these molecules on several tumor cell lines (HT-29 and A549), on the endothelial cell line HMEC-1, on immune cells (Jurkat T) and on the non-tumor cell line HEK-293 has been determined. Selective indexes (SI) have been also determined and compounds bearing p-substituted phenyl urea unit together with a diaryl carbamate exhibited high SI values. Further studies on these selected compounds to determine their potential as small molecule immune potentiators (SMIPs) and as antitumor agents have been performed. From these studies, we have concluded that the designed ureas have good tumor antiangiogenic properties, exhibit good inhibition of CD11b expression, and regulate pathways involved in CD8 T-cell activity. These properties suggest that these compounds could be potentially useful in the development of new cancer immune treatments.
The Synergistic Effect of Interventional Locoregional Treatments and Immunotherapy for the Treatment of Hepatocellular Carcinoma
Authors: Brandi Nicolò , Renzulli Matteo
Published: May 2023

Immunotherapy has remarkably revolutionized the management of advanced HCC and prompted clinical trials, with therapeutic agents being used to selectively target immune cells rather than cancer cells. Currently, there is great interest in the possibility of combining locoregional treatments with immunotherapy for HCC, as this combination is emerging as an effective and synergistic tool for enhancing immunity. On the one hand, immunotherapy could amplify and prolong the antitumoral immune response of locoregional treatments, improving patients’ outcomes and reducing recurrence rates. On the other hand, locoregional therapies have been shown to positively alter the tumor immune microenvironment and could therefore enhance the efficacy of immunotherapy. Despite the encouraging results, many unanswered questions still remain, including which immunotherapy and locoregional treatment can guarantee the best survival and clinical outcomes; the most effective timing and sequence to obtain the most effective therapeutic response; and which biological and/or genetic biomarkers can be used to identify patients likely to benefit from this combined approach. Based on the current reported evidence and ongoing trials, the present review summarizes the current application of immunotherapy in combination with locoregional therapies for the treatment of HCC, and provides a critical evaluation of the current status and future directions.
Transcriptome and Metabolome Reveal Distinct Sugar Accumulation Pattern between PCNA and PCA Mature Persimmon Fruit
Authors: Han Weijuan , Wang Yiru , Li Huawei , Diao Songfeng , Suo Yujing , Li Taishan , Sun Peng , Li Fangdong , Fu Jianmin
Published: May 2023

Persimmon (Diospyros kaki) fruit have significant variation between pollination-constant non-astringent (PCNA) and pollination-constant astringent (PCA) persimmons. The astringency type affects not only the soluble tannin concentration but also the accumulation of individual sugars. Thus, we comprehensively investigate the gene expression and metabolite profiles of individual sugars to resolve the formation of flavor differences in PCNA and PCA persimmon fruit. The results showed that soluble sugar, starch content, sucrose synthase, and sucrose invertase were significantly different between PCNA and PCA persimmon fruit. The sucrose and starch metabolism pathway was considerably enriched, and six sugar metabolites involving this pathway were significantly differentially accumulated. In addition, the expression patterns of diferentially expressed genes (such as bglX, eglC, Cel, TPS, SUS, and TREH genes) were significantly correlated with the content of deferentially accumulated metabolites (such as starch, sucrose, and trehalose) in the sucrose and starch metabolism pathway. These results indicated that the sucrose and starch metabolism pathway maintained a central position of sugar metabolism between PCNA and PCA persimmon fruit. Our results provide a theoretical basis for exploring functional genes related to sugar metabolism and provide useful resources for future studies on the flavor differences between PCNA and PCA persimmon fruit.
A Case for Below-Ground Dispersal? Insights into the Biology, Ecology and Conservation of Blind Cave Spiders in the Genus Troglodiplura (Mygalomorphae: Anamidae)
Authors: Marsh Jessica R. , Milner Steven J. , Shaw Matthew , Stempel Andrew J. , Harvey Mark S. , Rix Michael G.
Published: May 2023

Previously described from only fragments of exoskeleton and juvenile specimens, the cave spider genus Troglodiplura (Araneae: Anamidae), endemic to the Nullarbor Plain, is the only troglomorphic member of the infraorder Mygalomorphae recorded from Australia. We investigated the distribution of Troglodiplura in South Australia, collecting and observing the first (intact) mature specimens, widening the number of caves it has been recorded in, and documenting threats to conservation. Phylogenetic analyses support the placement of Troglodiplura as an independent lineage within the subfamily Anaminae (the ‘Troglodiplura group’) and provide unequivocal evidence that populations from apparently isolated cave systems are conspecifics of T. beirutpakbarai Harvey & Rix, 2020, with extremely low or negligible inter-population mitochondrial divergences. This is intriguing evidence for recent or contemporary subterranean dispersal of these large, troglomorphic spiders. Observations of adults and juvenile spiders taken in the natural cave environment, and supported by observations in captivity, revealed the use of crevices within caves as shelters, but no evidence of silk use for burrow construction, contrasting with the typical burrowing behaviours seen in other Anamidae. We identify a range of threats posed to the species and to the fragile cave ecosystem, and provide recommendations for further research to better define the distribution of vulnerable taxa within caves and identify actions needed to protect them.
Ten-Year Trends in Psychotropic Prescribing and Polypharmacy in Australian General Practice Patients with and without Dementia
Authors: Bezabhe Woldesellassie , Radford Jan , Salahudeen Mohammed , Bindoff Ivan , Ling Tristan , Gee Peter , Wimmer Barbara , Peterson Gregory
Published: May 2023

Objective: Little research has evaluated trends in psychotropic prescribing and polypharmacy in primary care patients, especially those with dementia. We sought to examine this in Australia from 2011 to 2020 using the primary care dataset, MedicineInsight. Methods: Ten consecutive serial cross-sectional analyses were performed to evaluate the proportion of patients aged 65 years or more, with a recorded diagnosis of dementia, who were prescribed psychotropic medications within the first six months of each year from 2011 to 2020. This proportion was compared with propensity score-matched control patients without dementia. Results: Before matching, 24,701 patients (59.2% females) with, and 72,105 patients (59.2% females) without, a recorded diagnosis of dementia were included. In 2011, 42% (95% confidence interval [CI] 40.5–43.5%) of patients in the dementia group had at least one recorded prescription of a psychotropic medication, which declined to 34.2% (95% CI 33.3–35.1%; p for trend < 0.001) by 2020. However, it remained unchanged for matched controls (36% [95% CI 34.6–37.5%] in 2011 and 36.7% [95% CI 35.7–37.6%] in 2020). The greatest decline in the dementia groups by medication class was for antipsychotics (from 15.9% [95% CI 14.8–17.0%] to 8.8% [95% CI 8.2–9.4%]; p for trend < 0.001). During this period, the prevalence of psychotropic polypharmacy (use of two or more individual psychotropics) also decreased from 21.7% (95% CI 20.5–22.9%) to 18.1% (95% CI 17.4–18.9%) in the dementia groups, and slightly increased from 15.2% (95% CI 14.1–16.3%) to 16.6% (95% CI 15.9–17.3%) in the matched controls. Conclusions: The decline in psychotropic prescribing, particularly antipsychotics, in Australian primary care patients with dementia is encouraging. However, psychotropic polypharmacy still occurred in almost one in five patients with dementia at the end of the study period. Programs focused on encouraging further reductions in the use of multiple psychotropic drugs in patients with dementia are recommended, particularly in rural and remote regions.
History and Future of HER2-Targeted Therapy for Advanced Gastric Cancer
Authors: Ariga Shin
Published: May 2023

Human epidermal growth factor receptor 2 (HER2) is a receptor tyrosine kinase that belongs to the human epidermal growth factor receptor family. It is overexpressed/amplified in approximately 20% of gastric or gastroesophageal junction cancers. HER2 is being developed as a therapeutic target in a variety of cancers, and several agents have been shown to be effective in breast cancer. The development of HER2-targeted therapy for gastric cancer successfully began with trastuzumab. However, while effective in breast cancer, the successive anti-HER2 agents lapatinib, T-DM1, and pertuzumab failed to demonstrate benefits regarding survival in gastric cancer compared with the existing standard therapies. Intrinsic differences lie between gastric and breast cancer in terms of HER2-positive tumor biology, which may make development difficult. Recently, a novel anti-HER2 agent, trastuzumab deruxtecan, was introduced, and the development of HER2-positive gastric cancer agents has been moving to the next stage. This review summarizes the current HER2-targeted therapy for gastric or gastroesophageal cancer in chronological order and describes the promising future of HER2-targeted therapy.
Current Role of Intracoronary Imaging for Implementing Risk Stratification and Tailoring Culprit Lesion Treatment: A Narrative Review
Authors: Fabris Enrico , Kedhi Elvin , Verdoia Monica , Ielasi Alfonso , Tespili Maurizio , Guagliumi Giulio , De Luca Giuseppe
Published: May 2023

Our understanding of the pathophysiology of acute coronary syndrome and of the vascular biology of coronary atherosclerosis has made enormous progress with the implementation of intravascular imaging. Intravascular imaging contributes to overcoming the known limitations of coronary angiography and allows for the in vivo discrimination of plaque morphology giving insight into the underlying pathology of the disease process. The possibility of using intracoronary imaging to characterize lesion morphologies and correlate them with clinical presentations may influence the treatment of patients and improve risk stratification, offering the opportunity for tailored management. This review examines the current role of intravascular imaging and describes how intracoronary imaging represents a valuable tool for modern interventional cardiology in order to improve diagnostic accuracy and offer a tailored approach to the treatment of patients with coronary artery disease, especially in the acute setting.