bluuebunny/crossref_metadata_2025 huggingface dataset:
Dual-Crosslinked Alginate-Based Hydrogels with Tunable Mechanical Properties for Cultured Meat
Authors: Tahir Irfan , Floreani Rachael
Published: September 2022

Cultured meat refers to the production of animal tissue by utilizing the same techniques as tissue engineering through cell culture. Various biomaterials have been designed to serve as in vitro supports for cell viability, growth, and migration. In this study, visible light and dual-crosslinked alginate hydrogels were designed to enable control of the physical and mechanical properties needed for the fabrication of cultured meat scaffolds. We hypothesized that a difference in hydrogel stiffness would influence cell behavior, indicating the efficacy of our processing methods to benefit the cultured meat field. Herein, we synthesized and created: (1) methacrylated alginate (AlgMA) to enable covalent crosslinking via visible light exposure, (2) Methacrylated alginate and arginyl-glycyl-aspartic acid RGD conjugates (AlgMA-RGD), using carbodiimide chemistries to provide cell-binding sites on the material, and (3) designer hydrogels incorporating different crosslinking techniques. The material and mechanical properties were evaluated to determine the structural integrity of the hydrogels, and in vitro cell assays were conducted to verify cytocompatibility and cell adhesion. Gelation, swell ratio, and weight loss calculations revealed longer gelation times for the AlgMA scaffolds and similar physical properties for all hydrogel groups. We showed that by adjusting the polymer concentration and the crosslinking methodology, the scaffold’s mechanical properties can be controlled and optimized within physiological ranges. Incorporating dual crosslinking significantly increased the compressive moduli of the AlgMA hydrogels, compared to visible-light crosslinking alone. Moreover, the muscle satellite cells responded favorably to the AlgMA scaffolds, with clear differences in cell density when cultured on materials with significantly different mechanical properties. Our results indicate the usefulness of the dual-crosslinking alginate hydrogel system to support in vitro meat growth.
Changes in the Primary Metabolites of ‘Fengtang’ Plums during Storage Detected by Widely Targeted Metabolomics
Authors: Lin Xin , Huang Shian , Zhang Qin , Zhu Shouliang , Dong Xiaoqing
Published: September 2022

Plums are one of the most popular stone fruits worldwide owing to their high nutritional value. After harvest, plum fruit quality and flavor change during storage; however, little is known about the changes in metabolites during this period. A comprehensive comparison of primary metabolites in ‘Fengtang’ plum fruits during storage is performed using widely targeted primary metabolomics. A total of 272 primary metabolites were identified by means of ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS) in the plums at different storage periods. There was a significant increase in the relative amounts of twenty-eight lipids, twenty amino acids and their derivatives, thirteen organic acids, ten saccharides and alcohols, six nucleotides and their derivatives, and two vitamins. A Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differential metabolites revealed that glucosinolate biosynthesis, starch and sucrose metabolism, ascorbate and aldarate metabolism, lysine degradation, and other metabolic pathways were significantly enriched; therefore, changes in these metabolic pathways may be key to the quality and flavor change in ‘Fengtang’ plum fruits during storage. Our results provide a theoretical foundation and technical support to evaluate ‘Fengtang’ plum fruit quality.
Single-Cell Proteins Obtained by Circular Economy Intended as a Feed Ingredient in Aquaculture
Authors: Pereira Antia G. , Fraga-Corral Maria , Garcia-Oliveira Paula , Otero Paz , Soria-Lopez Anton , Cassani Lucia , Cao Hui , Xiao Jianbo , Prieto Miguel A. , Simal-Gandara Jesus
Published: September 2022

The constant increment in the world’s population leads to a parallel increase in the demand for food. This situation gives place the need for urgent development of alternative and sustainable resources to satisfy this nutritional requirement. Human nutrition is currently based on fisheries, which accounts for 50% of the fish production for human consumption, but also on agriculture, livestock, and aquaculture. Among them, aquaculture has been pointed out as a promising source of animal protein that can provide the population with high-quality protein food. This productive model has also gained attention due to its fast development. However, several aquaculture species require considerable amounts of fish protein to reach optimal growth rates, which represents its main drawback. Aquaculture needs to become sustainable using renewable source of nutrients with high contents of proteins to ensure properly fed animals. To achieve this goal, different approaches have been considered. In this sense, single-cell protein (SCP) products are a promising solution to replace fish protein from fishmeal. SCP flours based on microbes or algae biomass can be sustainably obtained. These microorganisms can be cultured by using residues supplied by other industries such as agriculture, food, or urban areas. Hence, the application of SCP for developing innovative fish meal offers a double solution by reducing the management of residues and by providing a sustainable source of proteins to aquaculture. However, the use of SCP as aquaculture feed also has some limitations, such as problems of digestibility, presence of toxins, or difficulty to scale-up the production process. In this work, we review the potential sources of SCP, their respective production processes, and their implementation in circular economy strategies, through the revalorization and exploitation of different residues for aquaculture feeding purposes. The data analyzed show the positive effects of SCP inclusion in diets and point to SCP meals as a sustainable feed system. However, new processes need to be exploited to improve yield. In that direction, the circular economy is a potential alternative to produce SCP at any time of the year and from various cost-free substrates, almost without a negative impact.
A Review on Nutrients, Phytochemicals, and Health Benefits of Green Seaweed, Caulerpa lentillifera
Authors: Syakilla Nur , George Ramlah , Chye Fook Yee , Pindi Wolyna , Mantihal Sylvester , Wahab Noorakmar Ab , Fadzwi Fazlini Mohd , Gu Philip Huanqing , Matanjun Patricia
Published: September 2022

Caulerpa lentillifera is a type of green seaweed widely consumed as a fresh vegetable, specifically in Southeast Asia. Interestingly, this green seaweed has recently gained popularity in the food sector. Over the last two decades, many studies have reported that C. lentillifera is rich in polyunsaturated fatty acids, minerals, vitamins, and bioactive compounds that contribute many health benefits. On the other hand, there is currently hardly any article dedicated specifically to C. lentillifera regarding nutritional composition and recent advancements in its potential health benefits. Hence, this study will summarise the findings on the nutritional content of C. lentillifera and compile recently discovered beneficial properties throughout the past decade. From the data compiled in this review paper, it can be concluded that the nutrient and phytochemical profile of C. lentillifera differs from one region to another depending on various external factors. As a result, this paper will offer researchers the groundwork to develop food products based on C. lentillifera. The authors of this paper are hopeful that a more systematic review could be done in the future as currently, existing data is still scarce.
Preparation and Characterization of Aronia melanocarpa/Gellan Gum/Pea Protein/Chitosan Bilayer Films
Authors: Chen Xuanhong , Ji Wei , Nan Xijun , Wang Honglei , Li Jiayi , Dong Leichao , Sheng Guihua , Zhou Quancheng
Published: September 2022

In this study, pH-sensitive bilayer hydrogel films with different AM contents (0.00%, 0.50%, 1.00%, 1.50%, 2.00% and 2.50%) were constructed. The films took AM/GG hydrogel as the inner layer structure and a pea protein (PP)/chitosan (CS) composite system as the outer structure. Film formation and the effect of AM were clarified through the detection and analysis of mechanical properties, microstructure, pH sensitivity and fresh-keeping ability. Results showed that AM exhibited good compatibility with each substance in the composite film, which were evenly dispersed in the system. The addition of AM significantly improved the water content, tensile strength, elongation at break, puncture resistance, oil resistance and water resistance of the composite films. The antioxidant activity, pH sensitivity and fresh-keeping effect of the composite film on fresh pork were remarkably enhanced. Moreover, it was found that the composite film containing AM effectively inhibited the production of total volatile base nitrogen (TVN) in fresh pork and significantly reduced the weight loss of fresh pork due to water loss during storage. Therefore, the functional properties revealed that AM was more positive to the comprehensive performance of films, and the AM-GG/PP-CS bilayer film containing AM exhibited strong potential for use in food preservation and packaging as a food freshness indicator to test food quality changes in storage.
Properties of a Novel Salmonella Phage L66 and Its Application Based on Electrochemical Sensor-Combined AuNPs to Detect Salmonella
Authors: Li Changbin , Wang Yuanshang , Wang Jia , Wang Xiaohong
Published: September 2022

Salmonella is widespread in nature and poses a significant threat to human health and safety. Phage is considered as a new tool for the control of food-borne pathogens. In this study, Salmonella phage L66 (phage L66) was isolated from sewage by using Salmonella Typhimurium ATCC 14028 as the host bacterium, and its basic properties were obtained by biological and bioinformatics analysis. Phage L66 had a broad host spectrum, with an optimal infection complex of 0.1 and an optimal adsorption rate of 90.06%. It also exhibited thermal stability between 30 °C~60 °C and pH stability pH from 3 to 12, and the average lysis amount was 46 PFU/cell. The genome sequence analysis showed that the genome length of phage L66 was 157,675 bp and the average GC content was 46.13%. It was predicted to contain 209 genes, 97 of which were annotated with known functions based on the evolutionary analysis, and phage L66 was attributed to the Kuttervirus genus. Subsequently, an electrochemical sensor using phage L66 as a recognition factor was developed and the working electrode GDE-AuNPs-MPA-Phage L66 was prepared by layer-by-layer assembly for the detection of Salmonella. The slope of the impedance was 0.9985 within the scope from 20 to 2 × 107 CFU/mL of bacterial concentration. The minimum detection limit of the method was 13 CFU/mL, and the average spiked recovery rate was 102.3% with a relative standard deviation of 5.16%. The specificity and stability of this sensor were excellent, and it can be applied for the rapid detection of Salmonella in various foods. It provides a phage-based electrochemical biosensor for the detection of pathogenic bacteria.
New Adaptive Finite-Time Cluster Synchronization of Neutral-Type Complex-Valued Coupled Neural Networks with Mixed Time Delays
Authors: Boonsatit Nattakan , Rajendran Santhakumari , Lim Chee Peng , Jirawattanapanit Anuwat , Mohandas Praneesh
Published: September 2022

The issue of adaptive finite-time cluster synchronization corresponding to neutral-type coupled complex-valued neural networks with mixed delays is examined in this research. A neutral-type coupled complex-valued neural network with mixed delays is more general than that of a traditional neural network, since it considers distributed delays, state delays and coupling delays. In this research, a new adaptive control technique is developed to synchronize neutral-type coupled complex-valued neural networks with mixed delays in finite time. To stabilize the resulting closed-loop system, the Lyapunov stability argument is leveraged to infer the necessary requirements on the control factors. The effectiveness of the proposed method is illustrated through simulation studies.
Quality of Life in Community-Dwelling Older People with Functional and Nutritional Impairment and Depressive Symptoms: A Comparative Cross-Sectional Study in Brazil and Portugal
Authors: de Miranda Jéssica Maria Arouca , Viana Dalyanna Mildred de Oliveira , dos Santos Anderson Antônio Lima , Costa Áquila Filêmon de Andrade , Dantas Bruno Araújo da Silva , de Miranda Francisco Arnoldo Nunes , Mendes Felismina Rosa Parreira , Torres Gilson de Vasconcelos
Published: September 2022

Background: The interaction of quality of life (QoL) with functionality, nutrition and depression has been studied, but few studies have compared different realities. Our objective was to compare the associations of QoL with impaired functionality, nutritional status and depressive symptoms among older people patients treated in primary health care (PHC) in Brazil and Portugal. Methods: Cross-sectional, comparative study was conducted with primary data from PHC services in Brazil and Portugal with users over 65 years old. Participants’ scores were classified as “impaired” and “preserved” for QoL, functional decline, nutrition and depression. We used Pearson’s chi-square test, Fisher’s exact test and the Mann–Whitney U test. Results: Our sample had a total of 150 PHC users. We found lower QoL scores in Brazil, which were associated with the risk of functional decline for the domains Physical Functioning, General Health Perceptions, Mental Health dimensions and Physical Health. Nutritional impairment in the group from Portugal included the domains of Vitality and Social Role Functioning. For depressive impairment, Portugal showed an association with the domains Mental Health, Vitality and Social Role Functioning. Conclusions: QoL was associated with functional and nutritional impairment and depressive symptoms, highlighting physical, mental and social characteristics related to the perception of well-being.
Measurement of Skin Thickness Using Ultrasonography to Test the Usefulness of Elastic Compression Stockings for Leg Edema in Pregnant Women
Authors: Banba Airi , Koshiyama Masafumi , Watanabe Yumiko , Makino Koji , Ikuta Eri , Yanagisawa Nami , Ono Ayumi , Nakagawa Miwa , Seki Keiko , Sakamoto Shin-ichi , Hara Yoko , Nakajima Akira
Published: September 2022

Background: One of the most common treatments for leg edema during pregnancy is the use of compression stockings. The purpose of this study was to evaluate the objective effectiveness in pregnant women, by measuring the changes of skin thickness using ultrasonography. Methods: Pregnant women were diagnosed with leg edema using the pitting edema method at 36 weeks of gestation. Twenty-four pregnant women (48 legs) with leg edema spent time without wearing elastic stockings at 36–37 weeks of gestation. Then, they wore elastic stockings for one week at 37–38 weeks of gestation. We measured the grade of edema (from 0 to 3) and the skin thickness of the lower leg by portable ultrasonography at 36, 37, and 38 weeks of gestation (a before-and-after study). Results: In 24 pregnant women, thigh edema was not detected in any of the 48 legs before or after the use of elastic stockings. All 48 legs in 24 pregnant women had physiological lower leg edema, but not thigh edema. The average grade of pitting edema in each lower leg significantly decreased after using the stockings (36 weeks, 1.77 ± 0.85; 37 weeks, 1.79 ± 0.77; 38 weeks, 1.04 ± 0.74, p < 0.0001). In addition, the skin thickness of the lower legs was significantly decreased after the use of elastic stockings (36 weeks, 7.47 ± 2.45 mm; 37 weeks, 7.93 ± 2.83 mm; 38 weeks, 7.15 ± 2.35 mm, p < 0.0001). Conclusions: The wearing of elastic compression stockings on the lower legs is objectively effective for improving leg edema in pregnant women.
The Involvement of Past Traumatic Life Events in the Development of Postpartum PTSD after Cesarean Delivery
Authors: Orovou Eirini , Dagla Maria , Eskitzis Panagiotis , Savvidis Georgios S. , Rigas Nikolaos , Papatrechas Alexandros , Sarella Angeliki , Arampatzi Christiana , Antoniou Evangelia
Published: September 2022

Background: Although childbirth is considered a natural process, a high percentage of postpartum women consider it traumatic. Any previous traumatic event in a woman’s life can be revived through a traumatic birth experience, especially after a complicated vaginal delivery or cesarean delivery. The purpose of this study was to clarify the relationship between previous traumatic life events and posttraumatic stress disorder (PTSD) in postpartum women after cesarean section and which specific events exerted the greatest influence. Methods: A sample of 469 women who had undergone cesarean sections at a Greek university hospital consented to participate in this prospective study. Data from a medical/demographic questionnaire, life events checklist, perinatal stressor criterion A, and posttraumatic stress checklist were used to evaluate past traumatic life events and diagnose postpartum posttraumatic stress. Results: Out of 469 women, 25.97% had PTSD and 11.5% a PTSD profile, while 2.7% had PTSD and 2.7% a PTSD profile. Also, it appeared that only specific direct exposure to a traumatic event and/or witnessing one were predictors of postpartum PTSD. Conclusions: This survey identified specific traumatic life events, psychiatric history, stressor perinatal criterion A, preterm birth, and emergency cesarean section as risk factors for the development of PTSD or a PTSD profile in women after cesarean delivery.
Environmental Pressures at Dirre Sheikh Hussein Sanctuary
Authors: Tola Habtamu Gizawu , Brimblecombe Peter
Published: September 2022

Dirre Sheikh Hussein is a religious complex in Ethiopia that dates from the 12th century or earlier. An important focus for pilgrimage, it lies in an isolated part of Oromia State on poorly drained semi-arid lowlands. Significant architecturally and culturally, recent expansion and refurbishment of the major buildings led to structural issues, though these are now largely overcome through consolidation of the prayer hall roof, laying a stone walkway, digging drainage ditches etc. Current deterioration of the buildings can be seen as: (i) fading of colours, (ii) cracks in walls, roofs, and festive places, (iii) deposits of sand around walls and (iv) staining and drainage marks on the white painted surfaces. Heavy falls of rain overwhelm drainage and overfill storage pools. Meteorological observations are infrequent locally, so those made some 70–100 km away were used, along with regridded historical data, reanalysis and CMIP6 projections. These revealed increases in temperature, precipitation and humidity, and provided indications of long-term climate pressures at the site. Changing patterns of future precipitation, particularly heavy rain, may threaten the site. The relative humidity changes are small, though an increasing Scheffer index suggests potentially enhanced wood decay. Changes in soil moisture have the potential to disrupt the foundations. Very hot days may become a problem for the two annual celebrations at the site.
High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (Paphiopedilum concolor × Paphiopedilum hirsutissimum)
Authors: Li Dong-Mei , Zhu Gen-Fa
Published: September 2022

Lady’s slipper orchids (Paphiopedilum spp.) are highly valuable within the flower industry. Recently, both Paphiopedilum concolor and Paphiopedilum hirsutissimum (2n = 2x = 26) have been widely used for hybrid parents, ornamental, and economic purposes. However, high-density genetic maps and leaf traits related to quantitative trait loci (QTLs) in these two Paphiopedilum species have been poorly studied. Herein, an interspecific F1 population of 95 individuals was developed from the cross between P. concolor and P. hirsutissimum with contrasting leaf length (LL), leaf width (LW), leaf thickness (LT), and leaf number (LN). RNA extracted from the F1 population and their parents was subjected to high-throughput RNA sequencing. Approximately 745.59 Gb of clean data were generated, and were assembled into 349,730 transcripts and 185,089 unigenes. In total, 165,196 high-resolution polymorphic single nucleotide polymorphism (SNP) markers were initially identified. Finally, 8410 SNP markers satisfied the requirements and were used to construct a genetic map. The integrated map contained 13 linkage groups (LGs) and spanned 1616.18 cM, with an average distance of 0.19 cM between adjacent markers. QTL analysis in the F1 population identified 12 QTLs linked to four leaf-related traits, including LL, LW, LT, and LN. These QTLs by composition interval mapping, explained 11.86% to 21.58% of the phenotypic variance, and were distributed on eight LGs, but not on LGs 4, 6, 8, 12, and 13. Furthermore, 25 unigenes were identified via BLAST searches between the SNP markers in the QTL regions and our assembled transcriptome, of which 11 unigenes were enriched with 59 gene ontology (GO) terms. The information generated in this study will be useful for candidate genes for further molecular regulation studies on leaf traits, future marker-assisted selection of leaf ornamental improvement breeding, genome assembly, and comparative genome analyses.
Mutation of GmIPK1 Gene Using CRISPR/Cas9 Reduced Phytic Acid Content in Soybean Seeds
Authors: Song Ji Hyeon , Shin Gilok , Kim Hye Jeong , Lee Saet Buyl , Moon Ju Yeon , Jeong Jae Cheol , Choi Hong-Kyu , Kim In Ah , Song Hyeon Jin , Kim Cha Young , Chung Young-Soo
Published: September 2022

Phytic acid (PA) acts as an antinutrient substance in cereal grains, disturbing the bioavailability of micronutrients, such as iron and zinc, in humans, causing malnutrition. GmIPK1 encodes the inositol 1,3,4,5,6-pentakisphosphate 2-kinase enzyme, which converts myo-inopsitol-1,3,4,5,6-pentakisphosphate (IP5) to myo-inositol-1,2,3,4,5,6-hexakisphosphate (IP6) in soybean (Glycine max L.). In this study, for developing soybean with low PA levels, we attempted to edit the GmIPK1 gene using the CRISPR/Cas9 system to introduce mutations into the GmIPK1 gene with guide RNAs in soybean (cv. Kwangankong). The GmIPK1 gene was disrupted using the CRISPR/Cas9 system, with sgRNA-1 and sgRNA-4 targeting the second and third exon, respectively. Several soybean Gmipk1 gene-edited lines were obtained in the T0 generation at editing frequencies of 0.1–84.3%. Sequencing analysis revealed various indel patterns with the deletion of 1–9 nucleotides and insertions of 1 nucleotide in several soybean lines (T0). Finally, we confirmed two sgRNA-4 Gmipk1 gene-edited homozygote soybean T1 plants (line #21-2: 5 bp deletion; line #21-3: 1 bp insertion) by PPT leaf coating assay and PCR analysis. Analysis of soybean Gmipk1 gene-edited lines indicated a reduction in PA content in soybean T2 seeds but did not show any defects in plant growth and seed development.
Disclosing the Biocide Activity of α-Ag2−2xCuxWO4 (0 ≤ x ≤ 0.16) Solid Solutions
Authors: Pereira Paula Fabiana dos Santos , De Foggi Camila Cristina , Gouveia Amanda Fernandes , Pinatti Ivo Mateus , Cabral Luís Antônio , Guillamon Eva , Sorribes Iván , San-Miguel Miguel A. , Vergani Carlos Eduardo , Simões Alexandre Zirpoli , da Silva Edison Z. , Cavalcante Laécio Santos , Llusar Rosa , Longo Elson , Andrés Juan
Published: September 2022

In this work, α-Ag2−2xCuxWO4 (0 ≤ x ≤ 0.16) solid solutions with enhanced antibacterial (against methicillin-resistant Staphylococcus aureus) and antifungal (against Candida albicans) activities are reported. A plethora of techniques (X-ray diffraction with Rietveld refinements, inductively coupled plasma atomic emission spectrometry, micro-Raman spectroscopy, attenuated total reflectance–Fourier transform infrared spectroscopy, field emission scanning electron microscopy, ultraviolet–visible spectroscopy, photoluminescence emissions, and X-ray photoelectron spectroscopy) were employed to characterize the as-synthetized samples and determine the local coordination geometry of Cu2+ cations at the orthorhombic lattice. To find a correlation between morphology and biocide activity, the experimental results were sustained by first-principles calculations at the density functional theory level to decipher the cluster coordinations and electronic properties of the exposed surfaces. Based on the analysis of the under-coordinated Ag and Cu clusters at the (010) and (101) exposed surfaces, we propose a mechanism to explain the biocide activity of these solid solutions.
Latest Advances in Metasurfaces for SERS and SEIRA Sensors as Well as Photocatalysis
Authors: Barbillon Grégory
Published: September 2022

Metasurfaces can enable the confinement of electromagnetic fields on huge surfaces and zones, and they can thus be applied to biochemical sensing by using surface-enhanced Raman scattering (SERS) and surface-enhanced infrared absorption (SEIRA). Indeed, these metasurfaces have been examined for SERS and SEIRA sensing thanks to the presence of a wide density of hotspots and confined optical modes within their structures. Moreover, some metasurfaces allow an accurate enhancement of the excitation and emission processes for the SERS effect by supporting resonances at frequencies of these processes. Finally, the metasurfaces allow the enhancement of the absorption capacity of the solar light and the generation of a great number of catalytic active sites in order to more quickly produce the surface reactions. Here, we outline the latest advances in metasurfaces for SERS and SEIRA sensors as well as photocatalysis.
Renal Transcriptome and Metabolome in Mice with Principal Cell-Specific Ablation of the Tsc1 Gene: Derangements in Pathways Associated with Cell Metabolism, Growth and Acid Secretion
Authors: Zahedi Kamyar , Barone Sharon , Brooks Marybeth , Murray Stewart Tracy , Casero Robert , Soleimani Manoocher
Published: September 2022

Tuberous sclerosis complex (TSC) is caused by mutations in the hamartin (TSC1) or tuberin (TSC2) genes. Using a mouse model of TSC renal cystogenesis that we have previously described, the current studies delineate the metabolic changes in the kidney and their relation to alterations in renal gene expression. To accomplish this, we compared the metabolome and transcriptome of kidneys from 28-day-old wildtype (Wt) and principal cell-specific Tsc1 KO (Tsc1 KO) mice using targeted 1H nuclear magnetic resonance targeted metabolomic and RNA-seq analyses. The significant changes in the kidney metabolome of Tsc1 KO mice included reductions in the level of several amino acids and significant decreases in creatine, NADH, inosine, UDP-galactose, GTP and myo-inositol levels. These derangements may affect energy production and storage, signal transduction and synthetic pathways. The pertinent derangement in the transcriptome of Tsc1 KO mice was associated with increased collecting duct acid secretion, active cell division and the up-regulation of signaling pathways (e.g., MAPK and AKT/PI3K) that suppress the TSC2 GTPase-activating function. The combined renal metabolome and transcriptome alterations observed in these studies correlate with the unregulated growth and predominance of genotypically normal A-intercalated cells in the epithelium of renal cysts in Tsc1 KO mice.
Effect of cx-DHED on Abnormal Glucose Transporter Expression Induced by AD Pathologies in the 5xFAD Mouse Model
Authors: Kim Jinho , Kang ShinWoo , Chang Keun-A
Published: September 2022

Alzheimer’s disease (AD) is a form of dementia associated with abnormal glucose metabolism resulting from amyloid-beta (Aβ) plaques and intracellular neurofibrillary tau protein tangles. In a previous study, we confirmed that carboxy-dehydroevodiamine∙HCl (cx-DHED), a derivative of DHED, was effective at improving cognitive impairment and reducing phosphorylated tau levels and synaptic loss in an AD mouse model. However, the specific mechanism of action of cx-DHED is unclear. In this study, we investigated how the cx-DHED attenuates AD pathologies in the 5xFAD mouse model, focusing particularly on abnormal glucose metabolism. We analyzed behavioral changes and AD pathologies in mice after intraperitoneal injection of cx-DHED for 2 months. As expected, cx-DHED reversed memory impairment and reduced Aβ plaques and astrocyte overexpression in the brains of 5xFAD mice. Interestingly, cx-DHED reversed the abnormal expression of glucose transporters in the brains of 5xFAD mice. In addition, otherwise low O-GlcNac levels increased, and the overactivity of phosphorylated GSK-3β decreased in the brains of cx-DHED-treated 5xFAD mice. Finally, the reduction in synaptic proteins was found to also improve by treatment with cx-DHED. Therefore, we specifically demonstrated the protective effects of cx-DHED against AD pathologies and suggest that cx-DHED may be a potential therapeutic drug for AD.
Gene Expression and Chondrogenic Potential of Cartilage Cells: Osteoarthritis Grade Differences
Authors: Mazor Marija , Lespessailles Eric , Best Thomas M. , Ali Mazen , Toumi Hechmi
Published: September 2022

Recent data suggest that cells isolated from osteoarthritic (OA) cartilage express mesenchymal progenitor cell (MPC) markers that have the capacity to form hyaline-like cartilage tissue. Whether or not these cells are influenced by the severity of OA remains unexplored. Therefore, we analyzed MPC marker expression and chondrogenetic potential of cells from mild, moderate and severe OA tissue. Human osteoarthritic tibial plateaus were obtained from 25 patients undergoing total knee replacement. Each sample was classified as mild, moderate or severe OA according to OARSI scoring. mRNA expression levels of MPC markers—CD105, CD166, Notch 1, Sox9; mature chondrocyte markers—Aggrecan (Acan), Col II A1, hypertrophic chondrocyte and osteoarthritis-related markers—Col I A1, MMP-13 and ALPL were measured at the tissue level (day 0), after 2 weeks of in vitro expansion (day 14) and following chondrogenic in vitro re-differentiation (day 35). Pellet matrix composition after in vitro chondrogenesis of different OA-derived cells was tested for proteoglycans, collagen II and I by safranin O and immunofluorescence staining. Multiple MPC markers were found in OA cartilage resident tissue within a single OA joint with no significant difference between grades except for Notch1, which was higher in severe OA tissues. Expression levels of CD105 and Notch 1 were comparable between OA cartilage-derived cells of different disease grades and bone marrow mesenchymal stem cell (BM-MSC) line (healthy control). However, the MPC marker Sox 9 was conserved after in vitro expansion and significantly higher in OA cartilage-derived cells compared to its levels in the BM-MSC. The in vitro expansion of cartilage-derived cells resulted in enrichment while re–differentiation in reduction of MPC markers for all three analyzed grades. However, only moderate OA-derived cells after the in vitro chondrogenesis resulted in the formation of hyaline cartilage-like tissue. The latter tissue samples were also highly positive for collagen II and proteoglycans with no expression of osteoarthritis-related markers (collagen I, ALPL and MMP13). MPC marker expression did not differ between OA grades at the tissue level. Interestingly after in vitro re-differentiation, only moderate OA-derived cells showed the capacity to form hyaline cartilage-like tissue. These findings may have implications for clinical practice to understand the intrinsic repair capacity of articular cartilage in OA tissues and raises the possibility of these progenitor cells as a candidate for articular cartilage repair.
Restoration of Cathepsin D Level via L-Serine Attenuates PPA-Induced Lysosomal Dysfunction in Neuronal Cells
Authors: Jeon Hyunbum , Kim Yeo Jin , Hwang Su-Kyeong , Seo Jinsoo , Mun Ji Young
Published: September 2022

L-serine is a non-essential amino acid endogenously produced by astrocytes and is abundant in human diets. Beneficial roles of the metabolic products from L-serine in various conditions in the brain including neuronal development have been reported. Through several preclinical studies, L-serine treatment was also shown to offer beneficial therapeutic effects for brain damage such as ischemic stroke, amyotrophic lateral sclerosis, and Parkinson’s disease. Despite evidence for the value of L-serine in the clinic, however, its beneficial effects on the propionic acid (PPA)-induced neuronal toxicity and underlying mechanisms of L-serine-mediated neuroprotection are unknown. In this study, we observed that PPA-induced acidic stress induces abnormal lipid accumulation and functional defects in lysosomes of hippocampal neurons. L-serine treatment was able to rescue the structure and function of lysosomes in PPA-treated hippocampal neuronal cells. We further identified that L-serine suppressed the formation of lipid droplets and abnormal lipid membrane accumulations inside the lysosomes in PPA-treated hippocampal neuronal cells. Taken together, these findings indicate that L-serine can be utilized as a neuroprotective agent for the functionality of lysosomes through restoration of cathepsin D in disease conditions.
Effects of Graphene Oxide and Reduced Graphene Oxide Nanostructures on CD4+ Th2 Lymphocytes
Authors: Feito María José , Cicuéndez Mónica , Casarrubios Laura , Diez-Orejas Rosalía , Fateixa Sara , Silva Daniela , Barroca Nathalie , Marques Paula A. A. P. , Portolés María Teresa
Published: September 2022

The activation of T helper (Th) lymphocytes is necessary for the adaptive immune response as they contribute to the stimulation of B cells (for the secretion of antibodies) and macrophages (for phagocytosis and destruction of pathogens) and are necessary for cytotoxic T-cell activation to kill infected target cells. For these issues, Th lymphocytes must be converted into Th effector cells after their stimulation through their surface receptors TCR/CD3 (by binding to peptide-major histocompatibility complex localized on antigen-presenting cells) and the CD4 co-receptor. After stimulation, Th cells proliferate and differentiate into subpopulations, like Th1, Th2 or Th17, with different functions during the adaptative immune response. Due to the central role of the activation of Th lymphocytes for an accurate adaptative immune response and considering recent preclinical advances in the use of nanomaterials to enhance T-cell therapy, we evaluated in vitro the effects of graphene oxide (GO) and two types of reduced GO (rGO15 and rGO30) nanostructures on the Th2 lymphocyte cell line SR.D10. This cell line offers the possibility of studying their activation threshold by employing soluble antibodies against TCR/CD3 and against CD4, as well as the simultaneous activation of these two receptors. In the present study, the effects of GO, rGO15 and rGO30 on the activation/proliferation rate of these Th2 lymphocytes have been analyzed by studying cell viability, cell cycle phases, intracellular content of reactive oxygen species (ROS) and cytokine secretion. High lymphocyte viability values were obtained after treatment with these nanostructures, as well as increased proliferation in the presence of rGOs. Moreover, rGO15 treatment decreased the intracellular ROS content of Th2 cells in all stimulated conditions. The analysis of these parameters showed that the presence of these GO and rGO nanostructures did not alter the response of Th2 lymphocytes.
NLRP3 Inflammasome/Pyroptosis: A Key Driving Force in Diabetic Cardiomyopathy
Authors: Zhang Lixia , Ai Chenchen , Bai Ming , Niu Jinglei , Zhang Zheng
Published: September 2022

Diabetic cardiomyopathy (DCM), a serious diabetic complication, is a kind of low-grade inflammatory cardiovascular disorder. Due to the high risk of morbidity and mortality, DCM has demanded the attention of medical researchers worldwide. The pathophysiological nature of DCM is intricate, and the genesis and development of which are a consequence of the coaction of many factors. However, the exact pathogenesis mechanism of DCM remains unclear. Pyroptosis is a newly identified programmed cell death (PCD) that is directly related to gasdermin D(GSDMD). It is characterized by pore formation on the cell plasma membrane, the release of inflammatory mediators, and cell lysis. The initiation of pyroptosis is closely correlated with NOD-like receptor 3 (NLRP3) activation, which activates caspase-1 and promotes the cleaving of GSDMD. In addition to adjusting the host’s immune defense, NLRP3 inflammasome/pyroptosis plays a critical role in controlling the systemic inflammatory response. Recent evidence has indicated that NLRP3 inflammasome/pyroptosis has a strong link with DCM. Targeting the activation of NLRP3 inflammasome or pyroptosis may be a hopeful therapeutic strategy for DCM. The focus of this review is to summarize the relevant mechanisms of pyroptosis and the relative contributions in DCM, highlighting the potential therapeutic targets in this field.
Apolipoproteins—New Biomarkers of Overweight and Obesity among Childhood Acute Lymphoblastic Leukemia Survivors?
Authors: Sztolsztener Klaudia , Żywno Hubert , Hodun Katarzyna , Konończuk Katarzyna , Muszyńska-Rosłan Katarzyna , Latoch Eryk
Published: September 2022

Patients suffering from childhood acute lymphoblastic leukemia (ALL) are at risk of late adverse treatment-related effects. The examination of targeted biomarkers could be used to improve the diagnosis and prediction of life-threatening ALL sequelae. The purpose of this cross-sectional study was to search for treatment-related alterations in apolipoprotein (Apo) levels as potential markers of the occurrence of obesity in subjects treated for ALL, and to assess the relationships between weight, gender, anticancer treatment, and Apo concentrations. Fifty-eight ALL survivors were included in the study. The mean time of follow-up after treatment cessation was 5.41 ± 4.29 years. Serum levels of apolipoproteins were measured using a multiplex assay kit. Among ALL survivors, we observed a significant correlation of Apo-C1, Apo-C3, Apo-H, and Apo-J levels, depending on body mass index (BMI). Marked differences were observed in the area under the curve of Apo-A1, Apo-A2, Apo-C1, Apo-D. In our study, patients with a history of childhood ALL developed alterations in their Apo profile. Furthermore, this is the first study revealing that some apolipoproteins may act as valuable biomarkers useful in the prognosis of metabolic imbalance. We believe that this paper, at least partially, will highlight the importance of long-term prognosis of metabolic complications associated with the anticancer chemotherapy used to treat hematological malignancies in children.
Cardiovascular Disease-Associated MicroRNAs as Novel Biomarkers of First-Trimester Screening for Gestational Diabetes Mellitus in the Absence of Other Pregnancy-Related Complications
Authors: Hromadnikova Ilona , Kotlabova Katerina , Krofta Ladislav
Published: September 2022

We assessed the diagnostic potential of cardiovascular disease-associated microRNAs for the early prediction of gestational diabetes mellitus (GDM) in singleton pregnancies of Caucasian descent in the absence of other pregnancy-related complications. Whole peripheral venous blood samples were collected within 10 to 13 weeks of gestation. This retrospective study involved all pregnancies diagnosed with only GDM (n = 121) and 80 normal term pregnancies selected with regard to equality of sample storage time. Gene expression of 29 microRNAs was assessed using real-time RT-PCR. Upregulation of 11 microRNAs (miR-1-3p, miR-20a-5p, miR-20b-5p, miR-23a-3p, miR-100-5p, miR-125b-5p, miR-126-3p, miR-181a-5p, miR-195-5p, miR-499a-5p, and miR-574-3p) was observed in pregnancies destinated to develop GDM. Combined screening of all 11 dysregulated microRNAs showed the highest accuracy for the early identification of pregnancies destinated to develop GDM. This screening identified 47.93% of GDM pregnancies at a 10.0% false positive rate (FPR). The predictive model for GDM based on aberrant microRNA expression profile was further improved via the implementation of clinical characteristics (maternal age and BMI at early stages of gestation and an infertility treatment by assisted reproductive technology). Following this, 69.17% of GDM pregnancies were identified at a 10.0% FPR. The effective prediction model specifically for severe GDM requiring administration of therapy involved using a combination of these three clinical characteristics and three microRNA biomarkers (miR-20a-5p, miR-20b-5p, and miR-195-5p). This model identified 78.95% of cases at a 10.0% FPR. The effective prediction model for GDM managed by diet only required the involvement of these three clinical characteristics and eight microRNA biomarkers (miR-1-3p, miR-20a-5p, miR-20b-5p, miR-100-5p, miR-125b-5p, miR-195-5p, miR-499a-5p, and miR-574-3p). With this, the model identified 50.50% of GDM pregnancies managed by diet only at a 10.0% FPR. When other clinical variables such as history of miscarriage, the presence of trombophilic gene mutations, positive first-trimester screening for preeclampsia and/or fetal growth restriction by the Fetal Medicine Foundation algorithm, and family history of diabetes mellitus in first-degree relatives were included in the GDM prediction model, the predictive power was further increased at a 10.0% FPR (72.50% GDM in total, 89.47% GDM requiring therapy, and 56.44% GDM managed by diet only). Cardiovascular disease-associated microRNAs represent promising early biomarkers to be implemented into routine first-trimester screening programs with a very good predictive potential for GDM.
Salivary Exosomal miRNA-1307-5p Predicts Disease Aggressiveness and Poor Prognosis in Oral Squamous Cell Carcinoma Patients
Authors: Patel Aditi , Patel Shanaya , Patel Parina , Mandlik Dushyant , Patel Kaustubh , Tanavde Vivek
Published: September 2022

Background: Salivary exosomal miRNAs as biomarkers facilitate repeated sampling, real-time disease monitoring and assessment of therapeutic response. This study identifies a single salivary exosomal miRNA prognosticator that will aid in improved patient outcome using a liquid biopsy approach. Method: Small RNA and transcriptome sequencing profiles of tumour tissues (n = 12) and salivary exosomes (n = 8) from oral cancer patients were compared to their non-cancerous counterparts. We validated these results using The Cancer Genome Atlas database and performing Real-time PCR on a large patient cohort (n = 19 tissue samples; n = 12 salivary exosomes). Potential target genes and the miRNA–mRNA networks and enriched biological pathways regulated by this microRNA were identified using computational tools. Results: Salivary exosomes (size: 30–50 nm) demonstrated a strong expression of CD47 and detectable expression of tetraspanins CD63, CD81 and CD9 by flow cytometry. miR-1307-5p was exclusively overexpressed in tissues and salivary exosomes of oral cancer patients compared to their non-cancerous counterparts. Enhanced expression of miR-1307-5p clinically correlated with poor patient survival, disease progression, aggressiveness and chemo-resistance. Transcriptome analysis suggested that miRNA-1307-5p could promote oral cancer progression by suppressing THOP1, EHF, RNF4, GET4 and RNF114. Conclusions: Salivary exosomal miRNA-1307-5p is a potential prognosticator for predicting poor survival and poor patient outcome in oral cancers.
Replication of Human Norovirus in Mice after Antibiotic-Mediated Intestinal Bacteria Depletion
Authors: Santiso-Bellón Cristina , Gozalbo-Rovira Roberto , Buesa Javier , Rubio-del-Campo Antonio , Peña-Gil Nazaret , Navarro-Lleó Noemi , Cárcamo-Calvo Roberto , Yebra María J. , Monedero Vicente , Rodríguez-Díaz Jesús
Published: September 2022

Human noroviruses (HuNoVs) are the main cause of acute gastroenteritis causing more than 50,000 deaths per year. Recent evidence shows that the gut microbiota plays a key role in enteric virus infectivity. In this context, we tested whether microbiota depletion or microbiota replacement with that of human individuals susceptible to HuNoVs infection could favor viral replication in mice. Four groups of mice (n = 5) were used, including a control group and three groups that were treated with antibiotics to eliminate the autochthonous intestinal microbiota. Two of the antibiotic-treated groups received fecal microbiota transplantation from a pool of feces from infants (age 1–3 months) or an auto-transplantation with mouse feces that obtained prior antibiotic treatment. The inoculation of the different mouse groups with a HuNoVs strain (GII.4 Sydney [P16] genotype) showed that the virus replicated more efficiently in animals only treated with antibiotics but not subject to microbiota transplantation. Viral replication in animals receiving fecal microbiota from newborn infants was intermediate, whereas virus excretion in feces from auto-transplanted mice was as low as in the control mice. The analysis of the fecal microbiota by 16S rDNA NGS showed deep variations in the composition in the different mice groups. Furthermore, differences were observed in the gene expression of relevant immunological mediators, such as IL4, CXCL15, IL13, TNFα and TLR2, at the small intestine. Our results suggest that microbiota depletion eliminates bacteria that restrict HuNoVs infectivity and that the mechanism(s) could involve immune mediators.
Metabolic and Proteomic Profiles Associated with Immune Responses Induced by Different Inactivated SARS-CoV-2 Vaccine Candidates
Authors: Yu Shouzhi , He Yao , Ji Wenheng , Yang Rong , Zhao Yuxiu , Li Yan , Liu Yingwei , Ding Ling , Ma Meng , Wang Hui , Yang Xiaoming
Published: September 2022

Since the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in late 2019, the virus has been mutating continuously, resulting in the continuous emergence of variants and creating challenges for epidemic prevention and control. Here, we immunized mice with different vaccine candidates, revealing the immune, protein, and metabolomic changes that take place in vaccines composed of different variants. We found that the prototype strain and Delta- and Omicron-variant inactivated vaccine candidates could all induce a high level of neutralizing antibodies and cellular immunity responses in mice. Next, we found that the metabolic and protein profiles were changed, showing a positive association with immune responses, and the level of the change was distinct in different inactivated vaccines, indicating that amino acid variations could affect metabolomics and proteomics. Our findings reveal differences between vaccines at the metabolomic and proteomic levels. These insights provide a novel direction for the immune evaluation of vaccines and could be used to guide novel strategies for vaccine design.