bluuebunny/crossref_metadata_2025 huggingface dataset:
Developing and Implementing an Action Plan among the “Orang Asali” Minority in Southernmost Thailand for Equitable Accessibility to Public Health Care and Public Services Following the United Nations Sustainable Development Goals
Authors: Meedsen Praves , Sathirapanya Chutarat
Published: March 2023

Ending social inequality by 2030 is a goal of the United Nations’ endorsed sustainable development agenda. Minority or marginalized people are susceptible to social inequality. This action research qualitatively evaluated the requirements for and barriers to full access to public services of the Orang Asali (OA), a minority people living in the Narathiwas province in southernmost Thailand. With the cooperation of the staff of the Southern Border Provinces Administrative Center (SBPAC), we interviewed the OA, local governmental officers and Thai community leaders regarding the OA’s living conditions and health status. Then, an action plan was developed and implemented to raise their living standards with minimal disruption to their traditional cultural beliefs and lifestyle. For systematic follow-ups, a Thai nationality registration process was carried out before the assistance was provided. Living conditions and livelihood opportunities, health care and education were the main targets of the action plan. Universal health coverage (UHC), according to Thai health policy, was applied to OA for holistic health care. The OA were satisfied with the assistance provided to them. While filling the gap of social inequality for the OA is urgent, a balance between the modern and traditional living styles should be carefully considered.
Understanding the Relationship between Urban Biophysical Composition and Land Surface Temperature in a Hot Desert Megacity (Saudi Arabia)
Authors: Addas Abdullah
Published: March 2023

The deteriorations of the thermal environment due to extreme land surface temperature (LST) has become one of the most serious environmental problems in urban areas. The spatial distribution of urban biophysical composition (UBC) has a significant impact on the LST. Therefore, it is essential to understand the relationship between LST and biophysical physical composition (BPC) to mitigate the effects of UHIs. In this study, an attempt was made to understand the relationship between LST and BPC in a hot desert coastal megacity (Jeddah megacity) in Saudi Arabia. Principal component analysis (PCA) was used to understand the factors affecting LST based on remote sensing indices. Correlation and regression analyses were carried out to understand the relationship between LST and BPC and the impact of BPC on LST. The results showed that, in Jeddah city from 2000 to 2021, there was a substantial increase in the built-up area, which increased from 3085 to 5557.98 hectares. Impervious surfaces had a significant impact on the LST, and green infrastructure (GI) was negatively correlated with LST. Based on the PCA results, we found that the GI was a significant factor affecting the LST in Jeddah megacity. The findings of this study, though not contributing to further understanding of the impact of BPC on LST, will provide planners and policy makers with a foundation for developing very effective strategies to improve the eco-environmental quality of Jeddah megacity.
Scanning Electron Microscope Examination as an Alternative to Classical Microbiology in the Diagnostics of Catheter-Related Sepsis?
Authors: Kluzik Anna , Tomczak Hanna , Nowicki Marek , Grześkowiak Małgorzata , Kusza Krzysztof
Published: March 2023

Central venous catheters are essential elements enabling the treatment of intensive care unit patients. However, these catheters are sometimes colonised by both bacteria and fungi, and thus, they may become a potential source of systemic infections—catheter-related bloodstream infections (CRBSI). The identification of the pathogen responsible for CRBSI is a time-consuming process. At the same time, the relationship between the quick identification of the pathogen and the implementation of targeted antibiotic therapy is of key importance for controlling the clinical symptoms of sepsis and septic shock in the patient. Quick diagnosis is of key importance to reduce morbidity and mortality in this group of patients. In our study, we attempted to create a catalogue of images of the most commonly cultured pathogens responsible for CRBSI. An FEI Quanta 250 FEG Scanning Electron Microscope (SEM) was used for measurements. SEM images obtained during the analysis were included in this study. Images of SEM are three-dimensional and comparable to the images seen with the human eye and are a tool used for research and measurement whenever it is necessary to analyse the state of the surface and assess its morphology. The method described in our study will not replace the current procedures recognised as the gold standard, i.e., pathogen culturing, determination of the count of microorganisms (CFU –colony forming units), and assessment of drug sensitivity. However, in some cases, the solution proposed in our study may aid the diagnosis of patients with suspected catheter-related bloodstream infections leading to sepsis and septic shock.
Realfood and Cancer: Analysis of the Reliability and Quality of YouTube Content
Authors: Segado-Fernández Sergio , Herrera-Peco Ivan , Jiménez-Gómez Beatriz , Ruiz Núñez Carlos , Jiménez-Hidalgo Pedro Jesús , Benítez de Gracia Elvira , González-Rodríguez Liliana G. , Torres-Ramírez Cristina , Lozano-Estevan María del Carmen
Published: March 2023

This study analyzes the quality and reliability of videos related to nutrition and cancer on YouTube. Study Design: An observational, retrospective, cross-sectional, time-limited study analyzing activity on the social network YouTube was proposed. Methods: The information from the videos was extracted through an API search tool, using the NodeXL software. The criteria to select the videos on YouTube were the keywords “real food”, “realfood”, and “cancer” and the hashtags #realfood and #cancer were present, videos in English and videos available on 1 December 2022. Results: The DISCERN value in the total number of videos viewed was 2.25 (±0.88) points, indicating low reliability. The videos uploaded by HRU represented only 20.8%. Videos suggesting that the use of foods defined as “real food” could cure cancer without the intervention of any other treatment accounted for 12.5%. Videos that provided external links to scientific/technical evidence verifying the information represented only 13.89% of the total number of videos. Of these videos, 70% corresponded to HRU. The DISCERN value for videos from HRU users was 3.05 (0.88), a value that reflects a good reliability of videos from these users. Conclusions: This study provides information on the content and quality of the videos that we can find on YouTube. We found videos of non-health users who do not base their content on any scientific evidence, with the danger that this entails for the population, but it also highlights that the videos published by HRU have greater reliability and quality, being better perceived by the population, so it is important to encourage healthcare professionals and health institutions to share verified information on YouTube.
The Impact of Medical Nutrition Intervention on the Management of Hyperphosphatemia in Hemodialysis Patients with Stage 5 Chronic Kidney Disease: A Case Series
Authors: Moroșan Elena , Popovici Violeta , Elian Viviana , Dărăban Adriana Maria , Rusu Andreea Ioana , Licu Monica , Mititelu Magdalena , Karampelas Oana
Published: March 2023

The treatment and interdisciplinary management of patients with chronic kidney disease (CKD) continue to improve long-term outcomes. The medical nutrition intervention’s role is to establish a healthy diet plan for kidney protection, reach blood pressure and blood glucose goals, and prevent or delay health problems caused by kidney disease. Our study aims to report the effects of medical nutrition therapy—substituting foods rich in phosphorus-containing additives with ones low in phosphates content on phosphatemia and phosphate binders drug prescription in stage 5 CKD patients with hemodialysis. Thus, 18 adults with high phosphatemia levels (over 5.5 mg/dL) were monitored at a single center. Everyone received standard personalized diets to replace processed foods with phosphorus additives according to their comorbidities and treatment with prosphate binder drugs. Clinical laboratory data, including dialysis protocol, calcemia, and phosphatemia, were evaluated at the beginning of the study, after 30 and 60 days. A food survey was assessed at baseline and after 60 days. The results did not show significant differences between serum phosphate levels between the first and second measurements; thus, the phosphate binders’ initial doses did not change. After 2 months, phosphate levels decreased considerably (from 7.322 mg/dL to 5.368 mg/dL); therefore, phosphate binder doses were diminished. In conclusion, medical nutrition intervention in patients with hemodialysis significantly reduced serum phosphate concentrations after 60 days. Restricting the intake of processed foods containing phosphorus additives—in particularized diets adapted to each patient’s comorbidities—and receiving phosphate binders represented substantial steps to decrease phosphatemia levels. The best results were significantly associated with life expectancy; at the same time, they showed a negative correlation with the dialysis period and participants’ age.
Mouse Models of Mineral Bone Disorders Associated with Chronic Kidney Disease
Authors: Zaloszyc Ariane , Bernardor Julie , Bacchetta Justine , Laverny Gilles , Schmitt Claus Peter
Published: March 2023

Patients with chronic kidney disease (CKD) inevitably develop mineral and bone disorders (CKD–MBD), which negatively impact their survival and quality of life. For a better understanding of underlying pathophysiology and identification of novel therapeutic approaches, mouse models are essential. CKD can be induced by surgical reduction of a functional kidney mass, by nephrotoxic compounds and by genetic engineering specifically interfering with kidney development. These models develop a large range of bone diseases, recapitulating different types of human CKD–MBD and associated sequelae, including vascular calcifications. Bones are usually studied by quantitative histomorphometry, immunohistochemistry and micro-CT, but alternative strategies have emerged, such as longitudinal in vivo osteoblast activity quantification by tracer scintigraphy. The results gained from the CKD–MBD mouse models are consistent with clinical observations and have provided significant knowledge on specific pathomechanisms, bone properties and potential novel therapeutic strategies. This review discusses available mouse models to study bone disease in CKD.
A New Signature That Predicts Progression-Free Survival of Clear Cell Renal Cell Carcinoma with Anti-PD-1 Therapy
Authors: Lin Jingwei , Cai Yingxin , Ma Yuxiang , Pan Jinyou , Wang Zuomin , Zhang Jianpeng , Liu Yangzhou , Zhao Zhigang
Published: March 2023

Immunotherapy has greatly improved the survival time and quality of life of patients with renal cell carcinoma, but the benefits are limited to a small portion of patients. There are too few new biomarkers that can be used to identify molecular subtypes of renal clear cell carcinoma and predict survival time with anti-PD-1 treatment. Single-cell RNA data of clear cell renal cell carcinoma (ccRCC) treated with anti-PD-1 were obtained from public databases, then 27,707 high-quality CD4 + T and CD8 + T cells were obtained for subsequent analysis. Firstly, genes set variation analysis and CellChat algorithm were used to explore potential molecular pathway differences and intercellular communication between the responder and non-responder groups. Additionally, differentially expressed genes (DEGs) between the responder and non-responder groups were obtained using the “edgeR” package, and ccRCC samples from TCGA-KIRC (n = 533) and ICGA-KIRC (n = 91) were analyzed by the unsupervised clustering algorithm to recognize molecular subtypes with different immune characteristics. Finally, using univariate Cox analysis, least absolute shrinkage and selection operator (Lasso) regression, and multivariate Cox regression, the prognosis model of immunotherapy was established and verified to predict the progression-free survival of ccRCC patients treated with anti-PD-1. At the single cell level, there are different signal pathways and cell communication between the immunotherapy responder and non-responder groups. In addition, our research also confirms that the expression level of PDCD1/PD-1 is not an effective marker for predicting the response to immune checkpoint inhibitors (ICIs). The new prognostic immune signature (PIS) enabled the classification of ccRCC patients with anti-PD-1 therapy into high- and low-risk groups, and the progression-free survival times (PFS) and immunotherapy responses were significantly different between these two groups. In the training group, the area under the ROC curve (AUC) for predicting 1-, 2- and 3-year progression-free survival was 0.940 (95% CI: 0.894–0.985), 0.981 (95% CI: 0.960–1.000), and 0.969 (95% CI: 0.937–1.000), respectively. Validation sets confirm the robustness of the signature. This study revealed the heterogeneity between the anti-PD-1 responder and non-responder groups from different angles and established a robust PIS to predict the progression-free survival of ccRCC patients receiving immune checkpoint inhibitors.
CCP1, a Regulator of Tubulin Post-Translational Modifications, Potentially Plays an Essential Role in Cerebellar Development
Authors: Pang Bo , Araki Asuka , Zhou Li , Takebayashi Hirohide , Harada Takayuki , Kadota Kyuichi
Published: March 2023

The cytosolic carboxypeptidase (CCP) 1 protein, encoded by CCP1, is expressed in cerebellar Purkinje cells (PCs). The dysfunction of CCP1 protein (caused by CCP1 point mutation) and the deletion of CCP1 protein (caused by CCP1 gene knockout) all lead to the degeneration of cerebellar PCs, which leads to cerebellar ataxia. Thus, two CCP1 mutants (i.e., Ataxia and Male Sterility [AMS] mice and Nna1 knockout [KO] mice) are used as disease models. We investigated the cerebellar CCP1 distribution in wild-type (WT), AMS and Nna1 KO mice on postnatal days (P) 7–28 to investigate the differential effects of CCP protein deficiency and disorder on cerebellar development. Immunohistochemical and immunofluorescence studies revealed significant differences in the cerebellar CCP1 expression in WT and mutant mice of P7 and P15, but no significant difference between AMS and Nna1 KO mice. Electron microscopy showed slight abnormality in the nuclear membrane structure of PCs in the AMS and Nna1 KO mice at P15 and significant abnormality with depolymerization and fragmentation of microtubule structure at P21. Using two CCP1 mutant mice strains, we revealed the morphological changes of PCs at postnatal stages and indicated that CCP1 played an important role in cerebellar development, most likely via polyglutamylation.
Inter- and Intramolecular RNA–RNA Interactions Modulate the Regulation of Translation Mediated by the 3′ UTR in West Nile Virus
Authors: Romero-López Cristina , Roda-Herreros Margarita , Berzal-Herranz Beatriz , Ramos-Lorente Sara Esther , Berzal-Herranz Alfredo
Published: March 2023

RNA viruses rely on genomic structural elements to accomplish the functions necessary to complete the viral cycle. These elements participate in a dynamic network of RNA–RNA interactions that determine the overall folding of the RNA genome and may be responsible for the fine regulation of viral replication and translation as well as the transition between them. The genomes of members of the genus Flavivirus are characterized by a complexly folded 3′ UTR with a number of RNA structural elements that are conserved across isolates of each species. The present work provides evidence of intra- and intermolecular RNA–RNA interactions involving RNA structural elements in the 3′ UTR of the West Nile virus genome. The intermolecular interactions can be visualized in vitro by the formation of molecular dimers involving the participation of at least the SLI and 3′DB elements. Certainly, the 3′ UTR of dengue virus, which lacks the SLI element, forms molecular dimers in lower quantities via a single interaction site, probably 3′DB. The functional analysis of sequence or deletion mutants revealed an inverse relationship between 3′ UTR dimerization and viral translation efficiency in cell cultures. A network of RNA–RNA interactions involving 3′ UTR structural elements might therefore exist, helping to regulate viral translation.
Production of Silver Nano-Inks and Surface Coatings for Anti-Microbial Food Packaging and Its Ecological Impact
Authors: Manikandan N. Arul , McCann Ronan , Kakavas Dimitrios , Rochfort Keith D. , Sreenilayam Sithara P. , Alkan Godze , Stornetta Tom , McGivern Allan Robert , Grintzalis Konstantinos , Friedrich Bernd , Foley Greg , Brabazon Dermot , Freeland Brian
Published: March 2023

Food spoilage is an ongoing global issue that contributes to rising carbon dioxide emissions and increased demand for food processing. This work developed anti-bacterial coatings utilising inkjet printing of silver nano-inks onto food-grade polymer packaging, with the potential to enhance food safety and reduce food spoilage. Silver nano-inks were synthesised via laser ablation synthesis in solution (LaSiS) and ultrasound pyrolysis (USP). The silver nanoparticles (AgNPs) produced using LaSiS and USP were characterised using transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectroscopy, UV-Vis spectrophotometry and dynamic light scattering (DLS) analysis. The laser ablation technique, operated under recirculation mode, produced nanoparticles with a small size distribution with an average diameter ranging from 7–30 nm. Silver nano-ink was synthesised by blending isopropanol with nanoparticles dispersed in deionised water. The silver nano-inks were printed on plasma-cleaned cyclo-olefin polymer. Irrespective of the production methods, all silver nanoparticles exhibited strong antibacterial activity against E. coli with a zone of inhibition exceeding 6 mm. Furthermore, silver nano-inks printed cyclo-olefin polymer reduced the bacterial cell population from 1235 (±45) × 106 cell/mL to 960 (±110) × 106 cell/mL. The bactericidal performance of silver-coated polymer was comparable to that of the penicillin-coated polymer, wherein a reduction in bacterial population from 1235 (±45) × 106 cell/mL to 830 (±70) × 106 cell/mL was observed. Finally, the ecotoxicity of the silver nano-ink printed cyclo-olefin polymer was tested with daphniids, a species of water flea, to simulate the release of coated packaging into a freshwater environment.
Xerostomia and Its Cellular Targets
Authors: Kim Yoon-Jung
Published: March 2023

Xerostomia, the subjective feeling of a dry mouth associated with dysfunction of the salivary glands, is mainly caused by radiation and chemotherapy, various systemic and autoimmune diseases, and drugs. As saliva plays numerous essential roles in oral and systemic health, xerostomia significantly reduces quality of life, but its prevalence is increasing. Salivation mainly depends on parasympathetic and sympathetic nerves, and the salivary glands responsible for this secretion move fluid unidirectionally through structural features such as the polarity of acinar cells. Saliva secretion is initiated by the binding of released neurotransmitters from nerves to specific G-protein-coupled receptors (GPCRs) on acinar cells. This signal induces two intracellular calcium (Ca2+) pathways (Ca2+ release from the endoplasmic reticulum and Ca2+ influx across the plasma membrane), and this increased intracellular Ca2+ concentration ([Ca2+]i) causes the translocation of the water channel aquaporin 5 (AQP5) to the apical membrane. Consequently, the GPCR-mediated increased [Ca2+]i in acinar cells promotes saliva secretion, and this saliva moves into the oral cavity through the ducts. In this review, we seek to elucidate the potential of GPCRs, the inositol 1,4,5-trisphosphate receptor (IP3R), store-operated Ca2+ entry (SOCE), and AQP5, which are essential for salivation, as cellular targets in the etiology of xerostomia.
Organophosphate-Pesticide-Mediated Immune Response Modulation in Invertebrates and Vertebrates
Authors: Bernal-González Karime Guadalupe , Covantes-Rosales Carlos Eduardo , Camacho-Pérez Milton Rafael , Mercado-Salgado Ulises , Barajas-Carrillo Victor Wagner , Girón-Pérez Daniel Alberto , Montoya-Hidalgo Ashley Carolina , Díaz-Resendiz Karina Janice Guadalupe , Barcelos-García Rocío Guadalupe , Toledo-Ibarra Gladys Alejandra , Girón-Pérez Manuel Iván
Published: March 2023

Organophosphate pesticides (OPs) have greatly facilitated food production worldwide, and their use is not limited to agriculture and the control of pests and disease vectors. However, these substances can directly affect the immune response of non-target organisms. In this sense, exposure to OPs can have negative effects on innate and adaptive immunity, promoting deregulation in humoral and cellular processes such as phagocytosis, cytokine expression, antibody production, cell proliferation, and differentiation, which are crucial mechanisms for host defense against external agents. This review focuses on the scientific evidence of exposure to OPs and their toxic effects on the immune system of non-target organisms (invertebrates and vertebrates) from a descriptive perspective of the immuno-toxic mechanisms associated with susceptibility to the development of bacterial, viral, and fungal infectious diseases. During the exhaustive review, we found that there is an important gap in the study of non-target organisms, examples of which are echinoderms and chondrichthyans. It is therefore important to increase the number of studies on other species directly or indirectly affected by Ops, to assess the degree of impact at the individual level and how this affects higher levels, such as populations and ecosystems.
FapydG in the Shadow of OXOdG—A Theoretical Study of Clustered DNA Lesions
Authors: Karwowski Bolesław T.
Published: March 2023

Genetic information, irrespective of cell type (normal or cancerous), is exposed to a range of harmful factors, which can lead to more than 80 different types of DNA damage. Of these, oxoG and FapyG have been identified as the most abundant in normoxic and hypoxic conditions, respectively. This article considers d[AFapyGAOXOGA]*[TCTCT] (oligo-FapyG) with clustered DNA lesions (CDLs) containing both the above types of damage at the M06-2x/6-31++G** level of theory in the condensed phase. Furthermore, the electronic properties of oligo-FapyG were analysed in both equilibrated and non-equilibrated solvation–solute interaction modes. The vertical/adiabatic ionization potential (VIP, AIP) and electron affinity (VEA, AEA) of the investigated ds-oligo were found as follows in [eV]: 5.87/5.39 and −1.41/−2.09, respectively. The optimization of the four ds-DNA spatial geometries revealed that the transFapydG was energetically privileged. Additionally, CDLs were found to have little influence on the ds-oligo structure. Furthermore, for the FapyGC base-pair isolated from the discussed ds-oligo, the ionization potential and electron affinity values were higher than those assigned to OXOGC. Finally, a comparison of the influence of FapyGC and OXOGC on charge transfer revealed that, in contrast to the OXOGC base-pair, which, as expected, acted as a radical cation/anion sink in the oligo-FapyG structure, FapyGC did not significantly affect charge transfer (electron–hole and excess–electron). The results presented below indicate that 7,8-dihydro-8-oxo-2′-deoxyguanosine plays a significant role in charge transfer through ds-DNA containing CDL and indirectly has an influence on the DNA lesion recognition and repair process. In contrast, the electronic properties obtained for 2,6-diamino-4-hydroxy-5-foramido-2′deoxypyrimidine were found to be too weak to compete with OXOG to influence charge transfer through the discussed ds-DNA containing CDL. Because increases in multi-damage site formation are observed during radio- or chemotherapy, understanding their role in the above processes can be crucial for the efficiency and safety of medical cancer treatment.
Schiedeella bajaverapacensis (Orchidaceae, Spiranthinae), a New Orchid Species from Guatemala
Authors: Archila Morales Fredy L. , Lipińska Monika M. , Dudek Magdalena , Szlachetko Dariusz L.
Published: March 2023

Guatemala is recognized for its diverse and rich flora and fauna. It is estimated that over 1200 orchid species, classified in 223 genera, are known to occur in this rather small, yet megadiverse country. While studying the diversity of this plant group in the department of Baja Verapaz, we found individuals that clearly belonged to the genus Schiedeella, but whose features did not fit any previously known species. At that time, nine terrestrial taxon representatives were known to occur in Guatemala. We conducted the morphological analysis in accordance with the standard procedures of classical taxonomy. For phylogenetic reconstruction, 59 sequences of the ITS region and 48 of the trnL-trnF marker were applied. The topology of trees was obtained based on the Bayesian inference. Schiedeella bajaverapacensis was described and illustrated based on morphological evidence, and its taxonomic position was confirmed by phylogenetic analyses. The new entity is the 10th Schiedeella representative known from Guatemala.
The Regulatory Roles of Ezh2 in Response to Lipopolysaccharide (LPS) in Macrophages and Mice with Conditional Ezh2 Deletion with LysM-Cre System
Authors: Kunanopparat Areerat , Leelahavanichkul Asada , Visitchanakun Peerapat , Kueanjinda Patipark , Phuengmaung Pornpimol , Sae-khow Kritsanawan , Boonmee Atsadang , Benjaskulluecha Salisa , Palaga Tanapat , Hirankarn Nattiya
Published: March 2023

The responses of macrophages to lipopolysaccharide (LPS) might determine the direction of clinical manifestations of sepsis, which is the immune response against severe infection. Meanwhile, the enhancer of zeste homologue 2 (Ezh2), a histone lysine methyltransferase of epigenetic regulation, might interfere with LPS response. Transcriptomic analysis on LPS-activated wild-type macrophages demonstrated an alteration of several epigenetic enzymes. Although the Ezh2-silencing macrophages (RAW264.7), using small interfering RNA (siRNA), indicated a non-different response to the control cells after a single LPS stimulation, the Ezh2-reducing cells demonstrated a less severe LPS tolerance, after two LPS stimulations, as determined by the higher supernatant TNF-α. With a single LPS stimulation, Ezh2 null (Ezh2flox/flox; LysM-Crecre/−) macrophages demonstrated lower supernatant TNF-α than Ezh2 control (Ezh2fl/fl; LysM-Cre−/−), perhaps due to an upregulation of Socs3, which is a suppressor of cytokine signaling 3, due to the loss of the Ezh2 gene. In LPS tolerance, Ezh2 null macrophages indicated higher supernatant TNF-α and IL-6 than the control, supporting an impact of the loss of the Ezh2 inhibitory gene. In parallel, Ezh2 null mice demonstrated lower serum TNF-α and IL-6 than the control mice after an LPS injection, indicating a less severe LPS-induced hyper-inflammation in Ezh2 null mice. On the other hand, there were similar serum cytokines after LPS tolerance and the non-reduction of serum cytokines after the second dose of LPS, indicating less severe LPS tolerance in Ezh2 null mice compared with control mice. In conclusion, an absence of Ezh2 in macrophages resulted in less severe LPS-induced inflammation, as indicated by low serum cytokines, with less severe LPS tolerance, as demonstrated by higher cytokine production, partly through the upregulated Socs3.
Models of Congenital Adrenal Hyperplasia for Gene Therapies Testing
Authors: Glazova Olga , Bastrich Asya , Deviatkin Andrei , Onyanov Nikita , Kaziakhmedova Samira , Shevkova Liudmila , Sakr Nawar , Petrova Daria , Vorontsova Maria V. , Volchkov Pavel
Published: March 2023

The adrenal glands are important endocrine organs that play a major role in the stress response. Some adrenal glands abnormalities are treated with hormone replacement therapy, which does not address physiological requirements. Modern technologies make it possible to develop gene therapy drugs that can completely cure diseases caused by mutations in specific genes. Congenital adrenal hyperplasia (CAH) is an example of such a potentially treatable monogenic disease. CAH is an autosomal recessive inherited disease with an overall incidence of 1:9500–1:20,000 newborns. To date, there are several promising drugs for CAH gene therapy. At the same time, it remains unclear how new approaches can be tested, as there are no models for this disease. The present review focuses on modern models for inherited adrenal gland insufficiency and their detailed characterization. In addition, the advantages and disadvantages of various pathological models are discussed, and ways of further development are suggested.
Proanthocyanidins: Impact on Gut Microbiota and Intestinal Action Mechanisms in the Prevention and Treatment of Metabolic Syndrome
Authors: Redondo-Castillejo Rocío , Garcimartín Alba , Hernández-Martín Marina , López-Oliva María Elvira , Bocanegra Aránzazu , Macho-González Adrián , Bastida Sara , Benedí Juana , Sánchez-Muniz Francisco J.
Published: March 2023

The metabolic syndrome (MS) is a cluster of risk factors, such as central obesity, hyperglycemia, dyslipidemia, and arterial hypertension, which increase the probability of causing premature mortality. The consumption of high-fat diets (HFD), normally referred to high-saturated fat diets, is a major driver of the rising incidence of MS. In fact, the altered interplay between HFD, microbiome, and the intestinal barrier is being considered as a possible origin of MS. Consumption of proanthocyanidins (PAs) has a beneficial effect against the metabolic disturbances in MS. However, there are no conclusive results in the literature about the efficacy of PAs in improving MS. This review allows a comprehensive validation of the diverse effects of the PAs on the intestinal dysfunction in HFD-induced MS, differentiating between preventive and therapeutic actions. Special emphasis is placed on the impact of PAs on the gut microbiota, providing a system to facilitate comparison between the studies. PAs can modulate the microbiome toward a healthy profile and strength barrier integrity. Nevertheless, to date, published clinical trials to verify preclinical findings are scarce. Finally, the preventive consumption of PAs in MS-associated dysbiosis and intestinal dysfunction induced by HFD seems more successful than the treatment strategy.
Pro-Arrhythmic Potential of Accumulated Uremic Toxins Is Mediated via Vulnerability of Action Potential Repolarization
Authors: van Ham Willem B. , Cornelissen Carlijn M. , Polyakova Elizaveta , van der Voorn Stephanie M. , Ligtermoet Merel L. , Monshouwer-Kloots Jantine , Vos Marc A. , Bossu Alexandre , van Rooij Eva , van der Heyden Marcel A. G. , van Veen Toon A. B.
Published: March 2023

Chronic kidney disease (CKD) is represented by a diminished filtration capacity of the kidneys. End-stage renal disease patients need dialysis treatment to remove waste and toxins from the circulation. However, endogenously produced uremic toxins (UTs) cannot always be filtered during dialysis. UTs are among the CKD-related factors that have been linked to maladaptive and pathophysiological remodeling of the heart. Importantly, 50% of the deaths in dialysis patients are cardiovascular related, with sudden cardiac death predominating. However, the mechanisms responsible remain poorly understood. The current study aimed to assess the vulnerability of action potential repolarization caused by exposure to pre-identified UTs at clinically relevant concentrations. We exposed human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and HEK293 chronically (48 h) to the UTs indoxyl sulfate, kynurenine, or kynurenic acid. We used optical and manual electrophysiological techniques to assess action potential duration (APD) in the hiPSC-CMs and recorded IKr currents in stably transfected HEK293 cells (HEK-hERG). Molecular analysis of KV11.1, the ion channel responsible for IKr, was performed to further understand the potential mechanism underlying the effects of the UTs. Chronic exposure to the UTs resulted in significant APD prolongation. Subsequent assessment of the repolarization current IKr, often most sensitive and responsible for APD alterations, showed decreased current densities after chronic exposure to the UTs. This outcome was supported by lowered protein levels of KV11.1. Finally, treatment with an activator of the IKr current, LUF7244, could reverse the APD prolongation, indicating the potential modulation of electrophysiological effects caused by these UTs. This study highlights the pro-arrhythmogenic potential of UTs and reveals a mode of action by which they affect cardiac repolarization.
The Endothelial Glycocalyx as a Target of Excess Soluble Fms-like Tyrosine Kinase-1
Authors: Schulz Annika , Drost Carolin C. , Hesse Bettina , Beul Katrin , Boeckel Göran R. , Lukasz Alexander , Pavenstädt Hermann , Brand Marcus , Di Marco Giovana S.
Published: March 2023

Soluble fms-like tyrosine kinase-1 (sFlt-1) is a secreted protein that binds heparan sulfate expressed on the endothelial glycocalyx (eGC). In this paper we analyze how excess sFlt-1 causes conformational changes in the eGC, leading to monocyte adhesion, a key event triggering vascular dysfunction. In vitro exposure of primary human umbilical vein endothelial cells to excess sFlt-1 decreased eGC height and increased stiffness as determined by atomic force microscopy (AFM). Yet, structural loss of the eGC components was not observed, as indicated by Ulex europaeus agglutinin I and wheat germ agglutinin staining. Moreover, the conformation observed under excess sFlt-1, a collapsed eGC, is flat and stiff with unchanged coverage and sustained content. Functionally, this conformation increased the endothelial adhesiveness to THP-1 monocytes by about 35%. Heparin blocked all these effects, but the vascular endothelial growth factor did not. In vivo administration of sFlt-1 in mice also resulted in the collapse of the eGC in isolated aorta analyzed ex vivo by AFM. Our findings show that excess sFlt-1 causes the collapse of the eGC and favors leukocyte adhesion. This study provides an additional mechanism of action by which sFlt-1 may cause endothelial dysfunction and injury.
MicroRNA as a Diagnostic Tool, Therapeutic Target and Potential Biomarker in Cutaneous Malignant Melanoma Detection—Narrative Review
Authors: Poniewierska-Baran Agata , Zadroga Łukasz , Danilyan Edo , Małkowska Paulina , Niedźwiedzka-Rystwej Paulina , Pawlik Andrzej
Published: March 2023

Melanoma is the most serious type of skin cancer, causing a large majority of deaths but accounting for only ~1% of all skin cancer cases. The worldwide incidence of malignant melanoma is increasing, causing a serious socio-economic problem. Melanoma is diagnosed mainly in young and middle-aged people, which distinguishes it from other solid tumors detected mainly in mature people. The early detection of cutaneous malignant melanoma (CMM) remains a priority and it is a key factor limiting mortality. Doctors and scientists around the world want to improve the quality of diagnosis and treatment, and are constantly looking for new, promising opportunities, including the use of microRNAs (miRNAs), to fight melanoma cancer. This article reviews miRNA as a potential biomarker and diagnostics tool as a therapeutic drugs in CMM treatment. We also present a review of the current clinical trials being carried out worldwide, in which miRNAs are a target for melanoma treatment.
Theoretical Study on the Gas-Phase and Aqueous Interface Reaction Mechanism of Criegee Intermediates with 2-Methylglyceric Acid and the Nucleation of Products
Authors: Li Lei , Zhang Qingzhu , Wei Yuanyuan , Wang Qiao , Wang Wenxing
Published: March 2023

Criegee intermediates (CIs) are important in the sink of many atmospheric substances, including alcohols, organic acids, amines, etc. In this work, the density functional theory (DFT) method was used to calculate the energy barriers for the reactions of CH3CHOO with 2-methyl glyceric acid (MGA) and to evaluate the interaction of the three functional groups of MGA. The results show that the reactions involving the COOH group of MGA are negligibly affected, and that hydrogen bonding can affect the reactions involving α-OH and β-OH groups. The water molecule has a negative effect on the reactions of the COOH group. It decreases the energy barriers of reactions involving the α-OH and β-OH groups as a catalyst. The Born-Oppenheimer molecular dynamic (BOMD) was applied to simulate the reactions of CH3CHOO with MGA at the gas-liquid interface. Water molecule plays the role of proton transfer in the reaction. Gas-phase calculations and gas-liquid interface simulations demonstrate that the reaction of CH3CHOO with the COOH group is the main pathway in the atmosphere. The molecular dynamic (MD) simulations suggest that the reaction products can form clusters in the atmosphere to participate in the formation of particles.
Glycosylation Analysis of Urinary Peptidome Highlights IGF2 Glycopeptides in Association with CKD
Authors: Lohia Sonnal , Latosinska Agnieszka , Zoidakis Jerome , Makridakis Manousos , Mischak Harald , Glorieux Griet , Vlahou Antonia , Jankowski Vera
Published: March 2023

Chronic kidney disease (CKD) is prevalent in 10% of world’s adult population. The role of protein glycosylation in causal mechanisms of CKD progression is largely unknown. The aim of this study was to identify urinary O-linked glycopeptides in association to CKD for better characterization of CKD molecular manifestations. Urine samples from eight CKD and two healthy subjects were analyzed by CE-MS/MS and glycopeptides were identified by a specific software followed by manual inspection of the spectra. Distribution of the identified glycopeptides and their correlation with Age, eGFR and Albuminuria were evaluated in 3810 existing datasets. In total, 17 O-linked glycopeptides from 7 different proteins were identified, derived primarily from Insulin-like growth factor-II (IGF2). Glycosylation occurred at the surface exposed IGF2 Threonine 96 position. Three glycopeptides (DVStPPTVLPDNFPRYPVGKF, DVStPPTVLPDNFPRYPVG and DVStPPTVLPDNFPRYP) exhibited positive correlation with Age. The IGF2 glycopeptide (tPPTVLPDNFPRYP) showed a strong negative association with eGFR. These results suggest that with aging and deteriorating kidney function, alterations in IGF2 proteoforms take place, which may reflect changes in mature IGF2 protein. Further experiments corroborated this hypothesis as IGF2 increased plasma levels were observed in CKD patients. Protease predictions, considering also available transcriptomics data, suggest activation of cathepsin S with CKD, meriting further investigation.
Comprehensive Analysis of Universal Stress Protein Family Genes and Their Expression in Fusarium oxysporum Response of Populus davidiana × P. alba var. pyramidalis Louche Based on the Transcriptome
Authors: Diao Jian , Gu Wei , Jiang Zhehui , Wang Jiaqi , Zou Hongfei , Zong Cheng , Ma Ling
Published: March 2023

Universal stress proteins (USPs) are typical stress-inducible proteins that function directly in a variety of biotic or abiotic stresses and effectively protect plants from complex, adverse environments. However, the expression patterns of USP genes under pathogen stress and their molecular mechanisms in stress resistance have not been reported in detail. In this study, 46 USP genes were identified from Populus trichocarpa (PtrUSPs), and their biological characteristics were comprehensively analyzed based on phylogeny, physicochemical properties of proteins, and gene structures. The promoter regions of PtrUSPs contain a variety of cis-acting elements related to hormone and stress response. The results of a collinearity analysis showed that PtsrUSPs were highly conserved with homologous genes from four other representative species (Arabidopsis thaliana, Eucalyptus grandis, Glycine max, and Solanum lycopersicum). Furthermore, RNA-Seq analysis showed that the expression of 46 USPs from P. davidiana × P. alba var. pyramidalis Louche (PdpapUSPs) was significantly induced by Fusarium oxysporum. The co-expression network and gene ontology analysis of PtrUSPs showed that they participated in the response to stress and response to stimulus through precise coordination. The results of this paper systematically revealed the biological characteristics of PtrUSPs and the characteristics of their response to F. oxysporum stress, which will lay a theoretical foundation for improving genetic traits and the breeding of poplar disease-resistant varieties in subsequent studies.
Molecular Aspects Implicated in Dantrolene Selectivity with Respect to Ryanodine Receptor Isoforms
Authors: Gaburjakova Jana , Gaburjakova Marta
Published: March 2023

Dantrolene is an intra-cellularly acting skeletal muscle relaxant used for the treatment of the rare genetic disorder, malignant hyperthermia (MH). In most cases, MH susceptibility is caused by dysfunction of the skeletal ryanodine receptor (RyR1) harboring one of nearly 230 single-point MH mutations. The therapeutic effect of dantrolene is the result of a direct inhibitory action on the RyR1 channel, thus suppressing aberrant Ca2+ release from the sarcoplasmic reticulum. Despite the almost identical dantrolene-binding sequence exits in all three mammalian RyR isoforms, dantrolene appears to be an isoform-selective inhibitor. Whereas RyR1 and RyR3 channels are competent to bind dantrolene, the RyR2 channel, predominantly expressed in the heart, is unresponsive. However, a large body of evidence suggests that the RyR2 channel becomes sensitive to dantrolene-mediated inhibition under certain pathological conditions. Although a consistent picture of the dantrolene effect emerges from in vivo studies, in vitro results are often contradictory. Hence, our goal in this perspective is to provide the best possible clues to the molecular mechanism of dantrolene’s action on RyR isoforms by identifying and discussing potential sources of conflicting results, mainly coming from cell-free experiments. Moreover, we propose that, specifically in the case of the RyR2 channel, its phosphorylation could be implicated in acquiring the channel responsiveness to dantrolene inhibition, interpreting functional findings in the structural context.
Development of Highly Sensitive Digital Droplet PCR for Detection of cKIT Mutations in Circulating Free DNA That Mediate Resistance to TKI Treatment for Gastrointestinal Stromal Tumor (GIST)
Authors: Rassner Michael , Waldeck Silvia , Follo Marie , Jilg Stefanie , Philipp Ulrike , Jolic Martina , Wehrle Julius , Jost Philipp J. , Peschel Christian , Illert Anna Lena , Duyster Justus , Scherer Florian , von Bubnoff Nikolas
Published: March 2023

Background: Mutations in cKIT or PDGFRA are found in up to 90% of patients with gastrointestinal stromal tumors (GISTs). Previously, we described the design, validation, and clinical performance of a digital droplet (dd)PCR assay panel for the detection of imatinib-sensitive cKIT and PDFGRA mutations in circulating tumor (ct)DNA. In this study, we developed and validated a set of ddPCR assays for the detection of cKIT mutations mediating resistance to cKIT kinase inhibitors in ctDNA. In addition, we cross-validated these assays using next generation sequencing (NGS). Methods: We designed and validated five new ddPCR assays to cover the most frequent cKIT mutations mediating imatinib resistance in GISTs. For the most abundant imatinib-resistance-mediating mutations in exon 17, a drop-off, probe-based assay was designed. Dilution series (of decreasing mutant (MUT) allele frequency spiked into wildtype DNA) were conducted to determine the limit of detection (LoD). Empty controls, single wildtype controls, and samples from healthy individuals were tested to assess specificity and limit of blank (LoB). For clinical validation, we measured cKIT mutations in three patients and validated results using NGS. Results: Technical validation demonstrated good analytical sensitivity, with a LoD ranging between 0.006% and 0.16% and a LoB ranging from 2.5 to 6.7 MUT fragments/mL. When the ddPCR assays were applied to three patients, the abundance of ctDNA in serial plasma samples reflected the individual disease course, detected disease activity, and indicated resistance mutations before imaging indicated progression. Digital droplet PCR showed good correlation to NGS for individual mutations, with a higher sensitivity of detection. Conclusions: This set of ddPCR assays, together with our previous set of cKIT and PDGFRA mutations assays, allows for dynamic monitoring of cKIT and PDGFRA mutations during treatment. Together with NGS, the GIST ddPCR panel will complement imaging of GISTs for early response evaluation and early detection of relapse, and thus it might facilitate personalized decision-making.
Advantages of Understanding the Molecular Mechanisms of Angiogenesis in Various Physiological and Pathological Conditions
Authors: Elmoselhi Adel B.
Published: March 2023

The aim of this Special Issue is to highlight the diverse benefits and approaches to studying angiogenesis in various physiological and pathological conditions, such as damaged tissues, impaired embryonic development, cancer progression, and cardiovascular and chronic inflammatory disorders [...]
Chitosan Composites with Bacterial Cellulose Nanofibers Doped with Nanosized Cerium Oxide: Characterization and Cytocompatibility Evaluation
Authors: Petrova Valentina A. , Gofman Iosif V. , Dubashynskaya Natallia V. , Golovkin Alexey S. , Mishanin Alexander I. , Ivan’kova Elena M. , Romanov Dmitry P. , Khripunov Albert K. , Vlasova Elena N. , Migunova Alexandra V. , Baranchikov Alexander E. , Ivanov Vladimir K. , Yakimansky Alexander V. , Skorik Yury A.
Published: March 2023

In this work, new composite films were prepared by incorporating the disintegrated bacterial cellulose (BCd) nanofibers and cerium oxide nanoparticles into chitosan (CS) matrices. The influence of the amount of nanofillers on the structure and properties of the polymer composites and the specific features of the intermolecular interactions in the materials were determined. An increase in film stiffness was observed as a result of reinforcing the CS matrix with BCd nanofibers: the Young’s modulus increased from 4.55 to 6.3 GPa with the introduction of 5% BCd. A further increase in Young’s modulus of 6.7 GPa and a significant increase in film strength (22% increase in yield stress compared to the CS film) were observed when the BCd concentration was increased to 20%. The amount of nanosized ceria affected the structure of the composite, followed by a change in the hydrophilic properties and texture of the composite films. Increasing the amount of nanoceria to 8% significantly improved the biocompatibility of the films and their adhesion to the culture of mesenchymal stem cells. The obtained nanocomposite films combine a number of favorable properties (good mechanical strength in dry and swollen states, improved biocompatibility in relation to the culture of mesenchymal stem cells), which allows us to recommend them for use as a matrix material for the culture of mesenchymal stem cells and wound dressings.