bluuebunny/crossref_metadata_2025 huggingface dataset:
Fast Capture and Efficient Removal of Bloom Algae Based on Improved Dielectrophoresis Process
Authors: Liu Jinxin , Jin Qinghao , Geng Junfeng , Xia Jianxin , Wu Yanhong , Chen Huiying
Published: January 2023

A dielectrophoresis (DEP) method for direct capture and fast removal of Anabaena was established in this work. The factors affecting the removal efficiency of Anabaena were investigated systematically, leading to optimized experimental conditions and improved DEP process equipment. The experimental results showed that our improved DEP method could directly capture Anabaena in eutrophic water with much enhanced removal efficiency of Anabaena from high-concentration algal bloom-eutrophication-simulated solution. The removal rate could increase by more than 20% after applying DEP at 15 V compared with a pure filtration process. Moreover, the removal rate could increase from 38.76% to 80.18% in optimized experimental conditions (the initial concentration of 615 μg/L, a flow rate of 0.168 L/h, an AC voltage of 15 V, and frequency of 100 kHz). Optical microscopic images showed that the structure of the captured algae cells was intact, indicating that the DEP method could avoid the secondary pollution caused by the addition of reagents and the release of phycotoxins, providing a new practical method for emergent treatment of water bloom outbreaks.
Prevalence of Body Area Work-Related Musculoskeletal Disorders among Healthcare Professionals: A Systematic Review
Authors: Jacquier-Bret Julien , Gorce Philippe
Published: January 2023

Healthcare professionals perform daily activities that can lead to musculoskeletal disorders (MSDs). The objective of this review was to summarize these MSDs by body areas in relation to healthcare professions. The underlying question is, worldwide, whether there are areas that are more exposed depending on the occupation or whether there are common areas that are highly exposed to MSDs. This issue has been extended to risk factors and responses to reduce MSDs. The review was conducted according to the PRISMA guidelines between February and May 2022. Google scholar and Science Direct databases were scanned to identify relevant studies. Two authors independently reviewed, critically appraised, and extracted data from these studies. Overall and body area prevalence, risk factors, and responses to MSDs were synthetized by occupational activity. Among the 21,766 records identified, 36 covering six healthcare professions were included. The lower back, neck, shoulder and hand/wrist were the most exposed areas for all healthcare professionals. Surgeons and dentists presented the highest prevalence of lower back (>60%), shoulder and upper extremity (35–55%) MSDs. The highest prevalence of MSDs in the lower limbs was found for nurses (>25%). The main causes reported for all healthcare professionals were maintenance and repetition of awkward postures, and the main responses were to modify these postures. Trends by continent seem to emerge regarding the prevalence of MSDs by healthcare profession. Africa and Europe showed prevalence three times higher than Asia and America for lower back MSDs among physiotherapists. African and Asian nurses presented rates three times higher for elbow MSDs than Oceanians. It becomes necessary to objectively evaluate postures and their level of risk using ergonomic tools, as well as to adapt the work environment to reduce exposure to MSDs with regard to the specificities of each profession.
Contraceptive Use and the Associated Factors among Women of Reproductive Age in Jazan City, Saudi Arabia: A Cross-Sectional Survey
Authors: Mahfouz Mohamed Salih , Elmahdy Mona , Ryani Majed Ahmed , Abdelmola Amani Osman , Kariri Samah Ahmed Ali , Alhazmi Hayat Yahya Ahmad , Almalki Salwa Hussain Mater , Adhabi Ons Mohammed , Ali Hindi Sahar Mohammed , Muqri Nouf Mousa , Towhary Bashayer Abdullah
Published: January 2023

The contraceptive use profile is poorly understood in some Gulf Arabian countries, including Saudi Arabia. The present study aimed to assess the knowledge, attitude, and practices related to contraceptive use among women of childbearing age in Jazan city, Saudi Arabia. An observational, analytical cross-sectional study was conducted among a random sample of 450 women in Jazan city. The data were obtained through personal interviews using a questionnaire that included questions on women’s level of knowledge about contraception methods, their attitudes towards contraception methods, and their practices. Data were analyzed using descriptive and inferential statistics techniques using the SPSS program. The prevalence of ever having used contraceptives was significantly different according to age group, job status, children ever born, and the number of pregnancies (p < 0.05 for all). On the other hand, the prevalence of desire to use the contraceptives in the future was high, at 70.9%, with a 95% CI [66.5–74.9]. It differed significantly according to age group and job status (p < 0.05 for both). The most familiar and usable types of contraceptives were pills (36.3%) and intrauterine devices (24.4%). When asked their reasons for using contraceptives, 53.8% of participants cited child spacing and 21.8% improving child health. Logistic regression revealed that the use of contraception among women was more associated with the 20–34 age group [COR = 7.7, 95% CI = 4.4–13.5, p < 0.001] than the 15–24 age group. Having more than one pregnancy and having more than one child were also associated with increased use of contraceptive methods (p < 0.001 for both). These results indicate there is a high level of awareness about contraceptives, a positive attitude toward them, and good practices for the use of them among Saudi women in Jazan. More efforts are needed to improve women’s awareness for better utilization of the available services.
Use of Cardiac Procedures in People with Diabetes during the COVID Pandemic in Spain: Effects on the In-Hospital Mortality
Authors: Lopez-de-Andres Ana , Jimenez-Garcia Rodrigo , Carabantes-Alarcon David , Hernández-Barrera Valentín , de-Miguel-Yanes José M. , de-Miguel-Diez Javier , Zamorano-Leon Jose J. , del-Barrio Jose L. , Cuadrado-Corrales Natividad
Published: January 2023

We aimed to assess the effect of the COVID-19 pandemic in Spain on people with diabetes undergoing cardiac procedures, such as coronary artery bypass graft (CABG), percutaneous coronary intervention (PCI), open surgical valve replacement (OSVR), and transcatheter valve implantation (TCVI). We compared the year 2019 with the year 2020. We conducted an observational study using data from the Spanish National Hospital Discharge Database from 1 January 2019 to 31 December 2020. In 2020, a total of 21,067 cardiac procedures were performed on people with diabetes compared with 24,675 in the previous year. The use of CABG, PCI, OSVR and TCVI decreased from 2019 to 2020 by 13.9%, 14.8%, 21.4% and 2.9%, respectively. In 2020, patients had a significantly higher mean Charlson Comorbidity Index than in 2019 for all the cardiac procedures analyzed. In-hospital mortality (IHM) was higher (p > 0.05) for all the procedures in the year 2020. Over the entire period, female sex was a significant risk factor for IHM among those who underwent CABG, PCI and OSVR (OR 1.94, 95%CI 1.41–2.66; OR 1.19, 95%CI 1.05–1.35; and OR 1.79, 95%CI 1.38–2.32, respectively). The sensitivity analysis including two more years, 2017 and 2018, confirmed that female patients and comorbidity were risk factors for IHM in patients with diabetes regardless of whether it was during the pandemic era or before. We conclude that the frequency of cardiac procedures among people with diabetes declined in 2020. IHM did not change significantly in the COVID-19 era.
Evaluation of the Optimal Manufacturing Protocols and Therapeutic Properties of Mesenchymal Stem/Stromal Cells Derived from Wharton’s Jelly
Authors: Sypecka Monika , Bzinkowska Aleksandra , Sulejczak Dorota , Dabrowski Filip , Sarnowska Anna
Published: January 2023

Wharton’s jelly (WJ) from the umbilical cord (UC) is a good source of mesenchymal stem/stromal cells (MSCs), which can be isolated and used in therapy. Current knowledge shows that even small changes in the cell environment may result in obtaining a subpopulation of cells with different therapeutic properties. For this reason, the conditions of UC transportation, cell isolation, and cultivation and the banking of cells destined for clinical use should be unified and optimized. In this project, we tried various protocols for cell vs. bioptat isolation, banking, and transport in order to determine the most optimal. The most efficient isolation method of WJ-MSCs was chopping the whole umbilical matrix with a scalpel after vessel and lining membrane removal. The optimal solution for short term cell transportation was a multi-electrolyte fluid without glucose. Considering the use of WJ-MSCs in cell therapies, it was important to investigate the soluble secretome of both WJ bioptats and WJ-MSCs. WJ-MSCs secreted higher levels of cytokines and chemokines than WJ bioptats. WJ-MSCs secreted HGF, CCL2, ICAM-1, BDNF, and VEGF. Since these cells might be used in treating neurodegenerative disorders, we investigated the impact of cerebrospinal fluid (CSF) on WJ-MSCs’ features. In the presence of CSF, the cells expressed consecutive neural markers both at the protein and gene level: nestin, β-III-tubulin, S-100-β, GFAP, and doublecortin. Based on the obtained results, a protocol for manufacturing an advanced-therapy medicinal product was composed.
Current Knowledge on the Pathophysiology of Lean/Normal-Weight Type 2 Diabetes
Authors: Salvatore Teresa , Galiero Raffaele , Caturano Alfredo , Rinaldi Luca , Criscuolo Livio , Di Martino Anna , Albanese Gaetana , Vetrano Erica , Catalini Christian , Sardu Celestino , Docimo Giovanni , Marfella Raffaele , Sasso Ferdinando Carlo
Published: January 2023

Since early times, being overweight and obesity have been associated with impaired glucose metabolism and type 2 diabetes (T2D). Similarly, a less frequent adult-onset diabetes in low body mass index (BMI) people has been known for many decades. This form is mainly found in developing countries, whereby the largest increase in diabetes incidence is expected in coming years. The number of non-obese patients with T2D is also on the rise among non-white ethnic minorities living in high-income Western countries due to growing migratory flows. A great deal of energy has been spent on understanding the mechanisms that bind obesity to T2D. Conversely, the pathophysiologic features and factors driving the risk of T2D development in non-obese people are still much debated. To reduce the global burden of diabetes, we need to understand why not all obese people develop T2D and not all those with T2D are obese. Moreover, through both an effective prevention and the implementation of an individualized clinical management in all people with diabetes, it is hoped that this will help to reduce this global burden. The purpose of this review is to take stock of current knowledge about the pathophysiology of diabetes not associated to obesity and to highlight which aspects are worthy of future studies.
Unbalance between Pyridine Nucleotide Cofactors in The SOD1 Deficient Yeast Saccharomyces cerevisiae Causes Hypersensitivity to Alcohols and Aldehydes
Authors: Kwolek-Mirek Magdalena , Bednarska Sabina , Dubicka-Lisowska Aleksandra , Maslanka Roman , Zadrag-Tecza Renata , Kaszycki Pawel
Published: January 2023

Alcohol and aldehyde dehydrogenases are especially relevant enzymes involved in metabolic and detoxification reactions that occur in living cells. The comparison between the gene expression, protein content, and enzymatic activities of cytosolic alcohol and aldehyde dehydrogenases of the wild-type strain and the Δsod1 mutant lacking superoxide dismutase 1, which is hypersensitive to alcohols and aldehydes, shows that the activity of these enzymes is significantly higher in the Δsod1 mutant, but this is not a mere consequence of differences in the enzymatic protein content nor in the expression levels of genes. The analysis of the NAD(H) and NADP(H) content showed that the higher activity of alcohol and aldehyde dehydrogenases in the Δsod1 mutant could be a result of the increased availability of pyridine nucleotide cofactors. The higher level of NAD+ in the Δsod1 mutant is not related to the higher level of tryptophan; in turn, a higher generation of NADPH is associated with the upregulation of the pentose phosphate pathway. It is concluded that the increased sensitivity of the Δsod1 mutant to alcohols and aldehydes is not only a result of the disorder of redox homeostasis caused by the induction of oxidative stress but also a consequence of the unbalance between pyridine nucleotide cofactors.
Oxidative Stress Response’s Kinetics after 60 Minutes at Different (30% or 100%) Normobaric Hyperoxia Exposures
Authors: Leveque Clément , Mrakic-Sposta Simona , Lafère Pierre , Vezzoli Alessandra , Germonpré Peter , Beer Alexandre , Mievis Stéphane , Virgili Fabio , Lambrechts Kate , Theunissen Sigrid , Guerrero François , Balestra Costantino
Published: January 2023

Oxygen is a powerful trigger for cellular reactions and is used in many pathologies, including oxidative stress. However, the effects of oxygen over time and at different partial pressures remain poorly understood. In this study, the metabolic responses of normobaric oxygen intake for 1 h to mild (30%) and high (100%) inspired fractions were investigated. Fourteen healthy non-smoking subjects (7 males and 7 females; age: 29.9 ± 11.1 years, height: 168.2 ± 9.37 cm; weight: 64.4 ± 12.3 kg; BMI: 22.7 ± 4.1) were randomly assigned in the two groups. Blood samples were taken before the intake at 30 min, 2 h, 8 h, 24 h, and 48 h after the single oxygen exposure. The level of oxidation was evaluated by the rate of reactive oxygen species (ROS) and the levels of isoprostane. Antioxidant reactions were observed by total antioxidant capacity (TAC), superoxide dismutase (SOD), and catalase (CAT). The inflammatory response was measured using interleukin-6 (IL-6), neopterin, creatinine, and urates. Oxidation markers increased from 30 min on to reach a peak at 8 h. From 8 h post intake, the markers of inflammation took over, and more significantly with 100% than with 30%. This study suggests a biphasic response over time characterized by an initial “permissive oxidation” followed by increased inflammation. The antioxidant protection system seems not to be the leading actor in the first place. The kinetics of enzymatic reactions need to be better studied to establish therapeutic, training, or rehabilitation protocols aiming at a more targeted use of oxygen.
Polycystin-1 Is a Crucial Regulator of BIN1 Expression and T-Tubule Remodeling Associated with the Development of Dilated Cardiomyopathy
Authors: Díaz-Vesga Magda C. , Flores-Vergara Raúl , Riquelme Jaime A. , Llancaqueo Marcelo , Sánchez Gina , Vergara Cecilia , Michea Luis , Donoso Paulina , Quest Andrew F. G. , Olmedo Ivonne , Pedrozo Zully
Published: January 2023

Cardiomyopathy is commonly observed in patients with autosomal dominant polycystic kidney disease (ADPKD), even when they have normal renal function and arterial pressure. The role of cardiomyocyte polycystin-1 (PC1) in cardiovascular pathophysiology remains unknown. PC1 is a potential regulator of BIN1 that maintains T-tubule structure, and alterations in BIN1 expression induce cardiac pathologies. We used a cardiomyocyte-specific PC1-silenced (PC1-KO) mouse model to explore the relevance of cardiomyocyte PC1 in the development of heart failure (HF), considering reduced BIN1 expression induced T-tubule remodeling as a potential mechanism. PC1-KO mice exhibited an impairment of cardiac function, as measured by echocardiography, but no signs of HF until 7–9 months of age. Of the PC1-KO mice, 43% died suddenly at 7 months of age, and 100% died after 9 months with dilated cardiomyopathy. Total BIN1 mRNA, protein levels, and its localization in plasma membrane-enriched fractions decreased in PC1-KO mice. Moreover, the BIN1 + 13 isoform decreased while the BIN1 + 13 + 17 isoform was overexpressed in mice without signs of HF. However, BIN1 + 13 + 17 overexpression was not observed in mice with HF. T-tubule remodeling and BIN1 score measured in plasma samples were associated with decreased PC1-BIN1 expression and HF development. Our results show that decreased PC1 expression in cardiomyocytes induces dilated cardiomyopathy associated with diminished BIN1 expression and T-tubule remodeling. In conclusion, positive modulation of BIN1 expression by PC1 suggests a novel pathway that may be relevant to understanding the pathophysiological mechanisms leading to cardiomyopathy in ADPKD patients.
iTRAQ-Based Proteomic Analysis of APP Transgenic Mouse Urine Exosomes
Authors: Zhou Xiaojing , Sheikh Abdullah Md. , Matsumoto Ken-ichi , Mitaki Shingo , Shibly Abu Zaffar , Zhang Yuchi , A Garu , Yano Shozo , Nagai Atsushi
Published: January 2023

Alzheimer’s disease (AD) is a common dementia disease in the elderly. To get a better understanding of the pathophysiology, we performed a proteomic analysis of the urine exosomes (U-exo) in AD model mice (J20). The polymer precipitation method was used to isolate U-exo from the urine of 3-month-old J20 and wild-type (WT) mice. Neuron-derived exosome (N-exo) was isolated from U-exo by immunoprecipitation. iTRAQ-based MALDI TOF MS/MS was used for proteomic analysis. The results showed that compared to WT, the levels of 61 and 92 proteins were increased in the J20 U-exo and N-exo, respectively. Gene ontology enrichment analysis demonstrated that the sphingolipid catabolic process, ceramide catabolic process, membrane lipid catabolic process, Aβ clearance, and Aβ metabolic process were highly enriched in U-exo and N-exo. Among these, Asah1 was shown to be the key protein in lipid metabolism, and clusterin, ApoE, neprilysin, and ACE were related to Aβ metabolism and clearance. Furthermore, protein–protein interaction analysis identified four protein complexes where clusterin and ApoE participated as partner proteins. Thus, J20 U-exo and N-exo contain proteins related to lipid- and Aβ-metabolism in the early stages of AD, providing a new insight into the underlying pathological mechanism of early AD.
Disease-Modifying Effects of Non-Invasive Electroceuticals on β-Amyloid Plaques and Tau Tangles for Alzheimer’s Disease
Authors: Bok Junsoo , Ha Juchan , Ahn Bum Ju , Jang Yongwoo
Published: January 2023

Electroceuticals refer to various forms of electronic neurostimulators used for therapy. Interdisciplinary advances in medical engineering and science have led to the development of the electroceutical approach, which involves therapeutic agents that specifically target neural circuits, to realize precision therapy for Alzheimer’s disease (AD). To date, extensive studies have attempted to elucidate the disease-modifying effects of electroceuticals on areas in the brain of a patient with AD by the use of various physical stimuli, including electric, magnetic, and electromagnetic waves as well as ultrasound. Herein, we review non-invasive stimulatory systems and their effects on β-amyloid plaques and tau tangles, which are pathological molecular markers of AD. Therefore, this review will aid in better understanding the recent technological developments, applicable methods, and therapeutic effects of electronic stimulatory systems, including transcranial direct current stimulation, 40-Hz gamma oscillations, transcranial magnetic stimulation, electromagnetic field stimulation, infrared light stimulation and ionizing radiation therapy, and focused ultrasound for AD.
Efficacy of Eribulin Plus Gemcitabine Combination in L-Sarcomas
Authors: López-Álvarez María , González-Aguilera Cristina , Moura David S. , Sánchez-Bustos Paloma , Mondaza-Hernández José L. , Martín-Ruiz Marta , Renshaw Marta , Ramos Rafael , Castilla Carolina , Blanco-Alcaina Elena , Hindi Nadia , Martín-Broto Javier
Published: January 2023

Although the overall survival of advanced soft-tissue sarcoma (STS) patients has increased in recent years, the median progression-free survival is lower than 5 months, meaning that there is an unmet need in this population. Among second-line treatments for advanced STS, eribulin is an anti-microtubule agent that has been approved for liposarcoma. Here, we tested the combination of eribulin with gemcitabine in preclinical models of L-sarcoma. The effect in cell viability was measured by MTS and clonogenic assay. Cell cycle profiling was studied by flow cytometry, while apoptosis was measured by flow cytometry and Western blotting. The activity of eribulin plus gemcitabine was evaluated in in vivo patient-derived xenograft (PDX) models. In L-sarcoma cell lines, eribulin plus gemcitabine showed to be synergistic, increasing the number of hypodiploid events (increased subG1 population) and the accumulation of DNA damage. In in vivo PDX models of L-sarcomas, eribulin combined with gemcitabine was a viable scheme, delaying tumour growth after one cycle of treatment, being more effective in leiomyosarcoma. The combination of eribulin and gemcitabine was synergistic in L-sarcoma cultures and it showed to be active in in vivo studies. This combination deserves further exploration in the clinical context.
Role and Dysregulation of miRNA in Patients with Parkinson’s Disease
Authors: Salemi Michele , Marchese Giovanna , Lanza Giuseppe , Cosentino Filomena I. I. , Salluzzo Maria Grazia , Schillaci Francesca A. , Ventola Giovanna Maria , Cordella Angela , Ravo Maria , Ferri Raffaele
Published: January 2023

Parkinson’s disease (PD) is a neurodegenerative synucleinopathy that has a not yet fully understood molecular pathomechanism behind it. The role of risk genes regulated by small non-coding RNAs, or microRNAs (miRNAs), has also been highlighted in PD, where they may influence disease progression and comorbidities. In this case-control study, we analyzed miRNAs on peripheral blood mononuclear cells by means of RNA-seq in 30 participants, with the aim of identifying miRNAs differentially expressed in PD compared to age-matched healthy controls. Additionally, we investigated the pathways influenced by differentially expressed miRNAs and assessed whether a specific pathway could potentially be associated with PD susceptibility (enrichment analyses performed using the Ingenuity Pathway Analysis tools). Overall, considering that the upregulation of miRNAs might be related with the downregulation of their messenger RNA targets, and vice versa, we found several putative targets of dysregulated miRNAs (i.e., upregulated: hsa-miR-1275, hsa-miR-23a-5p, hsa-miR-432-5p, hsa-miR-4433b-3p, and hsa-miR-4443; downregulated: hsa-miR-142-5p, hsa-miR-143-3p, hsa-miR-374a-3p, hsa-miR-542-3p, and hsa-miR-99a-5p). An inverse connection between cancer and neurodegeneration, called “inverse comorbidity”, has also been noted, showing that some genes or miRNAs may be expressed oppositely in neurodegenerative disorders and in some cancers. Therefore, it may be reasonable to consider these miRNAs as potential diagnostic markers and outcome measures.
Yeast Lipid Produced through Glycerol Conversions and Its Use for Enzymatic Synthesis of Amino Acid-Based Biosurfactants
Authors: Karayannis Dimitris , Papanikolaou Seraphim , Vatistas Christos , Paris Cédric , Chevalot Isabelle
Published: January 2023

The aim of the present work was to obtain microbial lipids (single-cell oils and SCOs) from oleaginous yeast cultivated on biodiesel-derived glycerol and subsequently proceed to the enzymatic synthesis of high-value biosurfactant-type molecules in an aqueous medium, with SCOs implicated as acyl donors (ADs). Indeed, the initial screening of five non-conventional oleaginous yeasts revealed that the most important lipid producer was the microorganism Cryptococcus curvatus ATCC 20509. SCO production was optimised according to the nature of the nitrogen source and the initial concentration of glycerol (Glyc0) employed in the medium. Lipids up to 50% w/w in dry cell weight (DCW) (SCOmax = 6.1 g/L) occurred at Glyc0 ≈ 70 g/L (C/N ≈ 80 moles/moles). Thereafter, lipids were recovered and were subsequently used as ADs in the N-acylation reaction catalysed by aminoacylases produced from Streptomyces ambofaciens ATCC 23877 under aqueous conditions, while Candida antarctica lipase B (CALB) was used as a reference enzyme. Aminoacylases revealed excellent activity towards the synthesis of acyl-lysine only when free fatty acids (FAs) were used as the AD, and the rare regioselectivity in the α-amino group, which has a great impact on the preservation of the functional side chains of any amino acids or peptides. Aminoacylases presented higher α-oleoyl-lysine productivity and final titer (8.3 g/L) with hydrolysed SCO than with hydrolysed vegetable oil. The substrate specificity of both enzymes towards the three main FAs found in SCO was studied, and a new parameter was defined, viz., Specificity factor (Sf), which expresses the relative substrate specificity of an enzyme towards a FA present in a FA mixture. The Sf value of aminoacylases was the highest with palmitic acid in all cases tested, ranging from 2.0 to 3.0, while that of CALB was with linoleic acid (0.9–1.5). To the best of our knowledge, this is the first time that a microbial oil has been successfully used as AD for biosurfactant synthesis. This bio-refinery approach illustrates the concept of a state-of-the-art combination of enzyme and microbial technology to produce high-value biosurfactants through environmentally friendly and economically sound processes.
Employing Engineered Enolase Promoter for Efficient Expression of Thermomyces lanuginosus Lipase in Yarrowia lipolytica via a Self-Excisable Vector
Authors: Jiao Liangcheng , Li Wenjuan , Li Yunchong , Zhou Qinghua , Zhu Mengqin , Zhao Guowei , Zhang Houjin , Yan Yunjun
Published: January 2023

Yarrowia lipolytica is progressively being employed as a workhouse for recombinant protein expression. Here, we expanded the molecular toolbox by engineering the enolase promoter (pENO) and developed a new self-excisable vector, and based on this, a combined strategy was employed to enhance the expression of Thermomyces lanuginosus lipase (TLL) in Y. lipolytica. The strength of 11 truncated enolase promoters of different length was first identified using eGFP as a reporter. Seven of the truncated promoters were selected to examine their ability for driving TLL expression. Then, a series of enolase promoters with higher activities were developed by upstream fusing of different copies of UAS1B, and the recombinant strain Po1f/hp16e100-tll harboring the optimal promoter hp16e100 obtained a TLL activity of 447 U/mL. Additionally, a new self-excisable vector was developed based on a Cre/loxP recombination system, which achieved efficient markerless integration in Y. lipolytica. Subsequently, strains harboring one to four copies of the tll gene were constructed using this tool, with the three-copy strain Po1f/3tll showing the highest activity of 579 U/mL. The activity of Po1f/3tll was then increased to 720 U/mL by optimizing the shaking flask fermentation parameters. Moreover, the folding-related proteins Hac1, Pdi, and Kar2 were employed to further enhance TLL expression, and the TLL activity of the optimal recombinant strain Po1f/3tll-hac1-pdi-kar2 reached 1197 U/mL. By using this combined strategy, TLL activity was enhanced by approximately 39.9-fold compared to the initial strain. Thus, the new vector and the combined strategy could be a useful tool to engineer Y. lipolytica for high-level expression of heterologous protein.
Safety Evaluation and Anti-Inflammatory Efficacy of Lacticaseibacillus paracasei PS23
Authors: Li Chin-Hao , Chen Tai-Ying , Wu Chien-Chen , Cheng Shih-Hsuan , Chang Min-Yu , Cheng Wei-Hong , Chiu Shih-Hau , Chen Chien-Chi , Tsai Ying-Chieh , Yang Deng-Jye , Kang Jaw-Jou , Liao Po-Lin
Published: January 2023

Lacticaseibacillus paracasei strain PS23 (PS23) exhibits some probiotic properties. In this study, a genomic analysis of PS23 revealed no genes related to virulence or antibiotic resistance. Moreover, ornithine decarboxylase activity was not detected in vitro. In addition, PS23 was sensitive to the tested antibiotics. Genotoxicity tests for PS23 including the Ames test and chromosomal aberrations in vitro using Chinese hamster ovary cells and micronuclei in immature erythrocytes of ICR mice were all negative. Moreover, following a 28-day study involving repeated oral dose toxicity tests (40, 400, and 4000 mg/kg equal 1.28 × 1010, 1.28 × 1011, and 1.28 × 1012 CFU/kg body weight, respectively) using an ICR mouse model, no adverse effects were observed from any doses. In addition, supplementation with live or heat-killed PS23 ameliorates DSS-induced colonic inflammation in mice. Our findings suggest that PS23 is safe and has anti-inflammatory effects and may therefore have therapeutic implications.
Improvement of a Surfactant Blend for Enhanced Oil Recovery in Carbonate Reservoirs by Means of an Ionic Liquid
Authors: Tafur Nestor , Muñuzuri Alberto P. , Soto Ana
Published: January 2023

The promising experimental performance of surfactant blends encourages their use in recovering the large quantity of crude oil still remaining in carbonate reservoirs. Phase behavior studies were carried out in this work to propose a blend for practical application. To that aim, the surfactants dioctyl sulfosuccinate sodium (AOT) and polyoxyethylene(8) octyl ether carboxylic acid (Akypo LF2) were mixed. A formulation consisting of 1 wt% of AOT50wt%/LF250wt% blend in synthetic sea water (SSW) led to a low value of interfacial tension with crude oil of 1.50·10−2 mN/m, and 0.42 mg/grock of dynamic adsorption. A moderate additional oil recovery (7.3% of the original oil in place) was achieved in a core flooding test. To improve this performance, the surface-active ionic liquid 1-dodecyl-3-methylimidazolium bromide ([C12mim]Br) was added to the system. The electrostatic interactions between the oppositely charged surfactants (AOT and [C12mim]Br) led to a higher surface activity. Thus, a formulation consisting of 0.8 wt% of AOT20.7wt%/[C12mim]Br25.3wt%/LF254wt% in SSW reduced the interfacial tension and surfactant adsorption achieved with the binary blend to 1.14 × 10−2 mN/m and 0.21 mg/grock, respectively. The additional oil recovery achieved with the blend containing the ionic liquid was 11.5% of the original oil in place, significantly improving the efficiency of the binary blend.
LUHMES Cells: Phenotype Refinement and Development of an MPP+-Based Test System for Screening Antiparkinsonian Drugs
Authors: Beliakov Sergei V. , Blokhin Victor , Surkov Sergey A. , Ugrumov Michael V.
Published: January 2023

The low effectiveness of symptomatic pharmacotherapy for Parkinson’s disease (PD), which compensates for dopamine (DA) deficiency under degeneration of nigrostriatal dopaminergic (DAergic) neurons, could apparently be improved with neuroprotective therapy, which slows down neurodegeneration and PD progression. For this, it is necessary to have a DAergic cell line for the development of a PD model to screen neuroprotectors. We used immortalized human embryonic mesencephalon LUHMES cells (LCs) differentiated into DAergic neurons. The aim of this study was to characterize the phenotype of differentiated LCs and develop an 1-methyl-4-phenylpyridinium iodide (MPP+)-based test system for screening neuroprotectors. Using polymerase chain reaction (PCR) and immunocytochemistry, it has been shown that all differentiated LCs express genes and synthesize proteins characteristic of all neurons (microtubule-associated protein 2, bIII-tubulin, synaptotagmin 1) and specifically of DAergic neurons (tyrosine hydroxylase, aromatic L-amino acid decarboxylase, DA transporter, vesicular monoamine transporter 2). Furthermore, LCs are able to produce a small amount of DA, but under special conditions. To assess the mechanisms of neurodegeneration and neuroplasticity under the influence of toxins and antiparkinsonian drugs, including neuroprotectors, we have developed an LCs-based MPP+ PD model and proposed an original panel of markers for testing functional and structural cell disorders.
Anammox Bacterial S-Adenosyl-l-Methionine Dependent Methyltransferase Crystal Structure and Its Interaction with Acyl Carrier Proteins
Authors: Uegaki Tesshin , Takei Taisei , Yamaguchi Shuhei , Fujiyama Keisuke , Sato Yusuke , Hino Tomoya , Nagano Shingo
Published: January 2023

Ladderane lipids (found in the membranes of anaerobic ammonium-oxidizing [anammox] bacteria) have unique ladder-like hydrophobic groups, and their highly strained exotic structure has attracted the attention of scientists. Although enzymes encoded in type II fatty acid biosynthesis (FASII) gene clusters in anammox bacteria, such as S-adenosyl-l-methionine (SAM)-dependent enzymes, have been proposed to construct a ladder-like structure using a substrate connected to acyl carrier protein from anammox bacteria (AmxACP), no experimental evidence to support this hypothesis was reported to date. Here, we report the crystal structure of a SAM-dependent methyltransferase from anammox bacteria (AmxMT1) that has a substrate and active site pocket between a class I SAM methyltransferase-like core domain and an additional α-helix inserted into the core domain. Structural comparisons with homologous SAM-dependent C-methyltransferases in polyketide synthase, AmxACP pull-down assays, AmxACP/AmxMT1 complex structure predictions by AlphaFold, and a substrate docking simulation suggested that a small compound connected to AmxACP could be inserted into the pocket of AmxMT1, and then the enzyme transfers a methyl group from SAM to the substrate to produce branched lipids. Although the enzymes responsible for constructing the ladder-like structure remain unknown, our study, for the first time, supports the hypothesis that biosynthetic intermediates connected to AmxACP are processed by SAM-dependent enzymes, which are not typically involved in the FASII system, to produce the ladder-like structure of ladderane lipids in anammox bacteria.
Particle Size Modulates Silver Nanoparticle Toxicity during Embryogenesis of Urchins Arbacia lixula and Paracentrotus lividus
Authors: Burić Petra , Čarapar Ivana , Pavičić-Hamer Dijana , Kovačić Ines , Jurković Lara , Dutour Sikirić Maja , Domazet Jurašin Darija , Mikac Nevenka , Bačić Niko , Lyons Daniel Mark
Published: January 2023

Silver nanoparticles represent a threat to biota and have been shown to cause harm through a number of mechanisms, using a wide range of bioassay endpoints. While nanoparticle concentration has been primarily considered, comparison of studies that have used differently sized nanoparticles indicate that nanoparticle diameter may be an important factor that impacts negative outcomes. In considering this, the aim of the present study was to determine if different sizes of silver nanoparticles (AgNPs; 10, 20, 40, 60 and 100 nm) give rise to similar effects during embryogenesis of Mediterranean sea urchins Arbacia lixula and Paracentrotus lividus, or if nanoparticle size is a parameter that can modulate embryotoxicity and spermiotoxicity in these species. Fertilised embryos were exposed to a range of AgNP concentrations (1–1000 µg L−1) and after 48 h larvae were scored. Embryos exposed to 1 and 10 µg L−1 AgNPs (for all tested sizes) showed no negative effect in both sea urchins. The smaller AgNPs (size 10 and 20 nm) caused a decrease in the percentage of normally developed A. lixula larvae at concentrations ≥50 µg L−1 (EC50: 49 and 75 μg L−1, respectively) and at ≥100 µg L−1 (EC50: 67 and 91 μg L−1, respectively) for P. lividus. AgNPs of 40 nm diameter was less harmful in both species ((EC50: 322 and 486 μg L−1, for P. lividus and A. lixula, respectively)). The largest AgNPs (60 and 100 nm) showed a dose-dependent response, with little effect at lower concentrations, while more than 50% of larvae were developmentally delayed at the highest tested concentrations of 500 and 1000 µg L−1 (EC50(100 nm); 662 and 529 μg L−1, for P. lividus and A. lixula, respectively. While AgNPs showed no effect on the fertilisation success of treated sperm, an increase in offspring developmental defects and arrested development was observed in A. lixula larvae for 10 nm AgNPs at concentrations ≥50 μg L−1, and for 20 and 40 nm AgNPs at concentrations >100 μg L−1. Overall, toxicity was mostly ascribed to more rapid oxidative dissolution of smaller nanoparticles, although, in cases, Ag+ ion concentrations alone could not explain high toxicity, indicating a nanoparticle-size effect.
α4-α5 Helices on Surface of KRAS Can Accommodate Small Compounds That Increase KRAS Signaling While Inducing CRC Cell Death
Authors: Abuasaker Baraa , Garrido Eduardo , Vilaplana Marta , Gómez-Zepeda Jesús Daniel , Brun Sonia , Garcia-Cajide Marta , Mauvezin Caroline , Jaumot Montserrat , Pujol Maria Dolors , Rubio-Martínez Jaime , Agell Neus
Published: January 2023

KRAS is the most frequently mutated oncogene associated with the genesis and progress of pancreatic, lung and colorectal (CRC) tumors. KRAS has always been considered as a therapeutic target in cancer but until now only two compounds that inhibit one specific KRAS mutation have been approved for clinical use. In this work, by molecular dynamics and a docking process, we describe a new compound (P14B) that stably binds to a druggable pocket near the α4-α5 helices of the allosteric domain of KRAS. This region had previously been identified as the binding site for calmodulin (CaM). Using surface plasmon resonance and pulldown analyses, we prove that P14B binds directly to oncogenic KRAS thus competing with CaM. Interestingly, P14B favors oncogenic KRAS interaction with BRAF and phosphorylated C-RAF, and increases downstream Ras signaling in CRC cells expressing oncogenic KRAS. The viability of these cells, but not that of the normal cells, is impaired by P14B treatment. These data support the significance of the α4-α5 helices region of KRAS in the regulation of oncogenic KRAS signaling, and demonstrate that drugs interacting with this site may destine CRC cells to death by increasing oncogenic KRAS downstream signaling.
Dexmedetomidine Attenuates Methotrexate-Induced Neurotoxicity and Memory Deficits in Rats through Improving Hippocampal Neurogenesis: The Role of miR-15a/ROCK-1/ERK1/2/CREB/BDNF Pathway Modulation
Authors: Taha Mohamed , Eldemerdash Omar Mohsen , Elshaffei Ismail Mohamed , Yousef Einas Mohamed , Senousy Mahmoud A.
Published: January 2023

Methotrexate (MTX) is a widely used neurotoxic drug with broad antineoplastic and immunosuppressant spectra. However, the exact molecular mechanisms by which MTX inhibits hippocampal neurogenesis are yet unclear. Dexmedetomidine (Dex), an α2-adrenergic receptor agonist, has recently shown neuroprotective effects; however, its full mechanism is unexplored. This study investigated the potential of Dex to mitigate MTX-induced neurotoxicity and memory impairment in rats and the possible role of the miR-15a/ROCK-1/ERK1/2/CREB/BDNF pathway. Notably, no former studies have linked this pathway to MTX-induced neurotoxicity. Male Sprague Dawley rats were placed into four groups. Group 1 received saline i.p. daily and i.v. on days 8 and 15. Group 2 received Dex at 10 μg/kg/day i.p. for 30 days. Group 3 received MTX at 75 mg/kg i.v. on days 8 and 15, followed by four i.p. doses of leucovorin at 6 mg/kg after 18 h and 3 mg/kg after 26, 42, and 50 h. Group 4 received MTX and leucovorin as in group 3 and Dex daily dosages as in group 2. Bioinformatic analysis identified the association of miR-15a with ROCK-1/ERK1/2/CREB/BDNF and neurogenesis. MTX lowered hippocampal doublecortin and Ki-67, two markers of neurogenesis. This was associated with the downregulation of miR-15a, upregulation of its target ROCK-1, and reduction in the downstream ERK1/2/CREB/BDNF pathway, along with disturbed hippocampal redox state. Novel object recognition and Morris water maze tests demonstrated the MTX-induced memory deficiencies. Dex co-treatment reversed the MTX-induced behavioral, biochemical, and histological alterations in the rats. These neuroprotective actions could be partly mediated through modulating the miR-15a/ROCK-1/ERK1/2/CREB/BDNF pathway, which enhances hippocampal neurogenesis.
Blood Plasma Proteome: A Meta-Analysis of the Results of Protein Quantification in Human Blood by Targeted Mass Spectrometry
Authors: Kliuchnikova Anna A. , Novikova Svetlana E. , Ilgisonis Ekaterina V. , Kiseleva Olga I. , Poverennaya Ekaterina V. , Zgoda Victor G. , Moshkovskii Sergei A. , Poroikov Vladimir V. , Lisitsa Andrey V. , Archakov Alexander I. , Ponomarenko Elena A.
Published: January 2023

A meta-analysis of the results of targeted quantitative screening of human blood plasma was performed to generate a reference standard kit that can be used for health analytics. The panel included 53 of the 296 proteins that form a “stable” part of the proteome of a healthy individual; these proteins were found in at least 70% of samples and were characterized by an interindividual coefficient of variation <40%. The concentration range of the selected proteins was 10−10–10−3 M and enrichment analysis revealed their association with rare familial diseases. The concentration of ceruloplasmin was reduced by approximately three orders of magnitude in patients with neurological disorders compared to healthy volunteers, and those of gelsolin isoform 1 and complement factor H were abruptly reduced in patients with lung adenocarcinoma. Absolute quantitative data of the individual proteome of a healthy and diseased individual can be used as the basis for personalized medicine and health monitoring. Storage over time allows us to identify individual biomarkers in the molecular landscape and prevent pathological conditions.
Apoe4 and Alzheimer’s Disease Pathogenesis—Mitochondrial Deregulation and Targeted Therapeutic Strategies
Authors: Pires Mariana , Rego Ana Cristina
Published: January 2023

APOE ε4 allele (ApoE4) is the primary genetic risk factor for sporadic Alzheimer’s disease (AD), expressed in 40–65% of all AD patients. ApoE4 has been associated to many pathological processes possibly linked to cognitive impairment, such as amyloid-β (Aβ) and tau pathologies. However, the exact mechanism underlying ApoE4 impact on AD progression is unclear, while no effective therapies are available for this highly debilitating neurodegenerative disorder. This review describes the current knowledge of ApoE4 interaction with mitochondria, causing mitochondrial dysfunction and neurotoxicity, associated with increased mitochondrial Ca2+ and reactive oxygen species (ROS) levels, and it effects on mitochondrial dynamics, namely fusion and fission, and mitophagy. Moreover, ApoE4 translocates to the nucleus, regulating the expression of genes involved in aging, Aβ production, inflammation and apoptosis, potentially linked to AD pathogenesis. Thus, novel therapeutical targets can be envisaged to counteract the effects induced by ApoE4 in AD brain.
The Establishment of a Genetic Transformation System and the Acquisition of Transgenic Plants of Oriental Hybrid Lily (Lilium L.)
Authors: Chen Yue , Hou Xinru , Zheng Yupin , Lyu Yingmin
Published: January 2023

Lily (Lilium spp.) has elegant flowers and beautiful colors, which makes it popular among people. However, the poor stress resistance and self-propagation ability of lily limit its application in landscaping to a great extent. In addition, transgenic technology is an important means to improve plant characteristics, but the lack of a stable and efficient genetic transformation system is still an important factor restricting the development of lily transgenic technology. Therefore, this study established a good lily regeneration system by screening different explants and plant growth regulators of different concentrations. Then, the genetic transformation system of lily was optimized by screening the critical concentration of antibiotics, the concentration of bacterial solution, and the infection time. Finally, the homologous lily cold resistance gene LlNAC2 and bulblet generation gene LaKNOX1 were successfully transferred to ‘Siberia’ and ‘Sorbonne’ to obtain lily transgenic lines. The results showed that when the stem axis was used as explant in ‘Siberia’, the induction rate was as high as 87%. The induction rate of ‘Sorbonne’ was as high as 91.7% when the filaments were used as explants. At the same time, in the optimized genetic transformation system, the transformation rate of ‘Siberia’ and ‘Sorbonne’ was up to 60%. In conclusion, this study provides the theoretical basis and technical support for improving the resistance and reproductive ability of Oriental lily and the molecular breeding of lily.
Keeping the Spirit in the Bottle: On Pathological Reduction of Information in Totalitarianism
Authors: Postoutenko Kirill
Published: January 2023

This article begins with disputing the teleologically charged notion of unstoppable information growth, pointing at the alternation of informational contraction and expansion in open dynamic systems. Narrowing the focus, it turns to the 20th century totalitarian systems as particularly paradoxical informational environments: Being less capable of processing information than their democratic counterparts and therefore more vulnerable to overloads, they are particularly prone to suppressing informational transmission in some areas, codes and media. Dilution and conflation are singled out as the most common ways of lessening the informational value of communication in totalitarian societies. Whereas the first greatly increases the ratios of signs to messages and messages to interactions, causing redundancy and semantic inflation, the second rolls back preexisting functional differentiations (person vs. social role, sender vs. message, message vs. information etc.) within societies and their communicative system. It is argued that both attempts at semantic impoverishment of public communication in totalitarianism lead to the pathological states, failing to reduce the overall amount of information within the systems in question and precipitating the very informational explosions they were designed to prevent.
Synthesis, DFT and X-ray Studies of Trans CuCl2L2 with L Is (E)-(4-Chlorophenyl)-N-(3-phenyl-4H-1,2,4-triazol-4-yl)methanimine
Authors: Hammud Hassan H. , Wehbie Moheddine , Abdul-Ghani Mohamed M. , Gal Zoltan A. , Sheikh Abdul Hamid Malai Haniti , Sheikh Nadeem S.
Published: January 2023

A novel approach was carried to prepare trans-CuCl2L2 complex with the ligand L, (E)-(4-chlorophenyl)-N-(3-phenyl-4H-1,2,4-triazol-4-yl)methanimine which was formed in situ during the reaction of CuCl2 with 4-(4-chlorobenzylideneamino)-5-phenyl-2H-1,2,4-triazole-3(4H)-thione. The synthesized compounds were characterized by applying various spectroscopic techniques. The crystal structure of the complex was unambiguously determined using X-ray analysis indicating square planar geometry. Intermolecular H-bonds govern the supramolecular structure of the copper complex. Aromatic rings are stacked in an offset packing due to occurrence of π–π interactions. The structure is further corroborated with a detailed computational investigation. A thione–thiol tautomerism for the triazole compound was also studied. The Schiff base 1,2,4-triazole copper chloride complex is expected to have high anticancer activity.