bluuebunny/crossref_metadata_2025 huggingface dataset:
Evolution of Fentanyl Prescription Patterns and Administration Routes in Primary Care in Salamanca, Spain: A Comprehensive Analysis from 2011 to 2022
Authors: Torres-Bueno Cristina , Sanchez-Barba Mercedes , Miron-Canelo Jose-Antonio , Gonzalez-Nunez Veronica
Published: August 2024

(1) Background: The escalating use of opioids contributes to social, health, and economic crises. In Spain, a notable surge in the medical prescription of opioids in recent years has been observed. The aim of this work was to assess the consumption rate of fentanyl, categorised by the different administration routes, in Primary Care in the province of Salamanca (Spain) spanning the years 2011 to 2022, and to compare it with the national trend and with data from the US. (2) Methods: Doses per inhabitant per day (DHD) were calculated, and interannual variations, as well as consumption rates, were subject to thorough analysis. (3) Results: The prevalence of fentanyl use in Salamanca has doubled from 1.21 DHD in 2011 to 2.56 DHD in 2022, with the transdermal system (TD) as the predominant administration route. This upward trajectory mirrors the national trend, yet the rise in fentanyl use is markedly lower than the reported data in the US. This finding may be attributed to an ageing population and potentially inappropriate fentanyl prescriptions, i.e., for the management of chronic non-cancer pain and other off-label prescriptions. (4) Conclusions: The use of fentanyl in Salamanca, particularly through transdermal systems, doubled from 2011 to 2022, aligning with the national trend. Preventive measures are imperative to prevent fentanyl misuse and moderate the observed escalation in consumption rates.
Wall Drawing #736: Revealing Sol LeWitt’s Ink Mural Technique Using a Multi-Analytical Approach
Authors: Ferretti Adele , Degano Ilaria , Filomena Marta , La Nasa Jacopo , Campanella Beatrice , Legnaioli Stefano , Penoni Sara , Pintus Renata , Todaro Cristiana , Modugno Francesca
Published: August 2024

Sol LeWitt, a pioneer of conceptual art, created during his career over 1350 wall drawings, including the Wall Drawing #736 (1993) at the Center for Contemporary Art Luigi Pecci in Prato (Prato, Italy). The painting, executed by Andrea Marescalchi and Antony Sansotta under LeWitt’s instructions, features a grid of coloured rectangles obtained by overlapping different layers of inks. During a 2021 restoration by the Wall Paintings and Stuccoes Department of the Opificio delle Pietre Dure (Firenze, Italy), an in-depth investigation of the composition and the materials used by LeWitt’s assistants in producing Wall Drawing #736 was performed. A multi-analytical approach entailing Raman spectroscopy, high-performance liquid chromatography coupled to diode array and high-resolution mass spectrometry, gas chromatography–mass spectrometry (GC–MS), and pyrolysis coupled with GC–MS was applied. Our results revealed the use of animal glue, shellac resin, paraffin wax, linseed oil, and various organic pigments. The binder in the preparation layer was identified as poly(vinyl acetate), while poly(n-butyl methacrylate) was determined as a fixative. This research provided valuable insights into LeWitt’s techniques. The acquired knowledge on the paint technique is highly relevant in supporting conservators in restoration and consolidating the many wall drawings produced exploiting the same technique all over the world.
Rural Landscapes as Cultural Heritage and Identity along a Romanian River
Authors: Dragan Alexandru , Creţan Remus , Jucu Ioan Sebastian , Oancea Oana Andreea
Published: August 2024

In contemporary narratives, rural landscapes and identities, as judged by the inhabitants’ sense of belonging, continue to be important points that need to be (re)considered when discussing places as instruments for local development. This paper pushes forward these debates and explores the construction of rural and regional identity in Romania with the help of a case study that focuses on perceptions of rural landscapes as cultural heritage in two rural communities situated on opposite sides of the river Mureș. Based on participant and ethnographic observation and making use of a survey and a qualitative analysis, this paper highlights the most significant features of the local cultural identities and their rural landscape heritage expressed by the two sets of people, which often coincide with the most important characteristics attributed to the regions separated by the river Mureș. The results of this study underline the dividing role of the river in the preservation of rural identity and cultural heritage. They also demonstrate that rural landscapes and the construction of identities along the river have generated both development opportunities and various risks.
Nutrient Dynamics and Resource-Use Efficiency in Greenhouse Strawberries: Effects of Control Variables in Closed-Loop Hydroponics
Authors: Lim Mi Young , Kim So Hui , Roh Mi Young , Choi Gyeong Lee , Kim Dongpil
Published: August 2024

The importance of implementing recirculating drainage for greenhouse strawberries is often overlooked because of the low electrical conductivity (EC) of drainage and transpiration despite the large area of cultivation in Korea. In this study, we analyzed the growth of strawberry crops and their water and individual nutrient use efficiency when using closed-loop hydroponics in greenhouses. The study consisted of two parts: Experiment (Exp) 1 and Exp 2, each of which employed a different closed-loop hydroponic control method. In Exp 1, the system was controlled solely based on the EC of the drainage mixed with raw water. In Exp 2, the nutrient solution (NS) was corrected according to the ion concentrations in the drainage, with correction intervals of 2 weeks, 4 weeks, and no correction, while the control treatment was open-loop. In Exp 1, a prolonged increase in drainage EC resulted in an imbalance in NS ion composition and reduced fruit yield. In Exp 2, the NS composition was stabilized through periodic nutrient correction, and the fruit yield of the closed-loop treatments did not differ from the open-loop treatment. However, the closed-loop with 2- and 4-week correction in Exp 2 showed 94% and 88% higher nutrient use efficiency (NUE), respectively, than an open-loop system. Among the closed-loop treatments, the 2- and 4-week correction intervals had 36% and 32% higher NUE than the no-correction treatment. Furthermore, the 2-week correction interval showed a 3% improvement in NUE compared to the 4-week interval. These findings highlight the importance of considering a shorter ion correction interval for optimal NUE and normal plant growth in recirculating hydroponic systems of strawberry cultivation.
1-Methylcyclopropene and Bacillus Treatment Affects Blueberry Storage Quality and Antioxidant Levels
Authors: Wu Shuangshuang , Zhang Yan , Wang Mengyang , Liu Yahui , Li Lingxiang , He Feng , Xiao Jiaxin
Published: August 2024

Blueberries are rich in a variety of functional substances and have high nutritional and health values, but they are not resistant to decline during fresh storage. Here, the effects of six Bacillus species on the storage quality of, and antioxidant levels in, the southern highbush blueberry ‘O’Neal’ fruit were investigated. Bacillus treatments reduced the fruit decay rate, slowed fruit quality decline, inhibited malondialdehyde accumulation, and increased superoxide dismutase and peroxidase activity levels. Bacillus altitudinis Y-14 had the best effect overall. Furthermore, the effects of 1-methylcyclopropene (1-MCP), B. altitudinis Y-14, and 1-MCP + B. altitudinis Y-14 treatments on the storage quality and antioxidant of rabbiteye blueberry ‘Brightwell’ fruit were investigated, and each treatment effectively reduced the decay rate and weight loss of fruit. When stored for 25 days, the decay rate of the 1-MCP + B. altitudinis Y-14 group was only 8.33%, significantly lower than that of the control. The three treatments delayed the decline in fruit quality, inhibited malondialdehyde accumulation, and increased superoxide dismutase and peroxidase activity levels. The 1-MCP + B. altitudinis Y-14 treatment was more conducive to prolonging the postharvest storage period of blueberries and had the best effect in delaying the decline in fruit quality. Thus, combined 1-MCP and B. altitudinis Y-14 treatment may be an effective way to improve the storage quality and extend the storage period of blueberries, which provides a new way for storing and transporting blueberries to reduce costs and improve economic benefits.
An Evaluation of Different Sweet Olive Cultivars with Different Ripening Degrees Grown in the Puglia Region, Southeastern Italy
Authors: Alhajj Ali Salem , Mazzeo Andrea , Trani Antonio , Pitardi Simona , Bisceglie Sara , Ferrara Giuseppe
Published: August 2024

Some olive cultivars grown in southeastern Italy are characterized by the production of olives with a reduced level of bitterness. They are known as sweet olive cultivars and fruits are usually consumed directly or cooked without any debittering process, offering either health benefits to consumers, thanks to the high content of antioxidants, or an economic benefit to farmers for their higher price with respect to both table and oil olives. This study evaluates and compares the organoleptic, pomological, chemical, and physical parameters of seven sweet olive cultivars at different ripening degrees in the Puglia region over 8 weeks of maturity stage for two consecutive seasons (2022 and 2023). The organoleptic evaluation was performed by a restricted panel of usual consumers/experts of sweet olives. The results showed a higher preference for the olive cultivars locally named Triggiano Dolce, Cerasella, and Mele. Significant differences in weight, length, and width of the fruits were observed based on both cultivar and year. The phenolic composition of olive cultivars was significantly affected by both cultivar and harvest year, with Cazzinicchio and Cellina di Nardò having the highest total polyphenols. The analysis of water fraction extracted from olive samples by liquid chromatography coupled with mass spectrometry led to the identification of eleven compounds belonging to the secoiroids, phenylpropanoids, phenylethanolids, and flavonoids classes. The comparison of these compounds among the studied cultivars highlighted significant differences.
Biomass-to-Green Hydrogen: A Review of Techno-Economic-Enviro Assessment of Various Production Methods
Authors: Ghasemi Amir , Nikafshan Rad Hima , Akrami Mohammad
Published: August 2024

H2 is considered a practical substitute for fossil fuels, especially for transportation by road and air, created either from fossil fuels or through the process of electrolysis of water. Research questions were included based on numerous research and the analysis of articles. The cost analysis of H2 processes, techno-economic hurdles in commercialization, and the economic comparison of various H2-production methods were the basis for the study of papers. The current research examines the different methods of thermochemical, biological, and electrochemical processes utilized in converting biomass into hydrogen. The benefits, constraints, and significant enhancements of every procedure are outlined. The examination assesses the cost of production, the level of technology readiness, and the potential for scalability. Thermochemical techniques, such as gasification and steam reforming, are effective at producing hydrogen. Steam gasification is perfect for moist and dry biomass in the absence of an oxidizing agent. Dark fermentation is more efficient for biological conversion because it requires less energy. Moreover, the electrochemical procedure is viable for biomass. Thermochemical treatment is significantly more advanced than biological or electrochemical treatment when it comes to scaling opportunities based on comparisons of current processes. The results of this research show that biomass–hydrogen processes have the potential for increasing H2 production, but further enhancements are needed to produce larger quantities for competitiveness.
Under-Five Mortality and Associated Risk Factors in Children Hospitalized at David Bernardino Pediatric Hospital (DBPH), Angola: A Hierarchical Approach
Authors: Avelino Israel C. , Van-Dúnem Joaquim , Varandas Luís
Published: August 2024

Reducing under-five mortality is a crucial indicator of overall development in a country. However, in Angola, understanding the factors contributing to hospital deaths in this vulnerable demographic remains incomplete despite improvements in healthcare infrastructure and public health policies. With one of the highest under-five mortality rates in sub-Saharan Africa, Angola faces significant challenges such as malaria, malnutrition, pneumonia, neonatal conditions, and intestinal infectious diseases, which are the leading causes of death among children. This study aimed to identify factors associated with hospital deaths among children aged 28 days to five years admitted to DBPH in Luanda between May 2022 and June 2023. Using a hospital-based case-control design, the study included 1020 children, among whom 340 experienced hospital deaths. Distal and intermediate determinants emerged as primary predictors of hospital mortality, showing significant associations with: mother without schooling (OR [95%CI] 4.3 [1.2–15.7], p < 0.027); frequent alcohol consumption during pregnancy (OR [95%CI] 3.8 [2.5–5.9], p < 0.001); hospital stay ≤24 h (OR [95%CI] 13.8 [6.2–30.8], p < 0.001); poor nutritional status (OR [95%CI] 2.1 [1.4–3.2], p < 0.001); short interbirth interval (OR [95%CI] 1.7 [1.1–2.5], p < 0.014); maternal age ≤19 years (OR [95%CI] 5.6 [3.0–10.8], p < 0.001); and maternal age ≥35 years (OR [95%CI] 2.1 [1.2–3.7], p < 0.006). These findings highlight the preventable nature of most under-five hospital deaths and underscore the urgent need to address social inequities and improve the quality of primary healthcare services to effectively reduce child mortality in Angola.
Co-Producing Paws on Campus: A Psychoeducational Dog-Facilitated Programme for University Students Experiencing Mental Health Difficulties
Authors: Williams Joanne M. , Bradfield Jillian , Gardiner Andrew , Pendry Patricia , Wauthier Laura
Published: August 2024

Declining student mental health is a global public health issue. Campus-based animal-assisted interventions (AAIs) are popular and effective interventions to prevent and alleviate symptoms. How to design, implement and evaluate evidence-based, student-centred interventions that enjoy sustained stakeholder buy-in and support is less known. This paper presents the procedures and results of a three-stage co-production method and the resulting curriculum of a novel AAI aimed at university students experiencing serious mental health problems. Stage 1 shaped the focus and structure of the intervention based on online student surveying (N = 204) and consultations with stakeholders (N = 10), including representatives of Student Well-being Services leadership, veterinarians, animal welfare charities and Therapets volunteers. In Stage 2, we conducted co-production workshops with post-graduate students (N = 6), developing the curriculum based on Stage 1 insights. In Stage 3, through iterative prototyping and student feedback (N = 22) the Paws on Campus programme was finalised, resulting in a series of four, one-hour themed sessions: (1) Thoughts and Feelings, (2) Well-being and Welfare, (3) Care and Compassion and (4) Problem Solving and Help Seeking. We describe the co-production method and resulting programme characteristics and provide considerations for others interested in developing effective and sustainable AAIs for their respective populations and contexts.
Doxycycline-Mediated Control of Cyclin D2 Overexpression in Human-Induced Pluripotent Stem Cells
Authors: Qiao Aijun , Wei Yuhua , Liu Yanwen , Kahn-Krell Asher , Ye Lei , Nguyen Thanh , Zhang Jianyi
Published: August 2024

Previous studies have demonstrated that when the cyclin D2 (CCND2), a cell-cycle regulatory protein, is overexpressed in human-induced pluripotent stem cells (hiPSCs), cardiomyocytes (CMs) differentiated from these CCND2-overexpressing hiPSCs can proliferate after transplantation into infarcted hearts, which significantly improves the cells’ potency for myocardial regeneration. However, persistent CM proliferation could lead to tumor growth or the development of arrhythmogenic complications; thus, the goal of the current study was to generate a line of hiPSCs in which CCND2 overexpression could be tightly controlled. First, we transfected hiPSCs with vectors coding for a doxycycline-inducible Tet-On transactivator and S. pyogenes dCas9 fused to the VPR activation domain; then, the same hiPSCs were engineered to express guide RNAs targeting the CCND2 promotor. Thus, treatment with doxycycline (dox) activated dCas9-VPR expression, and the guide RNAs directed dCas9-VPR to the CCND2 promoter, which activated CCND2 expression. Subsequent experiments confirmed that CCND2 expression was dox-dependent in this newly engineered line of hiPSCs (doxCCND2-hiPSCs): CCND2 protein was abundantly expressed after 48 h of treatment with dox and declined to near baseline level ~96 h after dox treatment was discontinued.
Strong Bases Design: Key Techniques and Stability Issues
Authors: Kulsha Andrey V. , Ivashkevich Oleg A. , Lyakhov Dmitry A. , Michels Dominik
Published: August 2024

Theoretical design of molecular superbases has been attracting researchers for more than twenty years. General approaches were developed to make the bases potentially stronger, but less attention was paid to the stability of the predicted structures. Hence, only a small fraction of the theoretical research has led to positive experimental results. Possible stability issues of extremely strong bases are extensively studied in this work using quantum chemical calculations on a high level of theory. Several step-by-step design examples are discussed in detail, and general recommendations are given to avoid the most common stability problems. New potentially stable structures are theoretically studied to demonstrate the future prospects of molecular superbases design.
Expression Patterns of MOTS-c in Adrenal Tumors: Results from a Preliminary Study
Authors: Kamiński Kacper , Blatkiewicz Małgorzata , Szyszka Marta , Olechnowicz Anna , Komarowska Hanna , Klimont Anna , Wierzbicki Tomasz , Karczewski Marek , Ruchała Marek , Rucinski Marcin
Published: August 2024

Adrenal tumors, such as adrenocortical carcinoma (ACC), adrenocortical adenoma (ACA), and pheochromocytoma (PCC) are complex diseases with unclear causes and treatments. Mitochondria and mitochondrial-derived peptides (MDPs) are crucial for cancer cell survival. The primary aim of this study was to analyze samples from different adrenal diseases, adrenocortical carcinoma, adrenocortical adenoma, and pheochromocytoma, and compare them with normal adrenal tissue to determine whether the expression levels of the mitochondrial open reading frame of the 12S rRNA type-c (MOTS-c) gene and protein vary between different types of adrenal tumors compared to healthy controls using qPCR, ELISA, and IHC methods. Results showed decreased MOTS-c mRNA expression in all adrenal tumors compared to controls, while serum MOTS-c protein levels increased in ACA and PCC but not in ACC. The local distribution of MOTS-c protein in adrenal tissue was reduced in all tumors. Notably, MOTS-c protein expression declined with ACC progression (stages III and IV) but was unrelated to patient age or sex. Tumor size and testosterone levels positively correlated with MOTS-c mRNA but negatively with serum MOTS-c protein. Additionally, serum MOTS-c protein correlated positively with glucose, total cholesterol, HDL, LDL, and SHGB levels. These findings suggest disrupted expression of MOTS-c in the spectrum of adrenal diseases, which might be caused by mechanisms involving increased mitochondrial dysfunction and structural changes in the tissue associated with disease progression. This study provides a detailed examination of MOTS-c mRNA and protein in adrenal tumors, indicating the potential role of MDPs in tumor biology and progression.
Humanized L184Q Mutated Surfactant Protein C Gene Alters Alveolar Type 2 Epithelial Cell Fate
Authors: Jain Krishan G. , Liu Yang , Zhao Runzhen , Muire Preeti J. , Zhang Jiwang , Zang Qun Sophia , Ji Hong-Long
Published: August 2024

Alveolar type 2 epithelial (AT2) cells synthesize surfactant protein C (SPC) and repair an injured alveolar epithelium. A mutated surfactant protein C gene (SftpcL184Q, Gene ID: 6440) in newborns has been associated with respiratory distress syndrome and pulmonary fibrosis. However, the underlying mechanisms causing Sftpc gene mutations to regulate AT2 lineage remain unclear. We utilized three-dimensional (3D) feeder-free AT2 organoids in vitro to simulate the alveolar epithelium and compared AT2 lineage characteristics between WT (C57BL/6) and SftpcL184Q mutant mice using colony formation assays, immunofluorescence, flow cytometry, qRT-PCR, and Western blot assays. The AT2 numbers were reduced significantly in SftpcL184Q mice. Organoid numbers and colony-forming efficiency were significantly attenuated in the 3D cultures of primary SftpcL184Q AT2 cells compared to those of WT mice. Podoplanin (PDPN, Alveolar type 1 cell (AT1) marker) expression and transient cell count was significantly increased in SftpcL184Q organoids compared to in the WT mice. The expression levels of CD74, heat shock protein 90 (HSP90), and ribosomal protein S3A1 (RPS3A1) were not significantly different between WT and SftpcL184Q AT2 cells. This study demonstrated that humanized SftpcL184Q mutation regulates AT2 lineage intrinsically. This regulation is independent of CD74, HSP90, and RPS3A1 pathways.
Insights on the Mechanical Properties of SARS-CoV-2 Particles and the Effects of the Photosensitizer Hypericin
Authors: Mariangeli Matteo , Moreno Ana , Delcanale Pietro , Abbruzzetti Stefania , Diaspro Alberto , Viappiani Cristiano , Bianchini Paolo
Published: August 2024

SARS-CoV-2 is a highly pathogenic virus responsible for the COVID-19 disease. It belongs to the Coronaviridae family, characterized by a phospholipid envelope, which is crucial for viral entry and replication in host cells. Hypericin, a lipophilic, naturally occurring photosensitizer, was reported to effectively inactivate enveloped viruses, including SARS-CoV-2, upon light irradiation. In addition to its photodynamic activity, Hyp was found to exert an antiviral action also in the dark. This study explores the mechanical properties of heat-inactivated SARS-CoV-2 viral particles using Atomic Force Microscopy (AFM). Results reveal a flexible structure under external stress, potentially contributing to the virus pathogenicity. Although the fixation protocol causes damage to some particles, correlation with fluorescence demonstrates colocalization of partially degraded virions with their genome. The impact of hypericin on the mechanical properties of the virus was assessed and found particularly relevant in dark conditions. These preliminary results suggest that hypericin can affect the mechanical properties of the viral envelope, an effect that warrants further investigation in the context of antiviral therapies.
Galectin-2 Agglutinates Helicobacter pylori via Lipopolysaccharide Containing H Type I Under Weakly Acidic Conditions
Authors: Sasaki Takaharu , Oyama Midori , Kubota Mao , Isshiki Yasunori , Takeuchi Tomoharu , Tanaka Toru , Tanikawa Takashi , Tamura Mayumi , Arata Yoichiro , Hatanaka Tomomi
Published: August 2024

Galectins are β-galactoside-binding animal lectins involved in various biological functions, such as host defense. Galectin-2 and -3 are members of the galectin family that are expressed in the stomach, including the gastric mucosa and surface mucous cells. Galectin-3 exhibits aggregation and bactericidal activity against Helicobacter pylori in a β-galactoside-dependent manner. We previously reported that galectin-2 has the same activity under neutral pH conditions. In this study, the H. pylori aggregation activity of galectin-2 was examined under weakly acidic conditions, in which H. pylori survived. Galectin-2 agglutinated H. pylori even at pH 6.0, but not at pH 5.0, correlating with its structural stability, as determined using circular dichroism. Additionally, galectin-2 binding to the lipopolysaccharide (LPS) of H. pylori cultured under weakly acidic conditions was investigated using affinity chromatography and Western blotting. Galectin-2 could bind to H. pylori LPS containing H type I, a Lewis antigen, in a β-galactoside-dependent manner. In contrast, galectin-3 was structurally more stable than galectin-2 under acidic conditions and bound to H. pylori LPS containing H type I and Lewis X. In conclusion, galectin-2 and -3 might function cooperatively in the defense against H. pylori in the stomach under different pH conditions.
Crosstalk between Ethylene, Jasmonate and ABA in Response to Salt Stress during Germination and Early Plant Growth in Cucurbita pepo
Authors: Alonso Sonsoles , Gautam Keshav , Iglesias-Moya Jessica , Martínez Cecilia , Jamilena Manuel
Published: August 2024

The crosstalk of phytohormones in the regulation of growth and development and the response of plants to environmental stresses is a cutting-edge research topic, especially in crop species. In this paper, we study the role and crosstalk between abscisic acid (ABA), ethylene (ET), and jasmonate (JA) in the control of germination and seedling growth in water or in standard nutrient solution and under salt stress (supplemented with 100–200 mM NaCl). The roles of ET and JA were studied using squash ET- and JA-deficient mutants aco1a and lox3a, respectively, while the crosstalk between ET, JA, and ABA was determined by comparing the expression of the key ABA, JA, and ET genes in wild-type (WT) and mutant genotypes under standard conditions and salt stress. Data showed that ET and JA are positive regulators of squash germination, a function that was found to be mediated by downregulating the ABA biosynthesis and signaling pathways. Under salt stress, aco1a germinated earlier than WT, while lox3a showed the same germination rate as WT, indicating that ET, but not JA, restricts squash germination under unfavorable salinity conditions, a function that was also mediated by upregulation of ABA. ET and JA were found to be negative regulators of plant growth during seedling establishment, although ET inhibits both the aerial part and the root, while JA inhibits only the root. Both aco1a and lox3a mutant roots showed increased tolerance to salt stress, a phenotype that was found to be mainly mediated by JA, although we cannot exclude that it is also mediated by ABA.
Steroidomics in Men with Schizophrenia
Authors: Hill Martin , Velíková Marta , Hovorková Tereza , Bulant Josef , Janšáková Katarína , Valeš Karel
Published: August 2024

Schizophrenia is associated with numerous abnormalities, including imbalances in all hormonal axes, among which steroids play a major role. Steroidomic studies therefore represent a promising tool for early diagnosis and appropriate treatment of schizophrenia. A total of 51 adult male schizophrenics aged 27 (22, 34) years (shown as median with quartiles) and 16 healthy controls (HCs) aged 28 (25, 32) years were enrolled into this study. Our results showed the effective differentiation of men with schizophrenia from controls based on steroidomic profiles. We also found an altered metabolic pathway from pregnenolone and its sulfate (PREG/S) to cortisol in schizophrenics with several metabolic bottlenecks such as lower PREG levels due to increased PREG sulfation and/or suppressed PREGS desulfation and attenuated conversion of 17-hydroxy-PREG to 17-hydroxy-progesterone, as well as the results suggestive of suppressed CYP11B1 activity. In contrast, steroid molar ratios suggested two counterregulatory steps involving increased conversion of PREG/S to 17-hydroxy-PREG/S and decreased conversion of cortisol to cortisone, which may maintain unchanged basal cortisol levels but may not ensure a sufficient cortisol response to stress. Our data also indicated a trend to higher 7α-, 7β-, and 16α-hydroxylation that may counteract the autoimmune complications and proinflammatory processes accompanying schizophrenia. Finally, a possible suppression of HSD17B3 activity was suggested, resulting in decreased circulating testosterone levels with increased androstenedione levels.
Genetic Influences of Proinflammatory Cytokines on Pain Severity in Patients with Temporomandibular Disorders
Authors: Zlendić Marko , Vrbanović Đuričić Ema , Gall Trošelj Koraljka , Tomljanović Marko , Vuković Đerfi Kristina , Alajbeg Iva Z.
Published: August 2024

This case-control study investigated single nucleotide polymorphism (SNP) genotypes (CXC motif chemokine ligand 8 (CXCL8): rs2227306 and rs2227307 and tumor necrosis factor (TNF): rs1800629) in 85 patients with pain-related temporomandibular disorders (TMDp) and 85 controls to explore their associations with TMDp presence, pain intensity (low/high), and the presence of chronic arthralgia/myalgia. TMDp was diagnosed using a validated protocol, and polymorphisms were genotyped from buccal mucosa swabs using TaqMan assays. High pain intensity individuals had an increased risk for carrying minor allele “G” (rs2227307) and “T” (rs2227306) compared to controls (76% vs. 55.3%, p = 0.012; 72% vs. 54.1%, p = 0.030, respectively). Carriers of the minor allele “G” (rs2227307) were more prevalent in TMDp patients with arthralgia compared to controls (70.30% vs. 55.30%, p = 0.037). According to logistic regression, the most important predictors for high pain intensity were minor allele “G” of rs2227307 (OR 2.435, 95% CI 1.123–5.282), increasing age (OR 1.038, 95% CI 1.002–1.075), and female sex (OR 4.592, 95% CI 1.289–16.361). The explored gene polymorphisms were not significant risk factors for TMDp presence. These findings highlight the importance of genetic variations, particularly rs2227307, in understanding the diverse clinical manifestations of temporomandibular disorders.
Acetylation of Steroidogenic Acute Regulatory Protein Sensitizes 17β-Estradiol Regulation in Hormone-Sensitive Breast Cancer Cells
Authors: Manna Pulak R. , Molehin Deborah , Ahmed Ahsen U. , Yang Shengping , Reddy P. Hemachandra
Published: August 2024

An imbalance in estrogen signaling is a critical event in breast tumorigenesis. The majority of breast cancers (BCs) are hormone-sensitive; they majorly express the estrogen receptor (ER+) and are activated by 17β-estradiol (E2). The steroidogenic acute regulatory protein (StAR) mediates the rate-limiting step in steroid biosynthesis. The dysregulation of the epigenetic machinery, modulating E2 levels, is a primary occurrence for promoting breast tumorigenesis. StAR expression, concomitant with E2 synthesis, was reported to be aberrantly high in human and mouse hormone-dependent BC cells compared with their non-cancerous counterparts. However, the mechanism of action of StAR remains poorly understood. We discovered StAR as an acetylated protein and have identified a number of lysine (K) residues that are putatively acetylated in malignant and non-malignant breast cells, using LC-MS/MS (liquid chromatography–tandem mass spectrometry), suggesting they differently influence E2 synthesis in mammary tissue. The treatment of hormone-sensitive MCF7 cells with a variety of histone deacetylase inhibitors (HDACIs), at therapeutically and clinically relevant doses, identified a few additional StAR acetylated lysine residues. Among a total of fourteen StAR acetylomes undergoing acetylation and deacetylation, K111 and K253 were frequently recognized either endogenously or in response to HDACIs. Site-directed mutagenesis studies of these two StAR acetylomes, pertaining to K111Q and K253Q acetylation mimetic states, resulted in increases in E2 levels in ER+ MCF7 and triple negative MB-231 BC cells, compared with their values seen with human StAR. Conversely, these cells carrying K111R and K253R deacetylation mutants diminished E2 biosynthesis. These findings provide novel and mechanistic insights into intra-tumoral E2 regulation by elucidating the functional importance of this uncovered StAR post-translational modification (PTM), involving acetylation and deacetylation events, underscoring the potential of StAR as a therapeutic target for hormone-sensitive BC.
Understanding Secondary Sarcopenia Development in Young Adults Using Pig Model with Chronic Pancreatitis
Authors: Tomaszewska Ewa , Wojtysiak Dorota , Grzegorzewska Agnieszka , Świątkiewicz Małgorzata , Donaldson Janine , Arciszewski Marcin B. , Dresler Sławomir , Puzio Iwona , Szymańczyk Sylwia , Dobrowolski Piotr , Bonior Joanna , Mielnik-Błaszczak Maria , Kuc Damian , Muszyński Siemowit
Published: August 2024

Chronic pancreatitis (CP) in young individuals may lead to disease-related secondary sarcopenia (SSARC), characterized by muscle loss and systemic inflammation. In this study, CP was induced in young pigs, and serum levels of key hormones, muscle fiber diameters in various muscles, and the mRNA expression of genes related to oxidative stress and programmed cell death were assessed. A decrease in muscle fiber diameters was observed in SSARC pigs, particularly in the longissimus and diaphragm muscles. Hormonal analysis revealed alterations in dehydroepiandrosterone, testosterone, oxytocin, myostatin, and cortisol levels, indicating a distinct hormonal response in SSARC pigs compared to controls. Oxytocin levels in SSARC pigs were significantly lower and myostatin levels higher. Additionally, changes in the expression of catalase (CAT), caspase 8 (CASP8), B-cell lymphoma 2 (BCL2), and BCL2-associated X protein (BAX) mRNA suggested a downregulation of oxidative stress response and apoptosis regulation. A reduced BAX/BCL2 ratio in SSARC pigs implied potential caspase-independent cell death pathways. The findings highlight the complex interplay between hormonal changes and muscle degradation in SSARC, underscoring the need for further research into the apoptotic and inflammatory pathways involved in muscle changes due to chronic organ inflammation in young individuals.
Proteomic Characterization of Changes in Mouse Brain Cortex Protein Expression at Different Post-Mortem Intervals: A Preliminary Study for Forensic Biomarker Identification
Authors: Bonelli Martina , Di Giuseppe Fabrizio , Tupone Nicola , Di Virgilio Vimal , Catena Antonio Maria , Locatelli Marcello , Ascani Giuliano , Giammaria Gianluigi , Ciccarelli Renata , D’Ovidio Cristian , Angelucci Stefania
Published: August 2024

Accuracy in the evaluation of death-induced tissue degradation for thanato-chronological purposes is strictly dependent on the condition of the biological source as well as on the precision of post-mortem interval (PMI) estimation. Thus, the optimization of tissue handling and identification of sensitive post-mortem biomarkers could help establish a timeline for post-mortem events. To this aim, we investigated the proteome changes in cortex samples of 6-week-old female SAMR1 mice over a post-mortem time course. After death, brain tissue was removed immediately (T0), and after 4, 8, 12, 24, and 32 h, four mice were used for each time period, and animals were maintained at 4 °C until brain removal. Dissected tissues were frozen at −80 °C until processed. Proteomic analysis, performed on samples related to early and late PMIs (<24 h and >24 h post-mortem, respectively) showed protein level changes as compared to T0 samples, with a remarkable increase in Calpain11 in the early PMI, as well as in Caspases 7 and 8 together with Gasdermin 3 in late PMI. These findings were confirmed by LIFT mass spectrometry technology and western blot analysis and, although requiring further investigation in other biological samples, suggest that these proteins could be considered as putative biomarkers of different PMIs.
Genetic, Clinical, Epidemiological, and Immunological Profiling of IgG Response Duration after SARS-CoV-2 Infection
Authors: Póvoa da Costa Flávia , Sarges Kevin Matheus Lima de , Silva Rosilene da , Santos Erika Ferreira dos , Nascimento Matheus Holanda do , Rodrigues Alice Maciel , Cantanhede Marcos Henrique Damasceno , Rodrigues Fabíola Brasil Barbosa , Viana Maria de Nazaré do Socorro de Almeida , Leite Mauro de Meira , Oliveira Camille Ferreira de , Neves Pablo Fabiano Moura das , Pereira Neto Gabriel dos Santos , Brito Mioni Thieli Figueiredo Magalhães de , Silva Andréa Luciana Soares da , Henriques Daniele Freitas , Quaresma Juarez Antônio Simões , Falcão Luiz Fábio Magno , Queiroz Maria Alice Freitas , Vallinoto Izaura Maria Vieira Cayres , Vallinoto Antonio Carlos Rosário , Viana Giselle Maria Rachid , Santos Eduardo José Melo dos
Published: August 2024

The IgG response against SARS-CoV-2 infection can persist for over six months (long response; LR). However, among 30% of those infected, the duration can be as short as three months or less (short response; SR). The present study assembled serological data on the anti-SARS-CoV-2 IgG response duration of two previous studies and integrated these results with the plasmatic cytokine levels and genetic profile of 10 immune-relevant SNPs that were also previously published, along with the plasmatic total IgG, IgA, and IgM levels, allowing for the genetic, clinical, immunological, and epidemiological aspects of the post-COVID-19 IgG response duration to be understood. The SR was associated with previous mild acute COVID-19 and with an SNP (rs2228145) in IL6R related to low gene expression. Additionally, among the SR subgroup, no statistically significant Spearman correlations were observed between the plasma levels of IL-17A and the Th17 regulatory cytokines IFN-γ (rs = 0.2399; p = 0.1043), IL-4 (rs = 0.0273; p = 0.8554), and IL-2 (rs = 0.2204; p = 0.1365), while among the LR subgroup, weaker but statistically significant Spearman correlations were observed between the plasma levels of IL-17A and IFN-γ (rs = 0.3873; p = 0.0016), IL-4 (rs = 0.2671; p = 0.0328), and IL-2 (rs = 0.3959; p = 0.0012). These results suggest that the Th17 response mediated by the IL-6 pathway has a role in the prolonged IgG response to SARS-CoV-2 infection.
ABCC1 Is a ΔNp63 Target Gene Overexpressed in Squamous Cell Carcinoma
Authors: La Banca Veronica , De Domenico Sara , Nicolai Sara , Gatti Veronica , Scalera Stefano , Maugeri Marcello , Mauriello Alessandro , Montanaro Manuela , Pahnke Jens , Candi Eleonora , D’Amico Silvia , Peschiaroli Angelo
Published: August 2024

The transcription factor ΔNp63 plays a pivotal role in maintaining the integrity of stratified epithelial tissues by regulating the expression of distinct target genes involved in lineage specification, cell stemness, cell proliferation and differentiation. Here, we identified the ABC transporter subfamily member ABCC1 as a novel ΔNp63 target gene. We found that in immortalized human keratinocytes and in squamous cell carcinoma (SCC) cells, ∆Np63 induces the expression of ABCC1 by physically occupying a p63-binding site (p63 BS) located in the first intron of the ABCC1 gene locus. In cutaneous SCC and during the activation of the keratinocyte differentiation program, ∆Np63 and ABCC1 levels are positively correlated raising the possibility that ABCC1 might be involved in the regulation of the proliferative/differentiative capabilities of squamous tissue. However, we did not find any gross alteration in the structure and morphology of the epidermis in humanized hABCC1 knock-out mice. Conversely, we found that the genetic ablation of ABCC1 led to a marked reduction in inflammation-mediated proliferation of keratinocytes, suggesting that ABCC1 might be involved in the regulation of keratinocyte proliferation upon inflammatory/proliferative signals. In line with these observations, we found a significant increase in ABCC1 expression in squamous cell carcinomas (SCCs), a tumor type characterized by keratinocyte hyper-proliferation and a pro-inflammatory tumor microenvironment. Collectively, these data uncover ABCC1 as an additional ∆Np63 target gene potentially involved in those skin diseases characterized by dysregulation of proliferation/differentiation balance.
The Human OCTN Sub-Family: Gene and Protein Structure, Expression, and Regulation
Authors: Galluccio Michele , Tripicchio Martina , Pochini Lorena
Published: August 2024

OCTN1 and OCTN2 are membrane transport proteins encoded by the SLC22A4 and SLC22A5 genes, respectively. Even though several transcripts have been predicted by bioinformatics for both genes, only one functional protein isoform has been described for each of them. Both proteins are ubiquitous, and depending on the physiopathological state of the cell, their expression is regulated by well-known transcription factors, although some aspects have been neglected. A plethora of missense variants with uncertain clinical significance are reported both in the dbSNP and the Catalogue of Somatic Mutations in Cancer (COSMIC) databases for both genes. Due to their involvement in human pathologies, such as inflammatory-based diseases (OCTN1/2), systemic primary carnitine deficiency (OCTN2), and drug disposition, it would be interesting to predict the impact of variants on human health from the perspective of precision medicine. Although the lack of a 3D structure for these two transport proteins hampers any speculation on the consequences of the polymorphisms, the already available 3D structures for other members of the SLC22 family may provide powerful tools to perform structure/function studies on WT and mutant proteins.
A Newly Developed Anti-L1CAM Monoclonal Antibody Targets Small Cell Lung Carcinoma Cells
Authors: Yamaguchi Miki , Hirai Sachie , Idogawa Masashi , Sumi Toshiyuki , Uchida Hiroaki , Sakuma Yuji
Published: August 2024

Few effective treatments are available for small cell lung cancer (SCLC), indicating the need to explore new therapeutic options. Here, we focus on an antibody–drug conjugate (ADC) targeting the L1 cell adhesion molecule (L1CAM). Several publicly available databases reveal that (1) L1CAM is expressed at higher levels in SCLC cell lines and tissues than in those of lung adenocarcinoma and (2) the expression levels of L1CAM are slightly higher in SCLC tissues than in adjacent normal tissues. We conducted a series of in vitro experiments using an anti-L1CAM monoclonal antibody (termed HSL175, developed in-house) and the recombinant protein DT3C, which consists of diphtheria toxin lacking the receptor-binding domain but containing the C1, C2, and C3 domains of streptococcal protein G. Our HSL175-DT3C conjugates theoretically kill cells only when the conjugates are internalized by the target (L1CAM-positive) cells through antigen–antibody interaction. The conjugates (an ADC analog) were effective against two SCLC-N (NEUROD1 dominant) cell lines, Lu-135 and STC-1, resulting in decreased viability. In addition, L1CAM silencing rendered the two cell lines resistant to HSL175-DT3C conjugates. These findings suggest that an ADC consisting of a humanized monoclonal antibody based on HSL175 and a potent anticancer drug would be effective against SCLC-N cells.
Benefits and Challenges of Drug-Coated Balloons in Peripheral Artery Disease: From Molecular Mechanisms to Clinical Practice
Authors: Tataru Dan-Alexandru , Lazar Florin-Leontin , Onea Horea-Laurentiu , Homorodean Calin , Ober Mihai-Claudiu , Olinic Maria , Spinu Mihail , Olinic Dan-Mircea
Published: August 2024

Multiple clinical trials have reported favorable outcomes after drug-coated balloon therapy for peripheral artery disease in above-the-knee and below-the-knee lesions and in both de novo and in-stent restenosis. However, there are still insufficient data to identify and tackle the risk factors associated with a higher risk of restenosis, which is the primary concern for patients who are treated with an endovascular approach. A modern armamentarium, which includes improved lesion preparation techniques such as plaque modification balloons, mechanical atherectomy, intravascular lithotripsy, and imaging, is crucial for obtaining better long-term clinical outcomes. Moreover, a better understanding of the molecular properties of drug-coated balloons has led to improved devices that could tackle the shortcomings of previous generations. This comprehensive review focuses on drug-coated balloon technology as a tool to treat peripheral artery disease and the effects of the molecular mechanisms involved in preventing vascular restenosis.
Assessing the Efficacy of Protease Inactivation for the Preservation of Bioactive Amphibian Skin Peptides
Authors: Samgina Tatiana Yu. , Mazur Dmitrii M. , Lebedev Albert T.
Published: August 2024

The skin of amphibians is a rich source of peptides with a wide range of biological activities. They are stored in secretory granules in an inactive form. Upon stimulation, they are secreted together with proteases into the skin. Once activated, they rapidly exert their biological effects, including fighting microorganisms and predators, while their excess is immediately destroyed by the released proteases. To keep bioactive peptides in their initial form, it is necessary to inhibit these enzymes. Several inhibitors for this purpose have previously been mentioned; however, there has not been any reliable comparison of their efficiency so far. Here, we studied the efficiency of methanol and hydrochloric and formic acids, as well as phenylmethylsulfonyl fluoride, in the inhibition of nine frog peptides with the known sequence, belonging to five families in the secretion of Pelophylax esculentus. The results demonstrated that methanol had the highest inhibitory efficiency, while phenylmethylsulfonyl fluoride was the least efficient, probably due to its instability in aqueous media. Possible cleavages between certain amino acid residues in the sequence were established for each of the inhibitors. These results may be helpful for future studies on the nature of proteases and on prediction of the possible cleavage sites in novel peptides.
Impact of Disease Severity and Disease-Modifying Therapies on Myostatin Levels in SMA Patients
Authors: Mackels Laurane , Mariot Virginie , Buscemi Laura , Servais Laurent , Dumonceaux Julie
Published: August 2024

Clinical trials with treatments inhibiting myostatin pathways to increase muscle mass are currently ongoing in spinal muscular atrophy. Given evidence of potential myostatin pathway downregulation in Spinal Muscular Atrophy (SMA), restoring sufficient myostatin levels using disease-modifying treatments (DMTs) might arguably be necessary prior to considering myostatin inhibitors as an add-on treatment. This retrospective study assessed pre-treatment myostatin and follistatin levels’ correlation with disease severity and explored their alteration by disease-modifying treatment in SMA. We retrospectively collected clinical characteristics, motor scores, and mysotatin and follistatin levels between 2018 and 2020 in 25 Belgian patients with SMA (SMA1 (n = 13), SMA2 (n = 6), SMA 3 (n = 6)) and treated by nusinersen. Data were collected prior to treatment and after 2, 6, 10, 18, and 30 months of treatment. Myostatin levels correlated with patients’ age, weight, SMA type, and motor function before treatment initiation. After treatment, we observed correlations between myostatin levels and some motor function scores (i.e., MFM32, HFMSE, 6MWT), but no major effect of nusinersen on myostatin or follistatin levels over time. In conclusion, further research is needed to determine if DMTs can impact myostatin and follistatin levels in SMA, and how this could potentially influence patient selection for ongoing myostatin inhibitor trials.