bluuebunny/crossref_metadata_2025 huggingface dataset:
Variability in the Hydrological Processes of Six Typical Woodlands Based on Stable Isotopes in Subtropical Regions in Central China
Authors: Zhu Fangfang , Li Yuchen , Cheng Jinhua
Published: June 2023

Changes in woodland characteristics induced by plants and soil greatly affect soil hydrological processes. Stable isotope technology and indoor soil moisture characteristic experiments were conducted at three rainfall levels (3.6, 23.5, and 49.8 mm) to investigate the hydrological processes under six woodlands (two pure forests and four mixed forests). The main influencing factors contributing to these changes were identified in a low mountain and hilly region in central China. The soil waterline equation in this area was soil water δD = 5.626 δ18O − 16.791 (R2 = 0.798). The slope and intercept in the soil waterline equation were smaller than the atmospheric waterline equation. From a temporal perspective, the soil moisture content varied in the same trend under different rainfall events, with the maximum and minimum values on the first day after rainfall and the day before rainfall, respectively. However, an overall trend that first increased and then decreased was observed. From a spatial perspective, the soil moisture content increased with soil depth, and the increase rate was in the order of 0–20 cm and 20–40 cm in different soil layers. The soil moisture content in mixed conifer broadleaved woodlands was high. The soil water δD and δ18O in mixed conifer broadleaved woodlands and underground soil were relatively depleted. The effects of soil water-holding capacity, particle size composition, slope, canopy closure, and other factors on soil hydraulic parameters were comprehensively analyzed. The results showed that the extremely coarse sand (1–2 mm) particle content was the main parameter affecting soil-saturated hydraulic conductivity Ks, whereas the slope was the main factor affecting soil water δD and δ18O. In needle-leaved forests, the soil water infiltration form was a rainwater and soil water mixture downward diffusion, whereas the rainwater replaced the original soil water in the needle and mixed conifer broadleaved forests.
VOC Emission from Lightweight Wood Fiber Insulation Board
Authors: Fuczek Dorota , Czajka Magdalena , Szuta Jarosław , Szutkowski Krystian , Kwaśniewska-Sip Patrycja
Published: June 2023

The aim of the presented research work was to determine and analyze emissions of volatile organic compounds (VOCs) from experimental lightweight wood fiber insulation board produced in dry technology. Until now, there have been no rigid insulation materials made of wood fibers produced in such low density and made in dry technology. Among the typical parameters such as thermal conductivity and the mechanical performance of the lightweight board, attention was also paid to their influence on indoor air quality. Therefore, an attempt was made to determine the kind of substances emitting from wood fiber insulation boards produced at defined production parameters as well as the dynamics of emission reduction over time. Additionally, the influence of fire retardants used for protection against lightweight wood fiberboard fires on the emission of VOCs was analyzed. Tests on VOC emissions were carried out using the chamber method according to the applicable ISO 16000 standards. The main components emitting from lightweight insulation fiberboards were acetic acid and aldehydes such as pentanal, hexanal, heptanal, octanal, nonanal, decanal, furfural, and benzaldehyde. The percentage of acetic acid in total volatile organic compounds (TVOCs) was within the limits of 17% to 65%. From the aldehydes group, the most concerning substance was furfural due to a very strict limit value. In the presented research, depending on the variant, the emission of furfural was from 0 up to 10 µg/m3 after 28 days of measurement. Other substances such as terpenes or aromatic hydrocarbons were at a very low level. The reduction in VOCs over a period of 28 days was significant in most cases from 22% up to 61%. The tests carried out also showed a substantial impact of fire retardant, used in the production of lightweight insulation fiberboard, on the emission of VOCs from fiberboards, and thus on their quality.
An Improved Sap Flow Prediction Model Based on CNN-GRU-BiLSTM and Factor Analysis of Historical Environmental Variables
Authors: Li Yane , Guo Lijun , Wang Jiyang , Wang Yiwei , Xu Dayu , Wen Jun
Published: June 2023

Sap flow is widely used to estimate the transpiration and water consumption of canopies and to manage water resources. In this paper, an improved time series prediction model was proposed by integrating three basic networks—CNN, GRU and BiLSTM—to assess sap flow with historical environment variables. A dataset with 17,569 records of each, including 9 environment variables and 1 sap flow, was applied from a public database of SAPFLUXNET. After normalization, the environment variables were analyzed and composed with the factor analysis method. After the CNN-GRU-BiLSTM structure was designed, N records of three main factors were computed from environment variables, which were measured at N previous moments, and the sap flow was measured at the current moment, and they were applied for each training, validation, and testing cycle. To improve and compare the CNN-GRU-BiLSTM-based model, nine other models, using the methods of multiple linear regression, support vector regression, random forest, LSTM, GRU, BiLSTM, CNN-GRU, CNN-BiLSTM, and CNN-GRU-LSTM, were constructed in this study, respectively. Results show that the performance of the CNN-GRU-BiLSTM-based model has more accuracy than the other nine models we built in this paper, with the mean absolute error, mean squared error, mean absolute percentage error, and coefficient of determination (R2) being 0.0410, 0.0029, 0.2708 and 0.9329, respectively. Furthermore, for a comparison of the descending dimension method of factor analysis, principal component analysis (PCA) and singular value decomposition (SVD) methods were applied and compared, respectively. Results show that the performance of the factor analysis-based model is better than the PCA- or SVD-based model, with the R2 results of the factor analysis-based model being higher than the PCA- and SVD-based models by 5.06% and 10.63%, respectively. This study indicates that the CNN-GRU-BiLSTM-based sap flow prediction model established with a factor analysis of historical environmental variables has optimistic applications for analyzing the transpiration of trees and evaluating water consumption.
A Case Study on the Effects of Weather Conditions on Forest Fire Propagation Parameters in the Malekroud Forest in Guilan, Iran
Authors: Mohammadian Bishe Esmaeil , Norouzi Mohammad , Afshin Hossein , Farhanieh Bijan
Published: June 2023

The present study investigates the effect of climatic parameters, such as air relative humidity and wind speed, on fire spread propagation indexes in the Malekroud Forest, Iran using the FARSITE simulator based on Rothermel’s original fire spread equation. Standard fuel models are used to calibrate the vegetation cover. Sorensen (SC) and kappa (κ) coefficients, as well as the Overestimation Index (OI), are used to estimate the simulation’s accuracy. The results confirm that using both ambient condition data and appropriate fuel models is crucial to reaching reasonable results in fire propagation simulations. The values of the Rate of Fire Spread (ROS), Flame Length (FML), and Fire Line Intensity (FLI) are reported for each particular scenario. The simulation results show that the Sorensen and Kappa coefficient for situations most similar to the real fire reached 0.82 and 0.80, respectively. The investigated fire’s severity is categorized as low-condition fire behavior. The simulation shows that fire propagation falls harshly in the case of air relative humidity by more than 72%, and we will not witness natural fire propagation on a large scale.
Mediterranean Diet, Sustainability, and Tourism—A Study of the Market’s Demand and Knowledge
Authors: Ingrassia Marzia , Altamore Luca , Columba Pietro , Raffermati Sara , Lo Grasso Giuseppe , Bacarella Simona , Chironi Stefania
Published: June 2023

Globalization intensified competitiveness among agribusinesses worldwide in recent years. The European Commission focused on enhancing sustainable agriculture and food products’ territorial uniqueness for competing in the international market. The Mediterranean diet (MD) is a model of feeding and lifestyle belonging to the ancient Mediterranean culture, which also embodies a sustainable food system. Therefore, in 2010, UNESCO recognized the MD as Intangible Cultural Heritage of Humanity, and Sicily (southern Italy) is its official physical site. Despite its worldwide fame, the notion of the MD runs the risk of being mystified because it is described most often as something that does not correspond to what it is holistically. The aim of this study is to know the market demand of the MD by Italian people and foreign tourists in Sicily and the level of knowledge of the MD by users, both consumers and experts. A survey at top Sicilian traditional restaurants with owners/chefs and their clients was carried out. The study provided an in-depth understanding of the current lack of knowledge about the holistic meaning of the MD. The study highlights the desirability of integrated science–policy actions (also for communication) and proposes a vertical communication system to revive and direct the MD demand toward its holistic model.
Influence of a Commercial Synbiotic Administered In Ovo and In-Water on Broiler Chicken Performance and Meat Quality
Authors: Tavaniello Siria , De Marzo Davide , Bednarczyk Marek , Palazzo Marisa , Zejnelhoxha Sanije , Wu Mengjun , Peng Meng , Stadnicka Katarzyna , Maiorano Giuseppe
Published: June 2023

The present study aimed to test the synbiotic PoultryStar® solUS delivered in ovo to evaluate its effect on hatchability, productive performance and meat quality, compared to its post-hatch administration in water. On the twelfth day of embryonic incubation, 1200 fertile eggs were divided into synbiotic groups injected with 2 mg/embryo (T1) and 3 mg/embryo (T2), a saline group injected with physiological saline and an uninjected control group (C). After hatching, 120 male chicks/group were reared and chicks from the saline group were supplemented with the synbiotic via drinking water (T3). Hatchability was low in both T1 and T2 groups. Growth performance was not affected by the treatments. However, in the second rearing phase (15–36 days), birds from the C and T3 groups were heavier than T1 birds, due to a higher feed intake and daily weight gain. Neither route of synbiotic administration influenced final body weight (at 56 days), weight and yield of the carcass or commercial cuts. Physico-chemical properties, total lipid, cholesterol and fatty acid composition of breast muscle were not affected by the treatments. Considering its exploratory nature, this study has raised many questions that need further investigation, such as the bioactive combination and the effect on embryonic development.
Effects of Inner-Row Ground Management on the Volatomics of ‘Cabernet Sauvignon’ Grapes and Wines in the Region of the Eastern Foothills of the Ningxia Helan Mountains in Northwest China
Authors: He Fei , Tian Meng-Bo , Duan Wei-Peng , Yang Wei-Ming , Mao Xue , Wang Jun , Duan Chang-Qing
Published: June 2023

This two-consecutive-year study aimed to evaluate the effects of ground management methods on the volatomics of ‘Cabernet Sauvignon’ grapes and wines in Northwest China, in which inner-row crop covering with purslane (GRASS) and mulching with black plastic film (FILM) treatments were carried out, respectively. Compared with clean tillage (CK), the GRASS and FILM treatments changed the microclimates of grapevine fruit zones and rhizospheres, which delayed the ripening of grape berries and affected the accumulation of aroma substances in the mature grapes effectively. GRASS increased the concentration of terpenes and C13-norisoprenoids in berries and gave more floral, fruity, and caramel fragrances to wines, while FILM had the opposite effect of significantly increasing the synthesis of C6/C9 compounds and brought more green leaf flavors, showing that inner-row purslane covering is a potential and stable viticultural practice to improve the wine quality in this booming wine region.
Shotgun Metagenomic Sequencing Revealed the Prebiotic Potential of a Fruit Juice Drink with Fermentable Fibres in Healthy Humans
Authors: Bester Adri , O’Brien Marcus , Cotter Paul , Dam Sarita , Civai Claudia
Published: June 2023

Fibre-based dietary interventions are at the forefront of gut microbiome modulation research, with a wealth of 16S rRNA information to demonstrate the prebiotic effects of isolated fibres. However, there is a distinct lack of data relating to the effect of a combination of soluble and insoluble fibres in a convenient-to-consume fruit juice food matrix on gut microbiota structure, diversity, and function. Here, we aimed to determine the impact of the MOJU Prebiotic Shot, an apple, lemon, ginger, and raspberry fruit juice drink blend containing chicory inulin, baobab, golden kiwi, and green banana powders, on gut microbiota structure and function. Healthy adults (n = 20) were included in a randomised, double-blind, placebo-controlled, cross-over study, receiving 60 mL MOJU Prebiotic Shot or placebo (without the fibre mix) for 3 weeks with a 3-week washout period between interventions. Shotgun metagenomics revealed significant between-group differences in alpha and beta diversity. In addition, the relative abundance of the phyla Actinobacteria and Desulfobacteria was significantly increased as a result of the prebiotic intervention. Nine species were observed to be differentially abundant (uncorrected p-value of <0.05) as a result of the prebiotic treatment. Of these, Bifidobacterium adolescentis and CAG-81 sp900066785 (Lachnospiraceae) were present at increased abundance relative to baseline. Additionally, KEGG analysis showed an increased abundance in pathways associated with arginine biosynthesis and phenylacetate degradation during the prebiotic treatment. Our results show the effects of the daily consumption of 60 mL MOJU Prebiotic Shot for 3 weeks and provide insight into the functional potential of B. adolescentis.
Genetic Basis of Inflammatory Demyelinating Diseases of the Central Nervous System: Multiple Sclerosis and Neuromyelitis Optica Spectrum
Authors: Ortiz Genaro Gabriel , Torres-Mendoza Blanca M. G. , Ramírez-Jirano Javier , Marquez-Pedroza Jazmin , Hernández-Cruz José J. , Mireles-Ramirez Mario A. , Torres-Sánchez Erandis D.
Published: June 2023

Demyelinating diseases alter myelin or the coating surrounding most nerve fibers in the central and peripheral nervous systems. The grouping of human central nervous system demyelinating disorders today includes multiple sclerosis (MS) and neuromyelitis optica spectrum disorders (NMOSD) as distinct disease categories. Each disease is caused by a complex combination of genetic and environmental variables, many involving an autoimmune response. Even though these conditions are fundamentally similar, research into genetic factors, their unique clinical manifestations, and lesion pathology has helped with differential diagnosis and disease pathogenesis knowledge. This review aims to synthesize the genetic approaches that explain the differential susceptibility between these diseases, explore the overlapping clinical features, and pathological findings, discuss existing and emerging hypotheses on the etiology of demyelination, and assess recent pathogenicity studies and their implications for human demyelination. This review presents critical information from previous studies on the disease, which asks several questions to understand the gaps in research in this field.
One Pandemic, Two Solutions: Comparing the U.S.-China Response and Health Priorities to COVID-19 from the Perspective of “Two Types of Control”
Authors: Lyu Shupeng , Qian Chen , McIntyre Aaron , Lee Ching-Hung
Published: June 2023

After three years of global rampage, the COVID-19 epidemic, the most serious infectious disease to occur worldwide since the 1918 influenza pandemic, is nearing its end. From the global experience, medical control and social control are the two main dimensions in the prevention and control of COVID-19. From the perspective of “two types of control”, namely medical control and social control, this paper finds that the political system, economic structure, and cultural values of the United States greatly limit the government’s ability to impose social control, forcing it to adopt medical control to fight the virus in a single dimension. In contrast, China’s political system, economic structure, and cultural values allow its government to adopt stringent, extensive, and frequent social control, as well as medical control to fight the virus. This approach departs from the traditional pathway of fighting the epidemic, i.e., “infection–treatment–immunization”, thereby outpacing the evolution of the virus and controlling its spread more rapidly. This finding helps explain why the Chinese government adopted a strict “zeroing” and “dynamic zeroing” policy during the first three years, at the cost of enormous economic, social, and even political legitimacy. It was not until late 2022, when the Omicron variant with the waning virulence became prevalent, that China chose to “coexist” with the virus, thus avoiding a massive epidemic-related death. While the United States adopted a pulsed-style strategy at the beginning of the epidemic, i.e., “relaxation–suppression–relaxation–suppression”, and began to “coexist” with the virus in just one year, resulting in a large number of excess deaths associated with the epidemic. The study contributes to explaining the difference in the interplay between public health priorities and COVID-19 response strategies in China and the United States, based on the specific public health context and the perspective of “medical control” and “social control”.
Toward a Holistic Understanding of Factors That Support or Inhibit Graduate Student Success
Authors: Collier Karen M. , Blanchard Margaret R.
Published: June 2023

The goal of this research was to gain a holistic understanding of factors that support or inhibit graduate student success, with a particular interest in the experiences of underrepresented minorities, females in STEM, and first-generation college students. The Graduate Student Success Survey (GSSS) was developed and validated with 537 M.S. and Ph.D. students at a research-intensive university in the southeastern United States. Guided by Maslow’s Hierarchy of Needs and informed by salient factors described in the literature and published surveys, items were developed to measure students’ perceptions of imposter syndrome, microaggressions, microaffirmations, mentoring, sense of belonging, financial support, and mentor relationships. This manuscript details the development of these items and validation of the GSSS with full- and part-time thesis-based graduate students across twelve colleges, six of which were STEM-focused. Validity and reliability were tested with exploratory and confirmatory factor analysis, resulting in a survey with seven subscales. Findings revealed significant differences in these graduate students’ experiences based on their gender identity, racial and ethnic group, citizenship status, and program area. The findings of this study report the experiences of graduate students at one institution. However, the validated survey and the recommendations resulting from the findings could be used to scaffold student success and provide insight for faculty and administrators on how to better support students.
Dysfunctional Schemas from Preadolescence as One Major Avenue by Which Meaning Has Impact on Mental Health
Authors: André Nathalie , Baumeister Roy F.
Published: June 2023

A main way by which meaning influences mental health is by the formation of interpersonal schemas that specify what to expect from others and how to treat them. Particularly during preadolescence (a developmental phase focused on interpersonal skills), young people living in a stressful or hurtful environment can form atypical schemas that can help them survive but that produce serious problems when later applied to newly forming adult relationships. We provide three case studies illustrating this process. A boy learned to cope by withdrawing from social interaction and excelling in schoolwork. A girl learned to cope by denying her own needs and sacrificing herself for the welfare of others. Another girl coped by pervasive distrust of others and by becoming assertively independent. These children learned well enough to adapt to these dysfunctional relationships so as to suffer as little as possible, and they even developed some personal skills and resources. However, the rigid schemas had a destructive impact on their adult relationships. Proposals for interventions to change meaning and behaviors are discussed.
Cross-Kingdom Interaction of miRNAs and Gut Microbiota with Non-Invasive Diagnostic and Therapeutic Implications in Colorectal Cancer
Authors: Pös Ondrej , Styk Jakub , Buglyó Gergely , Zeman Michal , Lukyova Lydia , Bernatova Kamila , Hrckova Turnova Evelina , Rendek Tomas , Csók Ádám , Repiska Vanda , Nagy Bálint , Szemes Tomas
Published: June 2023

Colorectal cancer (CRC) has one of the highest incidences among all types of malignant diseases, affecting millions of people worldwide. It shows slow progression, making it preventable. However, this is not the case due to shortcomings in its diagnostic and management procedure and a lack of effective non-invasive biomarkers for screening. Here, we discuss CRC-associated microRNAs (miRNAs) and gut microbial species with potential as CRC diagnostic and therapy biomarkers. We provide rich evidence of cross-kingdom miRNA-mediated interactions between the host and gut microbiome. miRNAs have emerged with the ability to shape the composition and dynamics of gut microbiota. Intestinal microbes can uptake miRNAs, which in turn influence microbial growth and provide the ability to regulate the abundance of various microbial species. In the context of CRC, targeting miRNAs could aid in manipulating the balance of the microbiota. Our findings suggest the need for correlation analysis between the composition of the gut microbiome and the miRNA expression profile.
Multisensory Stimulation Reverses Memory Impairment in Adrβ3KO Male Mice
Authors: Ravache Thaís T. , Batistuzzo Alice , Nunes Gabriela G. , Gomez Thiago G. B. , Lorena Fernanda B. , Do Nascimento Bruna P. P. , Bernardi Maria Martha , Lima Eduarda R. R. , Martins Daniel O. , Campos Ana Carolina P. , Pagano Rosana L. , Ribeiro Miriam O.
Published: June 2023

Norepinephrine plays an important role in modulating memory through its beta-adrenergic receptors (Adrβ: β1, β2 and β3). Here, we hypothesized that multisensory stimulation would reverse memory impairment caused by the inactivation of Adrβ3 (Adrβ3KO) with consequent inhibition of sustained glial-mediated inflammation. To test this, 21- and 86-day-old Adrβ3KO mice were exposed to an 8-week multisensory stimulation (MS) protocol that comprised gustatory and olfactory stimuli of positive and negative valence; intellectual challenges to reach food; the use of hidden objects; and the presentation of food in ways that prompted foraging, which was followed by analysis of GFAP, Iba-1 and EAAT2 protein expression in the hippocampus (HC) and amygdala (AMY). The MS protocol reduced GFAP and Iba-1 expression in the HC of young mice but not in older mice. While this protocol restored memory impairment when applied to Adrβ3KO animals immediately after weaning, it had no effect when applied to adult animals. In fact, we observed that aging worsened the memory of Adrβ3KO mice. In the AMY of Adrβ3KO older mice, we observed an increase in GFAP and EAAT2 expression when compared to wild-type (WT) mice that MS was unable to reduce. These results suggest that a richer and more diverse environment helps to correct memory impairment when applied immediately after weaning in Adrβ3KO animals and indicates that the control of neuroinflammation mediates this response.
Atheroma-Relevant 7-Oxysterols Differentially Upregulate Cd14 Expression
Authors: Kim Bo-Young , Son Yonghae , Kim Byoung Joon , Chung Sung Woon , Lee Dongjun , Eo Seong-Kug , Kim Koanhoi
Published: June 2023

The expression of CD14 in monocytic cells is elevated in atherosclerotic lesions where 7-oxyterols are abundant. However, it remains unknown whether atheroma-relevant 7-oxysterols are involved in receptor expression. Therefore, we investigated the effects of 7α-hydroxycholesterol (7αOHChol), 7β-hydroxycholesterol (7βOHChol), and 7-ketocholesterol (7K) on CD14 levels in THP-1 cells. The three 7-oxysterols increased CD14 transcript levels at a distinct time point, elevated cellular CD14 protein levels, and promoted the release of soluble CD (sCD14) from THP-1 cells. Our data revealed that CD14 expression was most strongly induced after treatment with 7αOHChol. Moreover, 7αOHChol alone upregulated membrane-bound CD14 levels and enhanced responses to lipopolysaccharides, as determined by CCL2 production and monocytic cell migration. The 7-oxysterols also increased the gelatinolytic activity of MMP-9, and a cell-permeable, reversible MMP-9 inhibitor, MMP-9 inhibitor I, significantly impaired sCD14 release. These results indicate that 7-oxysterols differentially induce CD14 expression in vascular cells and contribute to the monocytic cell expression of CD14 via overlapping, but distinct, mechanisms.
Spermidine Attenuates High Glucose-Induced Oxidative Damage in Retinal Pigment Epithelial Cells by Inhibiting Production of ROS and NF-κB/NLRP3 Inflammasome Pathway
Authors: Bang EunJin , Park Cheol , Hwangbo Hyun , Shim Jung-Hyun , Leem Sun-Hee , Hyun Jin Won , Kim Gi-Young , Choi Yung Hyun
Published: June 2023

Diabetic retinopathy (DR) is the leading cause of vision loss and a critical complication of diabetes with a very complex etiology. The build-up of reactive oxygen species (ROS) due to hyperglycemia is recognized as a primary risk factor for DR. Although spermidine, a naturally occurring polyamine, has been reported to have antioxidant effects, its effectiveness in DR has not yet been examined. Therefore, in this study, we investigated whether spermidine could inhibit high glucose (HG)-promoted oxidative stress in human retinal pigment epithelial (RPE) cells. The results demonstrated that spermidine notably attenuated cytotoxicity and apoptosis in HG-treated RPE ARPE-19 cells, which was related to the inhibition of mitochondrial ROS production. Under HG conditions, interleukin (IL)-1β and IL-18’s release levels were markedly increased, coupled with nuclear factor kappa B (NF-κB) signaling activation. However, spermidine counteracted the HG-induced effects. Moreover, the expression of nucleotide-binding oligomerization domain-like receptor (NLR) protein 3 (NLRP3) inflammasome multiprotein complex molecules, including TXNIP, NLRP3, ASC, and caspase-1, increased in hyperglycemic ARPE-19 cells, but spermidine reversed these molecular changes. Collectively, our findings demonstrate that spermidine can protect RPE cells from HG-caused injury by reducing ROS and NF-κB/NLRP3 inflammasome pathway activation, indicating that spermidine could be a potential therapeutic compound for DR treatment.
Recent Advances of 68Ga-Labeled PET Radiotracers with Nitroimidazole in the Diagnosis of Hypoxia Tumors
Authors: Nguyen Anh Thu , Kim Hee-Kwon
Published: June 2023

Positron emission tomography (PET) is a noninvasive molecular imaging method extensively applied in the detection and treatment of various diseases. Hypoxia is a common phenomenon found in most solid tumors. Nitroimidazole is a group of bioreducible pharmacophores that selectively accumulate in hypoxic regions of the body. Over the past few decades, many scientists have reported the use of radiopharmaceuticals containing nitroimidazole for the detection of hypoxic tumors. Gallium-68, a positron-emitting radioisotope, has a favorable half-life time of 68 min and can be conveniently produced by 68Ge/68Ga generators. Recently, there has been significant progress in the preparation of novel 68Ga-labeled complexes bearing nitroimidazole moieties for the diagnosis of hypoxia. This review provides a comprehensive overview of the current status of developing 68Ga-labeled radiopharmaceuticals with nitroimidazole moieties, their pharmacokinetics, and in vitro and in vivo studies, as well as PET imaging studies for hypoxic tumors.
The Pro-Oncogenic Sphingolipid-Metabolizing Enzyme β-Galactosylceramidase Modulates the Proteomic Landscape in BRAF(V600E)-Mutated Human Melanoma Cells
Authors: Capoferri Davide , Chiodelli Paola , Corli Marzia , Belleri Mirella , Scalvini Elisa , Mignani Luca , Guerra Jessica , Grillo Elisabetta , De Giorgis Veronica , Manfredi Marcello , Presta Marco
Published: June 2023

β-Galactosylceramidase (GALC) is a lysosomal enzyme involved in sphingolipid metabolism by removing β-galactosyl moieties from β-galactosylceramide and β-galactosylsphingosine. Previous observations have shown that GALC may exert pro-oncogenic functions in melanoma and Galc silencing, leading to decreased oncogenic activity in murine B16 melanoma cells. The tumor-driving BRAF(V600E) mutation is present in approximately 50% of human melanomas and represents a major therapeutic target. However, such mutation is missing in melanoma B16 cells. Thus, to assess the impact of GALC in human melanoma in a more relevant BRAF-mutated background, we investigated the effect of GALC overexpression on the proteomic landscape of A2058 and A375 human melanoma cells harboring the BRAF(V600E) mutation. The results obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) demonstrate that significant differences exist in the protein landscape expressed under identical cell culture conditions by A2058 and A375 human melanoma cells, both harboring the same BRAF(V600E)-activating mutation. GALC overexpression resulted in a stronger impact on the proteomic profile of A375 cells when compared to A2058 cells (261 upregulated and 184 downregulated proteins versus 36 and 14 proteins for the two cell types, respectively). Among them, 25 proteins appeared to be upregulated in both A2058-upGALC and A375-upGALC cells, whereas two proteins were significantly downregulated in both GALC-overexpressing cell types. These proteins appear to be involved in melanoma biology, tumor invasion and metastatic dissemination, tumor immune escape, mitochondrial antioxidant activity, endoplasmic reticulum stress responses, autophagy, and/or apoptosis. Notably, analysis of the expression of the corresponding genes in human skin cutaneous melanoma samples (TCGA, Firehose Legacy) using the cBioPortal for Cancer Genomics platform demonstrated a positive correlation between GALC expression and the expression levels of 14 out of the 27 genes investigated, thus supporting the proteomic findings. Overall, these data indicate for the first time that the expression of the lysosomal sphingolipid-metabolizing enzyme GALC may exert a pro-oncogenic impact on the proteomic landscape in BRAF-mutated human melanoma.
Synonymous Variants of Uncertain Silence
Authors: Giacoletto Christopher J. , Rotter Jerome I. , Grody Wayne W. , Schiller Martin R.
Published: June 2023

Synonymous variants, traditionally regarded as silent mutations due to their lack of impact on protein sequence, structure and function, have been the subject of increasing scrutiny. This commentary explores the emerging evidence challenging the notion of synonymous variants as functionally inert. Analysis of the activity of 70 synonymous variants in the HIV Tat transcription factor revealed that 50% of the variants exhibited significant deviations from wild-type activity. Our analysis supports previous work and raises important questions about the broader impact of non-silent synonymous variants in human genes. Considering the potential functional implications, the authors propose classifying such variants as “synonymous variants of uncertain silence” (sVUS), highlighting the need for cautious interpretation and further investigations in clinical and genetic testing settings.
Inhibition of Pertussis Toxin by Human α-Defensins-1 and -5: Differential Mechanisms of Action
Authors: Kling Carolin , Sommer Anja , Almeida-Hernandez Yasser , Rodríguez Armando , Perez-Erviti Julio A. , Bhadane Rajendra , Ständker Ludger , Wiese Sebastian , Barth Holger , Pupo-Meriño Mario , Pulliainen Arto T. , Sánchez-García Elsa , Ernst Katharina
Published: June 2023

Whooping cough is a severe childhood disease, caused by the bacterium Bordetella pertussis, which releases pertussis toxin (PT) as a major virulence factor. Previously, we identified the human antimicrobial peptides α-defensin-1 and -5 as inhibitors of PT and demonstrated their capacity to inhibit the activity of the PT enzyme subunit PTS1. Here, the underlying mechanism of toxin inhibition was investigated in more detail, which is essential for developing the therapeutic potential of these peptides. Flow cytometry and immunocytochemistry revealed that α-defensin-5 strongly reduced PT binding to, and uptake into cells, whereas α-defensin-1 caused only a mild reduction. Conversely, α-defensin-1, but not α-defensin-5 was taken up into different cell lines and interacted with PTS1 inside cells, based on proximity ligation assay. In-silico modeling revealed specific interaction interfaces for α-defensin-1 with PTS1 and vice versa, unlike α-defensin-5. Dot blot experiments showed that α-defensin-1 binds to PTS1 and even stronger to its substrate protein Gαi in vitro. NADase activity of PTS1 in vitro was not inhibited by α-defensin-1 in the absence of Gαi. Taken together, these results suggest that α-defensin-1 inhibits PT mainly by inhibiting enzyme activity of PTS1, whereas α-defensin-5 mainly inhibits cellular uptake of PT. These findings will pave the way for optimization of α-defensins as novel therapeutics against whooping cough.
Mismatch Repair Protein Msh2 Is Necessary for Macronuclear Stability and Micronuclear Division in Tetrahymena thermophila
Authors: Wang Lin , Xue Yuhuan , Yang Sitong , Bo Tao , Xu Jing , Wang Wei
Published: June 2023

Mismatch repair (MMR) is a conserved mechanism that is primarily responsible for the repair of DNA mismatches during DNA replication. Msh2 forms MutS heterodimer complexes that initiate the MMR in eukaryotes. The function of Msh2 is less clear under different chromatin structures. Tetrahymena thermophila contains a transcriptionally active macronucleus (MAC) and a transcriptionally silent micronucleus (MIC) in the same cytoplasm. Msh2 is localized in the MAC and MIC during vegetative growth. Msh2 is localized in the perinuclear region around the MIC and forms a spindle-like structure as the MIC divides. During the early conjugation stage, Msh2 is localized in the MIC and disappears from the parental MAC. Msh2 is localized in the new MAC and new MIC during the late conjugation stage. Msh2 also forms a spindle-like structure with a meiotic MIC and mitotic gametic nucleus. MSH2 knockdown inhibits the division of MAC and MIC during vegetative growth and affects cellular proliferation. MSH2 knockdown mutants are sensitive to cisplatin treatment. MSH2 knockdown also affects micronuclear meiosis and gametogenesis during sexual development. Furthermore, Msh2 interacts with MMR-dependent and MMR-independent factors. Therefore, Msh2 is necessary for macronuclear stability, as well as micronuclear mitosis and meiosis in Tetrahymena.
Small Open Reading Frame-Encoded Micro-Peptides: An Emerging Protein World
Authors: Dong Xiaoping , Zhang Kun , Xun Chengfeng , Chu Tianqi , Liang Songping , Zeng Yong , Liu Zhonghua
Published: June 2023

Small open reading frames (sORFs) are often overlooked features in genomes. In the past, they were labeled as noncoding or “transcriptional noise”. However, accumulating evidence from recent years suggests that sORFs may be transcribed and translated to produce sORF-encoded polypeptides (SEPs) with less than 100 amino acids. The vigorous development of computational algorithms, ribosome profiling, and peptidome has facilitated the prediction and identification of many new SEPs. These SEPs were revealed to be involved in a wide range of basic biological processes, such as gene expression regulation, embryonic development, cellular metabolism, inflammation, and even carcinogenesis. To effectively understand the potential biological functions of SEPs, we discuss the history and development of the newly emerging research on sORFs and SEPs. In particular, we review a range of recently discovered bioinformatics tools for identifying, predicting, and validating SEPs as well as a variety of biochemical experiments for characterizing SEP functions. Lastly, this review underlines the challenges and future directions in identifying and validating sORFs and their encoded micropeptides, providing a significant reference for upcoming research on sORF-encoded peptides.
The Cuprizone Mouse Model: A Comparative Study of Cuprizone Formulations from Different Manufacturers
Authors: Beecken Malena , Baumann Louise , Vankriekelsvenne Elise , Manzhula Katerina , Greiner Theresa , Heinig Leo , Schauerte Steffen , Kipp Markus , Joost Sarah
Published: June 2023

The Cuprizone mouse model is widely used in studies on de- and remyelination. In the hands of different experimenters, the Cuprizone concentrations that lead to comparable levels of demyelination differ considerably. The reasons for this variability are unknown. In this study, we tested whether different Cuprizone formulations from different vendors and manufacturers influenced Cuprizone-induced histopathological hallmarks. We intoxicated male C57BL/6 mice with six Cuprizone powders that differed in their manufacturer, vendor, and purity. After five weeks, we analyzed the body weight changes over the course of the experiment, as well as the demyelination, astrogliosis, microgliosis and axonal damage by histological LFB-PAS staining and immunohistochemical labelling of PLP, IBA1, GFAP and APP. All Cuprizone formulations induced demyelination, astrogliosis, microgliosis, axonal damage and a moderate drop in body weight at the beginning of the intoxication period. In a cumulative evaluation of all analyses, two Cuprizone formulations performed weaker than the other formulations. In conclusion, all tested formulations did work, but the choice of Cuprizone formulation may have been responsible for the considerable variability in the experimental outcomes.
A Phase Ib Study of Durvalumab (MEDI4736) in Combination with Carbon-Ion Radiotherapy and Weekly Cisplatin for Patients with Locally Advanced Cervical Cancer (DECISION Study): The Early Safety and Efficacy Results
Authors: Okonogi Noriyuki , Murata Kazutoshi , Yamada Shigeru , Habu Yuji , Hori Makoto , Kurokawa Tomoya , Inaba Yosuke , Fujiwara Tadami , Fujii Yasuhisa , Hanawa Michiko , Kawasaki Yohei , Hattori Yoko , Suzuki Kazuko , Tsuyuki Kyoko , Wakatsuki Masaru , Koto Masashi , Hasegawa Sumitaka , Ishikawa Hitoshi , Hanaoka Hideki , Shozu Makio , Tsuji Hiroshi , Usui Hirokazu
Published: June 2023

We conducted a phase Ib study to examine the safety of a combination of carbon-ion RT (CIRT) with durvalumab (MEDI4736; AstraZeneca) in patients with locally advanced cervical cancer. This was an open-label, single-arm study with a modified 3 + 3 design. Patients with newly diagnosed histologically proven locally advanced cervical cancer were enrolled. All patients received 74.4 Gy of CIRT in 20 fractions and concurrent weekly cisplatin (chemo-CIRT) at a dose of 40 mg/m2. Durvalumab was administered (1500 mg/body) at weeks two and six. The primary endpoint was the incidence of adverse events (AEs) and serious AEs (SAEs), including dose-limiting toxicity (DLT). All three enrolled patients completed the treatment without interruption. One patient developed hypothyroidism after treatment and was determined to be an SAE. No other SAEs were observed. The patient recovered after levothyroxine sodium hydrate treatment. None of the AEs, including hypothyroidism, were associated with DLT in the present study. All three patients achieved complete responses within the CIRT region concerning treatment efficacy. This phase 1b trial demonstrates the safety of combining chemo-CIRT and durvalumab for locally advanced cervical cancer in the early phase. Further research is required as only three patients were included in this study.
Impaired Regulation by IL-35 in Systemic Sclerosis
Authors: Osuna-Gómez Rubén , Castellví Ivan , Mulet Maria , Ortiz Mª Àngels , Brough Douglas E. , Sabzevari Helen , Semnani Roshanak T. , Vidal Silvia
Published: June 2023

This study investigated the role of IL-35 in systemic sclerosis (SSc) patients, focusing on CD4+ T cell response and immunomodulatory cytokine production. By comparing the cytokine levels in healthy donors (HD) and SSc patients using ELISAs, we found a significantly lower plasma IL-35 concentration in the SSc patients (52.1 ± 5.6 vs. 143 ± 11.1, p < 0.001). Notably, the IL-35 levels showed a negative correlation with TGF-β (p < 0.001) and IL-17 (p = 0.04). Assessing the IL-35R expression across cell types in the SSc patients and HDs via flow cytometry, we found higher levels on monocytes (40.7 + 5.7 vs. 20.3 ± 1.9, p < 0.001) and lower levels on CD8+ T cells (61.8 ± 9.2 vs. 83.4 ± 0.8, p < 0.05) in the SSc patients. The addition of recombinant IL-35 to stimulated peripheral blood mononuclear cells reduced the IL-17+CD4+ T cell percentage (9.0 ± 1.5 vs. 4.8 ± 0.7, p < 0.05) and increased the IL-35+CD4+ T percentage (4.1 ± 2.3 vs. 10.2 ± 0.8, p < 0.001). In a Treg:Tresponder cell Sco-culture assay with HD and SSc samples, rIL35 decreased the cell proliferation and levels of IL-17A (178.2 ± 30.5 pg/mL vs. 37.4 ± 6.4 pg/mL, p < 0.001) and TGF-β (4194 ± 777 pg/mL vs. 2413 ± 608 pg/mL, p < 0.01). Furthermore, we observed a positive correlation between the modified Rodnan skin score (mRSS) and TGF-β (p < 0.001), while there was a negative correlation between mRSS and IL-35 (p = 0.004). Interestingly, higher levels of plasmatic IL-35 were detected in individuals with limited disease compared to those with diffuse disease (60.1 ± 8.0 vs. 832.3 ± 4.1, p < 0.05). These findings suggest that IL-35 exhibits anti-inflammatory properties in SSc and it may serve as a marker for disease severity and a therapeutic target.
Proof-of-Concept Human Organ-on-Chip Study: First Step of Platform to Assess Neuro-Immunological Communication Involved in Inflammatory Bowel Diseases
Authors: Gabriel-Segard Tristan , Rontard Jessica , Miny Louise , Dubuisson Louise , Batut Aurélie , Debis Delphine , Gleyzes Mélanie , François Fabien , Larramendy Florian , Soriano Alessandra , Honegger Thibault , Paul Stéphane
Published: June 2023

Inflammatory bowel diseases (IBD) are complex chronic inflammatory disorders of the gastrointestinal (GI) tract. Recent evidence suggests that the gut-brain axis may be pivotal in gastrointestinal and neurological diseases, especially IBD. Here, we present the first proof of concept for a microfluidic technology to model bilateral neuro-immunological communication. We designed a device composed of three compartments with an asymmetric channel that allows the isolation of soma and neurites thanks to microchannels and creates an in vitro synaptic compartment. Human-induced pluripotent stem cell-derived cortical glutamatergic neurons were maintained in soma compartments for up to 21 days. We performed a localized addition of dendritic cells (MoDCs) to either the soma or synaptic compartment. The microfluidic device was coupled with microelectrode arrays (MEAs) to assess the impact on the electrophysiological activity of neurons while adding dendritic cells. Our data highlight that an electrophysiologic signal is transmitted between two compartments of glutamatergic neurons linked by synapses in a bottom-up way when soma is exposed to primed dendritic cells. In conclusion, our study authenticates communication between dendritic cells and neurons in inflammatory conditions such as IBD. This platform opens the way to complexification with gut components to reach a device for pharmacological compound screening by blocking the gut-brain axis at a mucosal level and may help patients.
BDNF and Cognitive Function in Chilean Schizophrenic Patients
Authors: Nieto Rodrigo R. , Silva Hernán , Armijo Alejandra , Nachar Rubén , González Alfonso , Castañeda Carmen Paz , Montes Cristián , Kukuljan Manuel
Published: June 2023

Despite cognitive symptoms being very important in schizophrenia, not every schizophrenic patient has a significant cognitive deficit. The molecular mechanisms underlying the different degrees of cognitive functioning in schizophrenic patients are not sufficiently understood. We studied the relation between brain-derived neurotrophic factor (BDNF) and cognitive functioning in two groups of schizophrenic patients with different cognitive statuses. According to the Montreal Cognitive Assessment (MoCA) results, the schizophrenic patients were classified into two subgroups: normal cognition (26 or more) and cognitive deficit (25 or less). We measured their plasma BDNF levels using ELISAs. The statistical analyses were performed using Spearman’s Rho and Kruskal–Wallis tests. We found a statistically significant positive correlation between the plasma BDNF levels and MoCA score (p = 0.04) in the subgroup of schizophrenic patients with a cognitive deficit (n = 29). However, this correlation was not observed in the patients with normal cognition (n = 11) and was not observed in the total patient group (n = 40). These results support a significant role for BDNF in the cognitive functioning of schizophrenics with some degree of cognitive deficit, but suggest that BDNF may not be crucial in patients with a normal cognitive status. These findings provide information about the molecular basis underlying cognitive deficits in this illness.
Bioactive Glass Modified with Zirconium Incorporation for Dental Implant Applications: Fabrication, Structural, Electrical, and Biological Analysis
Authors: Hammami Imen , Gavinho Sílvia Rodrigues , Pádua Ana Sofia , Sá-Nogueira Isabel , Silva Jorge Carvalho , Borges João Paulo , Valente Manuel Almeida , Graça Manuel Pedro Fernandes
Published: June 2023

Implantology is crucial for restoring aesthetics and masticatory function in oral rehabilitation. Despite its advantages, certain issues, such as bacterial infection, may still arise that hinder osseointegration and result in implant rejection. This work aims to address these challenges by developing a biomaterial for dental implant coating based on 45S5 Bioglass® modified by zirconium insertion. The structural characterization of the glasses, by XRD, showed that the introduction of zirconium in the Bioglass network at a concentration higher than 2 mol% promotes phase separation, with crystal phase formation. Impedance spectroscopy was used, in the frequency range of 102–106 Hz and the temperature range of 200–400 K, to investigate the electrical properties of these Bioglasses, due to their ability to store electrical charges and therefore enhance the osseointegration capacity. The electrical study showed that the presence of crystal phases, in the glass ceramic with 8 mol% of zirconium, led to a significant increase in conductivity. In terms of biological properties, the Bioglasses exhibited an antibacterial effect against Gram-positive and Gram-negative bacteria and did not show cytotoxicity for the Saos-2 cell line at extract concentrations up to 25 mg/mL. Furthermore, the results of the bioactivity test revealed that within 24 h, a CaP-rich layer began to form on the surface of all the samples. According to our results, the incorporation of 2 mol% of ZrO2 into the Bioglass significantly improves its potential as a coating material for dental implants, enhancing both its antibacterial and osteointegration properties.