bluuebunny/crossref_metadata_2025 huggingface dataset:
Study on the Properties and Heavy Metal Solidification Characteristics of Sintered Ceramsites Composed of Magnesite Tailings, Sewage Sludge, and Coal Gangue
Authors: Li Yanlong , Xu Mingyuan , Li Quan , Gai Anjun , Yang Tianhua , Li Rundong
Published: September 2022

With the rapid development of industry, the disposal of industrial solid waste needs to be solved urgently in China. Thus, an effective disposal method should be proposed to recycle these solid wastes in an environmentally friendly and sustainable manner. In this paper, ceramsite was prepared from sewage sludge (SS), magnesite tailings (MTs), and coal gangue (CG). The influence of the material ratio and sintering temperature on the properties of the ceramsite was investigated. The results show that the ceramsite had better properties when the following parameters were used: a ratio of SS: CG: MT of 4.5:4:1.5; a sintering temperature of 1250 °C; a compressive strength of 11.2 MPa (or it can be rounded to 11; our major remark relates to significant figures, and they should be up to 2–3 figures, according to measurement errors); a water absorption of 3.54%; and apparent and bulk densities of 1.19 and 0.81 g/cm3, respectively. The strength was superior to more than twice the 900-density grade prescribed by the Chinese national standard. After sintering, most of the heavy metals in the ceramsite mainly existed in the form of residue state (FD), meaning that they were highly stable. The leaching concentrations of Zn and Ni from the ceramsite were 0.72 and 0.25 mg/L lower than the prescribed regulatory limits (2.0 and 0.1 mg/L). The overall pollution toxicity index (OPTI) was only 240, less than that of raw pellets, indicating that the environmental risk is low. Not only did the ceramsite, prepared from SS, CG, and MT, exhibit excellent chemical properties, but it also proved to be an environmentally safe material. Therefore, it is an effective approach to realize the collaborative treatment of SS, CG, and MT by preparing ceramsite.
Boosting Sustainable Operations with Sustainable Supply Chain Modeling: A Case of Organizational Culture and Normative Commitment
Authors: Lazar Sebastjan , Potočan Vojko , Klimecka-Tatar Dorota , Obrecht Matevz
Published: September 2022

The importance of sustainability in supply chain management is growing worldwide. It is possible to find reasons for this using various phenomena that negatively affect humanity, e.g., climate change, scarce materials, supply disruptions, and complex fossil fuel dependency. Because of that, is extremely important to constantly look for new ways to systematically increase sustainability in enterprises and their logistics and supply chain processes by considering different stakeholders and influential factors. Therefore, this paper explores how different types of organizational culture and normative commitment impact sustainability and each other in business logistics and supply chains and develops a conceptual model to manage this challenge. Gaining new insights is valuable especially for managers to obtain better information on how to improve sustainability not just by integrating green technologies but mainly by changing culture, attitude, and perception in their enterprises. The research is focused on employees from global logistics or related branches in micro, small, medium, and large enterprises with the primary activity mostly related to manufacturing, transport, and storage. The findings are based on the questionnaire which was sent directly to 1576 employees from 528 enterprises. A total of 516 employees from enterprises that are mostly located in 34 countries responded to requests for participation. The results reveal statistically significant positive and negative impacts, e.g., clan culture has a positive statistically significant impact on the sustainable development of supply chains. Most of the connections to the eighth Sustainable Development Goal by the United Nations (decent work and economic growth) were also found, which was the enterprise’s highest priority with a share of 52.99%. A contribution to the theory development is gained using the developed model that considers both positive and negative statistically significant impacts studied.
A Statistical Model of COVID-19 Infection Incidence in the Southern Indian State of Tamil Nadu
Authors: Devi Tanmay , Gopalan Kaushik
Published: September 2022

In this manuscript, we present an analysis of COVID-19 infection incidence in the Indian state of Tamil Nadu. We used seroprevalence survey data along with COVID-19 fatality reports from a six-month period (1 June 2020 to 30 November 2020) to estimate age- and sex-specific COVID-19 infection fatality rates (IFR) for Tamil Nadu. We used these IFRs to estimate new infections occurring daily using the daily COVID-19 fatality reports published by the Government of Tamil Nadu. We found that these infection incidence estimates for the second COVID wave in Tamil Nadu were broadly consistent with the infection estimates from seroprevalence surveys. Further, we propose a composite statistical model that pairs a k-nearest neighbours model with a power-law characterisation for “out-of-range” extrapolation to estimate the COVID-19 infection incidence based on observed cases and test positivity ratio. We found that this model matched closely with the IFR-based infection incidence estimates for the first two COVID-19 waves for both Tamil Nadu as well as the neighbouring state of Karnataka. Finally, we used this statistical model to estimate the infection incidence during the recent “Omicron wave” in Tamil Nadu and Karnataka.
Potential Health Risk to Brazilian Infants by Polybrominated Diphenyl Ethers Exposure via Breast Milk Intake
Authors: Souza Marília , Devóz Paula , Ximenez João , Bocato Mariana , Rocha Bruno , Barbosa Fernando
Published: September 2022

Polybrominated diphenyl ethers (PBDEs) are ubiquitous flame retardants and are environmentally persistent. PBDEs show endocrine disruption, neurotoxicity, and lower birth weight in infants, and their human body burden has become a public health concern. The infants’ exposure begins in the prenatal period and continues via breast milk ingestion, although, little is known about the factors that may influence this exposure. In this study, PBDE levels in Brazilian breast milk were assessed in 200 lactating women. The risk assessment of infants’ exposure to PBDE was performed through the estimated daily intake (EDI) calculation. The geometric mean (GM) of ∑PBDEs levels was 2.33 (0.14–6.05) ng/g wet weight. At least one PBDE congener was detected in the samples, and the 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) showed a 100% of detection rate (GM of 1.05 ng/g). Location of residence, maternal level education, monthly salary, and race were positively associated with PBDE levels (p < 0.05). The EDI of BDE-47 was higher in Belo Horizonte (8.29 ng/kg/day) than in Viçosa (6.36 ng/kg/day), as well as for the ∑PBDEs (19.77 versus 12.78 ng/kg/day) (p < 0.05). Taking the high detection rate of PBDEs in breast milk and their toxicity, continuous studies on infant exposure, fetal growth, and child neurodevelopment are requested.
Attitudes, Perceptions and Practices of Influenza Vaccination in the Adult Population: Results of a Cross-Sectional Survey in Spain
Authors: Prada-García Camino , Fernández-Espinilla Virginia , Hernán-García Cristina , Sanz-Muñoz Iván , Martínez-Olmos José , Eiros Jose M. , Castrodeza-Sanz Javier
Published: September 2022

In Spain, the 2021/22 influenza season overlapped with the sixth wave of the 2019 coronavirus disease pandemic (COVID-19). Influenza is a major public health problem associated with high morbidity and mortality. The objectives of this study were to determine the knowledge, perceptions and practices of influenza vaccination in the Spanish population, coinciding with the COVID-19 pandemic, with special attention paid to people over 65 years of age. A cross-sectional study was carried out by conducting 2211 telephone interviews. It was observed that 81.6% of people ≥ 65 years were vaccinated annually or with some frequency compared to 35.5% of those under 65 years (p < 0.001). Fifty percent of Spaniards showed an intention to be vaccinated in the 2021/22 campaign, during the SARS-CoV2 pandemic. In the case of people ≥ 65 years old, this figure was 83% compared to 42% of those under 65 years old (p < 0.001). Significant predictors of intention to be vaccinated were age of 65 years or older (OR 1.8, 95% CI 1.3–2.5), female sex (OR 1.9, 95% CI 1.5–2.4), belonging to risk groups (OR 2.2, 95% CI 1.6–3.1) and having been previously vaccinated (OR 29.7, 95% CI 22.5–39.2). The main reasons for deciding to be vaccinated were the need to be protected against the virus and to be vaccinated annually. On the other hand, lack of recommendation and considering the influenza vaccine as not necessary were the main reasons for not getting vaccinated. In addition, health personnel stood out as the main source of information (32.9%) compared to traditional media (26.9%) and public administration (12.3%). This study aimed to assess and analyse the factors influencing willingness to receive influenza vaccines in the COVID-19 era among Spanish adults, as well as the main information channels and strategies to encourage vaccination.
Person-Centered Practice in Hospitalized Older Adults with Chronic Illness: Clinical Study Protocol
Authors: Vareta Diana Alves , Ventura Filipa , Família Carlos , Oliveira Célia
Published: September 2022

The aging trend in the population, the high rate of hospitalization, the affliction by multiple chronic illnesses, and the increased vulnerability of older people when hospitalized undoubtedly require a person-centered approach to healthcare—an approach that values a person’s participation in the healthcare relationship, supports shared decision making and mutual understanding, and respects a person’s values, preferences, and beliefs. However, despite widespread recognition that the adoption of such a clinical practice paradigm is paramount, its implementation and development are still challenging for various health systems and professionals worldwide. The implementation strategy for such a healthcare paradigm must be based on each country’s health system organization and practice contexts, as well as the professionals involved. The present work aims to provide guidelines for the understanding of the state of development of person-centered practice in the daily care of hospitalized older adults with chronic illnesses within the internal medicine department of a secondary hospital in an urban area of Portugal. We focus on the characterization of (i) the perceptions of a multidisciplinary team working at an inpatient hospital department of person-centered practice, (ii) the perceptions of hospitalized older adults with chronic illnesses about person-centered practice, (iii) the work culture of an inpatient hospital department with a high prevalence of older adults with chronic illnesses, (iv) the Person-Centred Practice Framework at the organizational and structural levels of the healthcare system, and (v) the elements that influence the implementation of person-centered practice at the individual, organizational, and structural levels in this specific hospital context. To this end, a mixed-methods analysis with a convergent design was planned to use questionnaire instruments to collect data in parallel and independently from distinct samples of health professionals and older inpatient adults within this department. Furthermore, health policies and strategic plans will be analyzed to identify and evaluate references and guidelines for the practice of person-centered care. Studying the dimensions of clinical practice in this specific healthcare context following the Person-Centred Practice Framework can allow us to understand the extent of its development in terms of prerequisites, care environment, care processes, and the macro-context of the healthcare system. Therefore, it is possible to identify and characterize the dimensions achieved and those that need to be improved and, thus, establish a starting point for the definition of new strategies to advance practice towards person-centeredness and monitor changes in healthcare practice.
Hair Toxic Trace Elements of Residents across the Caspian Oil and Gas Region of Kazakhstan: Cross-Sectional Study
Authors: Batyrova Gulnara , Tlegenova Zhenisgul , Kononets Victoria , Umarova Gulmira , Kudabayeva Khatimya , Bazargaliyev Yerlan , Amanzholkyzy Ainur , Umarov Yeskendir
Published: September 2022

This study aimed to assess the relationship between the content of toxic trace elements, such as aluminum (Al), arsenic (As), beryllium (Be), cadmium (Cd), mercury (Hg), and lead (Pb), in the hair of the adult population of western Kazakhstan and the distance of their residence from oil and gas fields. The cross-sectional study included 850 adults aged 18–60 years. Inductively coupled plasma mass spectrometry was used to measure the level of Al, As, Be, Cd, Hg, and Pb in hair. The relationship between the concentration of toxic trace elements in the hair and the distance from oil and gas fields was assessed in three groups (<16 km, 16–110 km, and >110 km), using multiple linear regression analysis. The highest concentration of Hg = 0.338 μg/g was determined in the group living near oil and gas fields (0–16 km), whereas the lowest concentration of Al = 3.127 μg/g and As = 0.028 μg/g was determined in participants living at a long distance (more than 110 km) (p < 0.001). The concentration of Al (−0.126 (CI: −0.174; −0.077)), Hg (−0.065 (CI: −0.129; −0.001)), and Pb (0.111 (CI: 0.045; 0.177)) is associated with the distance to oil and gas fields. The obtained data indicate a change in the toxic trace element content in the hair of residents in the Caspian region of western Kazakhstan, a change that is most pronounced in residents living in the zone of oil and gas pollution. The distance to the oil and gas fields affects the content of toxic elements in scalp hair. In particular, the concentration of Al and Hg is associated with a decrease in the distance to oil and gas fields, while the concentration of Pb is associated with an increase in the distance to these fields. The lowest content of Al and As was determined in the hair of study participants living in the most remote areas (more than 110 km from oil and gas fields). Our results demonstrate the need for the biomonitoring of toxic elements to determine long-term temporal trends in the impact of chemicals on public health in western Kazakhstan.
Molecular Pathways in Pulmonary Arterial Hypertension
Authors: Shah Aangi J. , Vorla Mounica , Kalra Dinesh K.
Published: September 2022

Pulmonary arterial hypertension is a multifactorial, chronic disease process that leads to pulmonary arterial endothelial dysfunction and smooth muscular hypertrophy, resulting in impaired pliability and hemodynamics of the pulmonary vascular system, and consequent right ventricular dysfunction. Existing treatments target limited pathways with only modest improvement in disease morbidity, and little or no improvement in mortality. Ongoing research has focused on the molecular basis of pulmonary arterial hypertension and is going to be important in the discovery of new treatments and genetic pathways involved. This review focuses on the molecular pathogenesis of pulmonary arterial hypertension.
Lipopolysaccharide from Porphyromonas gingivalis, but Not from Porphyromonas endodontalis, Induces Macrophage M1 Profile
Authors: Veloso Pablo , Fernández Alejandra , Astorga Jessica , González-Quintanilla David , Castro Alfredo , Escobar Alejandro , Hoare Anilei , Hernández Marcela
Published: September 2022

Apical Lesions of Endodontic Origin (ALEO) are initiated by polymicrobial endodontic canal infection. Porphyromonas gingivalis (Pg) and Porphyromonas endodontalis (Pe) lipopolysaccharides (LPS) can induce a pro-inflammatory macrophage response through their recognition by TLR2 and TLR4. However, polarization responses induced by Pg and/or Pe LPS in macrophages are not fully understood. We aimed to characterize the polarization profiles of macrophages differentiated from THP-1 cells following Pg and/or Pe LPS stimulation from reference strain and clinical isolates. A modified LPS purification protocol was implemented and the electrophoretic LPS profiles were characterized. THP-1 human monocytes differentiated to macrophages were stimulated with Pg and Pe LPS. Polarization profiles were characterized through cell surface markers and secreted cytokines levels after 24 h of stimulation. TLR2 and TLR4 cell surfaces and transcriptional levels were determined after 24 or 2 h of LPS stimulation, respectively. LPS from Pg induced a predominant M1 profile in macrophages evidenced by changes in the expression of the surface marker CD64 and pro-inflammatory cytokine profiles, TNF-α, IL-1β, IL-6, and IL-12. Pe LPS was unable to induce a significant response. TLR2 and TLR4 expressions were neither modified by Pg or Pe LPS. Pg LPS, but not Pe LPS, induced a macrophage M1 Profile.
Functional Characterization of Two RNA Methyltransferase Genes METTL3 and METTL14 Uncovers the Roles of m6A in Mediating Adaptation of Plutella xylostella to Host Plants
Authors: Wang Bei-Bei , Lai Ying-Fang , Li Fei-Fei , Jiao Lu , Qiao Qing-Xuan , Li Shan-Yu , Xiang Xiu-Juan , Liao Huang , You Min-Sheng , He Wei-Yi
Published: September 2022

N6-methyladenosine (m6A) is one of the major epigenetic modifications in eukaryotes. Although increasing functions of m6A have been identified in insects, its role in Plutella xylostella L. for host plant adaptation remains unclear. In the current study, we show that the m6A content of P. xylostella was relatively low in different developmental stages and tissues, with no significant differences. Two RNA methyltransferase genes, PxMETTL3 (methyltransferase-like 3) and PxMETTL14 (methyltransferase-like 14), were identified and characterized. PxMETTL3 could be transcribed into two transcripts, and PxMETTL14 had only one transcript; both of these genes were highly expressed in egg and adult stages and reproductive tissues. The CRISPR/Cas9-mediated knockout of PxMETTL3 (ΔPxMETTL3-2) or PxMETTL14 (ΔPxMETTL14-14) confirmed their function in m6A installation into RNA. Furthermore, upon transfer from an artificial diet to the host plant, the mutant strains were affected in terms of larval and pupal weight or adult emergence rate, while the wildtype (WT) strain did not exhibit any difference. In addition, the fecundity and egg hatching rate of the WT strain decreased significantly, whereas only the ΔPxMETTL14-14 mutant strain displayed significantly decreased fecundity. There seemed to be a tradeoff between the stress adaptation and reproduction in P. xylostella mediated by m6A modification. During host transfer, the expression of PxMETTL14 was consistent with the change in m6A content, which implied that PxMETTL14 could respond to host plant defense effectively, and may regulate m6A content. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the differentially expressed transcripts with changes in m6A levels revealed that the potential functions of m6A-related genes may be involved in steroid biosynthesis for larval performance and metabolic pathways for adult reproduction. Overall, our work reveals an epigenetic regulation mechanism for the rapid adaptation of P. xylostella to variations in the host environment.
Biological Features and In Planta Transcriptomic Analyses of a Microviridae Phage (CLasMV1) in “Candidatus Liberibacter asiaticus”
Authors: Wang Cheng , Fang Fang , Li Yun , Zhang Ling , Wu Jinghua , Li Tao , Zheng Yongqin , Xu Qian , Fan Shuting , Chen Jianchi , Deng Xiaoling , Zheng Zheng
Published: September 2022

“Candidatus Liberibacter asiaticus” (CLas) is the causal agent of citrus Huanglongbing (HLB, also called citrus greening disease), a highly destructive disease threatening citrus production worldwide. A novel Microviridae phage (named CLasMV1) has been found to infect CLas, providing a potential therapeutic strategy for CLas/HLB control. However, little is known about the CLasMV1 biology. In this study, we analyzed the population dynamics of CLasMV1 between the insect vector of CLas, the Asian citrus psyllid (ACP, Diaphorina citri Kuwayama) and the holoparasitic dodder plant (Cuscuta campestris Yunck.); both acquired CLasMV1-infected CLas from an HLB citrus. All CLas-positive dodder samples were CLasMV1-positive, whereas only 32% of CLas-positive ACP samples were identified as CLasMV1-positive. Quantitative analyses showed a similar distribution pattern of CLasMV1 phage and CLas among eight citrus cultivars by presenting at highest abundance in the fruit pith and/or the center axis of the fruit. Transcriptome analyses revealed the possible lytic activity of CLasMV1 on CLas in fruit pith as evidenced by high-level expressions of CLasMV1 genes, and CLas genes related to cell wall biogenesis and remodeling to maintain the CLas cell envelope integrity. The up-regulation of CLas genes were involved in restriction–modification system that could involve possible phage resistance for CLas during CLasMV1 infection. In addition, the regulation of CLas genes involved in cell surface components and Sec pathway by CLasMV1 phage could be beneficial for phage infection. This study expanded our knowledge of CLasMV1 phage that will benefit further CLas phage research and HLB control.
Novel Short PEG Chain-Substituted Porphyrins: Synthesis, Photochemistry, and In Vitro Photodynamic Activity against Cancer Cells
Authors: Lazewski Dawid , Kucinska Malgorzata , Potapskiy Edward , Kuzminska Joanna , Tezyk Artur , Popenda Lukasz , Jurga Stefan , Teubert Anna , Gdaniec Zofia , Kujawski Jacek , Grzyb Katarzyna , Pedzinski Tomasz , Murias Marek , Wierzchowski Marcin
Published: September 2022

This work presents the synthesis and characterization of metal-free, zinc (II), and cobalt (II) porphyrins substituted with short PEG chains. The synthesized compounds were characterized by UV-Vis, 1H and 13C NMR spectroscopy, and MALDI-TOF mass spectrometry. The origin of the absorption bands for tested compounds in the UV-Vis range was determined using a computational model based on the electron density functional theory (DFT) and its time-dependent variant (TD-DFT). The photosensitizing activity was evaluated by measuring the ability to generate singlet oxygen (ΦΔ), which reached values up to 0.54. The photodynamic activity was tested using bladder (5637), prostate (LNCaP), and melanoma (A375) cancer cell lines. In vitro experiments clearly showed the structure–activity relationship regarding types of substituents, their positions in the phenyl ring, and the variety of central metal ions on the porphyrin core. Notably, the metal-free derivative 3 and its zinc derivative 6 exerted strong cytotoxic activity toward 5637 cells, with IC50 values of 8 and 15 nM, respectively. None of the tested compounds induced a cytotoxic effect without irradiation. In conclusion, these results highlight the potential value of the tested compounds for PDT application.
Lumican, an Exerkine, Protects against Skeletal Muscle Loss
Authors: Cho Han Jin , Lee Young-Sun , Kim Da Ae , Moon Sung Ah , Lee Seung Eun , Lee Seung Hun , Koh Jung-Min
Published: September 2022

Exerkines are soluble factors secreted by exercised muscles, mimicking the effects of exercise in various organs, including the muscle itself. Lumican is reportedly secreted from muscles; however, its roles in skeletal muscle remain unknown. Herein, we found that lumican mRNA expression in the extensor digitorum longus was significantly higher in exercised mice than in unloading mice, and lumican stimulated myogenesis in vitro. Additionally, lumican knockdown significantly decreased muscle mass and cross-sectional area (CSA) of the muscle fiber in the gastrocnemius muscle of exercised mice. Lumican upregulated phosphorylation of p38 mitogen-activated protein kinase (MAPK) and a p38 inhibitor near completely blocked lumican-stimulated myogenesis. Inhibitors for integrin α2β1 and integrin ανβ3 also prevented lumican-stimulated myogenesis. Systemic lumican treatment, administered via the tail vein for 4 weeks, significantly increased relative muscle masses by 36.1% in ovariectomized mice. In addition, intramuscular lumican injection into unloaded muscles for 2 weeks significantly increased muscle mass by 8.5%. Both intravenous and intramuscular lumican treatment significantly increased muscle CSA. Our in vitro and in vivo experiments indicate that lumican is a muscle-secreted exerkine that affords protection against muscle loss by activating p38 MAPK via integrin receptors.
Regulation of Circadian Genes Nr1d1 and Nr1d2 in Sex-Different Manners during Liver Aging
Authors: Noh Sang Gyun , Jung Hee Jin , Kim Seungwoo , Arulkumar Radha , Kim Dae Hyun , Park Daeui , Chung Hae Young
Published: September 2022

Background: Circadian rhythm is associated with the aging process and sex differences; however, how age and sex can change circadian regulation systems remains unclear. Thus, we aimed to evaluate age- and sex-related changes in gene expression and identify sex-specific target molecules that can regulate aging. Methods: Rat livers were categorized into four groups, namely, young male, old male, young female, and old female, and the expression of several genes involved in the regulation of the circadian rhythm was confirmed by in silico and in vitro studies. Results: Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses showed that the expression of genes related to circadian rhythms changed more in males than in females during liver aging. In addition, differentially expressed gene analysis and quantitative real-time polymerase chain reaction/western blotting analysis revealed that Nr1d1 and Nr1d2 expression was upregulated in males during liver aging. Furthermore, the expression of other circadian genes, such as Arntl, Clock, Cry1/2, Per1/2, and Rora/c, decreased in males during liver aging; however, these genes showed various gene expression patterns in females during liver aging. Conclusions: Age-related elevation of Nr1d1/2 downregulates the expression of other circadian genes in males, but not females, during liver aging. Consequently, age-related upregulation of Nr1d1/2 may play a more crucial role in the change in circadian rhythms in males than in females during liver aging.
The Effect of Microcosm Biofilm Decontamination on Surface Topography, Chemistry, and Biocompatibility Dynamics of Implant Titanium Surfaces
Authors: Sousa Vanessa , Mardas Nikos , Spratt Dave , Hassan Iman A. , Walters Nick J. , Beltrán Víctor , Donos Nikolaos
Published: September 2022

Since the inception of dental implants, a steadily increasing prevalence of peri-implantitis has been documented. Irrespective of the treatment protocol applied for the management of peri-implantitis, this biofilm-associated pathology, continues to be a clinical challenge yielding unpredictable and variable levels of resolution, and in some cases resulting in implant loss. This paper investigated the effect of microcosm biofilm in vitro decontamination on surface topography, wettability, chemistry, and biocompatibility, following decontamination protocols applied to previously infected implant titanium (Ti) surfaces, both micro-rough -Sandblasted, Large-grit, Acid-etched (SLA)-and smooth surfaces -Machined (M). Microcosm biofilms were grown on SLA and M Ti discs. These were treated with TiBrushes (TiB), combination of TiB and photodynamic therapy (PDT), combination of TiB and 0.2%CHX/1%NaClO, plus or minus Ultraviolet-C (UV-C) radiation. Surface topography was evaluated by Scanning Electron Microscopy (SEM) and Laser Surface Profilometry. Surface function was analysed through wettability analysis. Surface chemistry evaluation of the discs was performed under SEM/Energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS). Biocompatibility was tested with the cytocompatibility assay using human osteoblast-like osteosarcoma cell line (MG-63) cells. Elemental analysis of the discs disclosed chemical surface alterations resulting from the different treatment modalities. Titanium, carbon, oxygen, sodium, aluminium, silver, were identified by EDX as the main components of all the discs. Based on the data drawn from this study, we have shown that following the decontamination of Ti surfaces the biomaterial surface chemistry and topography was altered. The type of treatment and Ti surface had a significant effect on cytocompatibility (p = 0.0001). Although, no treatment modality hindered the titanium surface biocompatibility, parameters such as the use of chemical agents and micro-rough surfaces had a higher cytotoxic effect in MG-63 cells. The use of smooth surfaces, and photofunctionalisation of the TiO2 layer had a beneficial effect on cytocompatibility following decontamination.
Ultrasound Neuromodulation Reduces Demyelination in a Rat Model of Multiple Sclerosis
Authors: Yang Feng-Yi , Huang Li-Hsin , Wu Meng-Ting , Pan Zih-Yun
Published: September 2022

Microglia, astrocytes, and oligodendrocyte progenitor cells (OPCs) may serve as targets for remyelination-enhancing therapy. Low-intensity pulsed ultrasound (LIPUS) has been demonstrated to ameliorate myelin loss and inhibit neuroinflammation in animal models of brain disorders; however, the underlying mechanisms through which LIPUS stimulates remyelination and glial activation are not well-understood. This study explored the impacts of LIPUS on remyelination and resident cells following lysolecithin (LPC)-induced local demyelination in the hippocampus. Demyelination was induced by the micro-injection of 1.5 μL of 1% LPC into the rat hippocampus, and the treatment groups received daily LIPUS stimulation for 5 days. The therapeutic effects of LIPUS on LPC-induced demyelination were assessed through immunohistochemistry staining. The staining was performed to evaluate remyelination and Iba-1 staining as a microglia marker. Our data revealed that LIPUS significantly increased myelin basic protein (MBP) expression. Moreover, the IHC results showed that LIPUS significantly inhibited glial cell activation, enhanced mature oligodendrocyte density, and promoted brain-derived neurotrophic factor (BDNF) expression at the lesion site. In addition, a heterologous population of microglia with various morphologies can be found in the demyelination lesion after LIPUS treatment. These data show that LIPUS stimulation may serve as a potential treatment for accelerating remyelination through the attenuation of glial activation and the enhancement of mature oligodendrocyte density and BDNF production.
Evaluation of a Serum-Free Medium for Human Epithelial and Stromal Cell Culture
Authors: Caneparo Christophe , Chabaud Stéphane , Fradette Julie , Bolduc Stéphane
Published: September 2022

Over the past decade, growing demand from many domains (research, cosmetics, pharmaceutical industries, etc.) has given rise to significant expansion of the number of in vitro cell cultures. Despite the widespread use of fetal bovine serum, many issues remain. Among them, the whole constitution of most serums remains unknown and is subject to significant variations. Furthermore, the presence of potential contamination and xenogeny elements is challenging for clinical applications, while limited production is an obstacle to the growing demand. To circumvent these issues, a Serum-Free Medium (SFM) has been developed to culture dermal and vesical fibroblasts and their corresponding epithelial cells, namely, keratinocytes and urothelial cells. To assess the impact of SFM on these cells, proliferation, clonogenic and metabolic assays have been compared over three passages to conditions associated with the use of a classic Fetal Bovine Serum-Containing Medium (FBSCM). The results showed that the SFM enabled fibroblast and epithelial cell proliferation while maintaining a morphology, cell size and metabolism similar to those of FBSCM. SFM has repeatedly been found to be better suited for epithelial cell proliferation and clonogenicity. Fibroblasts and epithelial cells also showed more significant mitochondrial metabolism in the SFM compared to the FBSCM condition. However, the SFM may need further optimization to improve fibroblast proliferation.
Novel Nanotechnology Approaches to Overcome Drug Resistance in the Treatment of Hepatocellular Carcinoma: Glypican 3 as a Useful Target for Innovative Therapies
Authors: Mossenta Monica , Busato Davide , Dal Bo Michele , Macor Paolo , Toffoli Giuseppe
Published: September 2022

Hepatocellular carcinoma (HCC) is the second most lethal tumor, with a 5-year survival rate of 18%. Early stage HCC is potentially treatable by therapies with curative intent, whereas chemoembolization/radioembolization and systemic therapies are the only therapeutic options for intermediate or advanced HCC. Drug resistance is a critical obstacle in the treatment of HCC that could be overcome by the use of targeted nanoparticle-based therapies directed towards specific tumor-associated antigens (TAAs) to improve drug delivery. Glypican 3 (GPC3) is a member of the glypican family, heparan sulfate proteoglycans bound to the cell surface via a glycosylphosphatidylinositol anchor. The high levels of GPC3 detected in HCC and the absence or very low levels in normal and non-malignant liver make GPC3 a promising TAA candidate for targeted nanoparticle-based therapies. The use of nanoparticles conjugated with anti-GPC3 agents may improve drug delivery, leading to a reduction in severe side effects caused by chemotherapy and increased drug release at the tumor site. In this review, we describe the main clinical features of HCC and the common treatment approaches. We propose the proteoglycan GPC3 as a useful TAA for targeted therapies. Finally, we describe nanotechnology approaches for anti-GPC3 drug delivery systems based on NPs for HCC treatment.
Transcription Factors Runx1 and Runx3 Suppress Keratin Expression in Undifferentiated Keratinocytes
Authors: Ogawa Eisaku , Edamitsu Tomohiro , Ohmori Hidetaka , Kohu Kazuyoshi , Kurokawa Mineo , Kiyonari Hiroshi , Satake Masanobu , Okuyama Ryuhei
Published: September 2022

The Runt-related transcription factor (Runx) family has been suggested to play roles in stem cell regulation, tissue development, and oncogenesis in various tissues/organs. In this study, we investigated the possible functions of Runx1 and Runx3 in keratinocyte differentiation. Both Runx1 and Runx3 proteins were detected in primary cultures of mouse keratinocytes. Proteins were localized in the nuclei of undifferentiated keratinocytes but translocated to the cytoplasm of differentiated cells. The siRNA-mediated inhibition of Runx1 and Runx3 expression increased expression of keratin 1 and keratin 10, which are early differentiation markers of keratinocytes. In contrast, overexpression of Runx1 and Runx3 suppressed keratin 1 and keratin 10 expression. Endogenous Runx1 and Runx3 proteins were associated with the promoter sequences of keratin 1 and keratin 10 genes in undifferentiated but not differentiated keratinocytes. In mouse skin, the inhibition of Runx1 and Runx3 expression by keratinocyte-specific gene targeting increased the ratios of keratin 1- and keratin 10-positive cells in the basal layer of the epidermis. On the other hand, inhibition of Runx1 and Runx3 expression did not alter the proliferation capacity of cultured or epidermal keratinocytes. These results suggest that Runx1 and Runx3 likely function to directly inhibit differentiation-induced expression of keratin 1 and keratin 10 genes but are not involved in the regulation of keratinocyte proliferation.
Silica Shell Thickness-Dependent Fluorescence Properties of SiO2@Ag@SiO2@QDs Nanocomposites
Authors: Hahm Eunil , Jo Ahla , Lee Sang Hun , Kang Homan , Pham Xuan-Hung , Jun Bong-Hyun
Published: September 2022

Silica shell coatings, which constitute important technology for nanoparticle (NP) developments, are utilized in many applications. The silica shell’s thickness greatly affects distance-dependent optical properties, such as metal-enhanced fluorescence (MEF) and fluorescence quenching in plasmonic nanocomposites. However, the precise control of silica-shell thicknesses has been mainly conducted on single metal NPs, and rarely on complex nanocomposites. In this study, silica shell-coated Ag nanoparticle-assembled silica nanoparticles (SiO2@Ag@SiO2), with finely controlled silica shell thicknesses (4 nm to 38 nm), were prepared, and quantum dots (QDs) were introduced onto SiO2@Ag@SiO2. The dominant effect between plasmonic quenching and MEF was defined depending on the thickness of the silica shell between Ag and QDs. When the distance between Ag NPs to QDs was less than ~10 nm, SiO2@Ag@SiO2@QDs showed weaker fluorescence intensities than SiO2@QD (without metal) due to the quenching effect. On the other hand, when the distance between Ag NPs to QDs was from 10 nm to 14 nm, the fluorescence intensity of SiO2@Ag@SiO2@QD was stronger than SiO2@QDs due to MEF. The results provide background knowledge for controlling the thickness of silica shells in metal-containing nanocomposites and facilitate the development of potential applications utilizing the optimal plasmonic phenomenon.
Optimization of Signal Peptide via Site-Directed Mutagenesis for Enhanced Secretion of Heterologous Proteins in Lactococcus lactis
Authors: Alias Nur Aqlili Riana , Song Adelene Ai-Lian , Alitheen Noorjahan Banu , Rahim Raha Abdul , Othman Siti Sarah , In Lionel Lian Aun
Published: September 2022

Secretion efficiency of heterologous proteins in the Generally Regarded As Safe (GRAS) Lactococcus lactis is often reported to be insufficiently low due to limitations such as poor targeting and translocation by the signal peptide or degradation by the host proteases. In this study, the secretion efficiency in the host was enhanced through the utilization of a heterologous signal peptide (SP) SPK1 of Pediococcus pentosaceus. The SPK1 was subjected to site-directed mutations targeting its tripartite N-, H-, and C-domains, and the effect on secretion efficiency as compared to the wild-type SPK1 and native lactococcal USP45 was determined on a reporter nuclease (NUC) of Staphylococcus aureus. A Fluorescence Resonance Energy Transfer (FRET) analysis indicated that four out of eight SPK1 variants successfully enhanced the secretion of NUC, with the best mutant, SPKM19, showing elevated secretion efficiency up to 88% (or by 1.4-fold) and an improved secretion activity yield of 0.292 ± 0.122 U/mL (or by 1.7-fold) compared to the wild-type SPK1. Modifications of the SPK1 at the cleavage site C-domain region had successfully augmented the secretion efficiency. Meanwhile, mutations in the H-domain region had resulted in a detrimental effect on the NUC secretion. The development of heterologous SPs with better efficacy than the USP45 has been demonstrated in this study for enhanced secretion of heterologous production and mucosal delivery applications in the lactococcal host.
Role of Nitric Oxide in Postharvest Senescence of Fruits
Authors: Li Changxia , Yu Wenjin , Liao Weibiao
Published: September 2022

Nitric oxide (NO) acts as a gaseous signalling molecule and is considered to be a key regulator in the postharvest storage of fruits. Postharvest senescence is one of the most serious threats affecting the usage and economic value of fruits. Most recent studies have found that exogenous NO application can effectively improve the quality and prolong the shelf life of fruit postharvest by inhibiting postharvest diseases and alleviating chilling injury. Understanding the roles of NO is essential to elucidating how NO activates the appropriate set of responses to postharvest senescence. Here, we concluded that exogenous NO treatment alleviated senescence in postharvest fruit and attributed this to the following factors: (1) ethylene biosynthesis, (2) the antioxidant system, (3) polyamine metabolism and γ-aminobutyric acid (GABA) shunting, (4) cell wall metabolism, (5) sugar metabolism, (6) energy metabolism, (7) the CRT/DRE-binding factor (CBF) pathway and (8) S-nitrosylation. Moreover, crosstalk between NO and hydrogen sulfide (H2S), hydrogen peroxide (H2O2), oxalic acid (OA), arginine (Arg), GATA or plant hormone abscisic acid (ABA), melatonin (MT), and methyl jasmonate (MeJA), along with the regulation of key genes, were found to be very important in responses to postharvest senescence. In this study, we focus on the recent knowledge concerning the alleviative effect of NO on postharvest senescence, covering ethylene biosynthesis, the antioxidant system and related gene and protein expression.
Ligand-Based Virtual Screening and Molecular Docking of Benzimidazoles as Potential Inhibitors of Triosephosphate Isomerase Identified New Trypanocidal Agents
Authors: Vázquez-Jiménez Lenci K. , Juárez-Saldivar Alfredo , Gómez-Escobedo Rogelio , Delgado-Maldonado Timoteo , Méndez-Álvarez Domingo , Palos Isidro , Bandyopadhyay Debasish , Gaona-Lopez Carlos , Ortiz-Pérez Eyra , Nogueda-Torres Benjamín , Ramírez-Moreno Esther , Rivera Gildardo
Published: September 2022

Trypanosoma cruzi (T. cruzi) is a parasite that affects humans and other mammals. T. cruzi depends on glycolysis as a source of adenosine triphosphate (ATP) supply, and triosephosphate isomerase (TIM) plays a key role in this metabolic pathway. This enzyme is an attractive target for the design of new trypanocidal drugs. In this study, a ligand-based virtual screening (LBVS) from the ZINC15 database using benzimidazole as a scaffold was accomplished. Later, a molecular docking on the interface of T. cruzi TIM (TcTIM) was performed and the compounds were grouped by interaction profiles. Subsequently, a selection of compounds was made based on cost and availability for in vitro evaluation against blood trypomastigotes. Finally, the compounds were analyzed by molecular dynamics simulation, and physicochemical and pharmacokinetic properties were determined using SwissADME software. A total of 1604 molecules were obtained as potential TcTIM inhibitors. BP2 and BP5 showed trypanocidal activity with half-maximal lytic concentration (LC50) values of 155.86 and 226.30 µM, respectively. Molecular docking and molecular dynamics simulation analyzes showed a favorable docking score of BP5 compound on TcTIM. Additionally, BP5 showed a low docking score (−5.9 Kcal/mol) on human TIM compared to the control ligand (−7.2 Kcal/mol). Both compounds BP2 and BP5 showed good physicochemical and pharmacokinetic properties as new anti-T. cruzi agents.
Development of Methotrexate Complexes Endowed with New Biological Properties Envisioned for Musculoskeletal Regeneration in Rheumatoid Arthritis Environments
Authors: Fernández-Villa Daniel , Ramírez-Jiménez Rosa Ana , Aranaz Inmaculada , Acosta Niuris , Vázquez-Lasa Blanca , Rojo Luis
Published: September 2022

Methotrexate (MTX) administration is the gold standard treatment for rheumatoid arthritis (RA), but its effects are limited to preventing the progression of the disease. Therefore, effective regenerative therapies for damaged tissues are still to be developed. In this regard, MTX complexes of general molecular formula M(MTX)·xH2O, where M = Sr, Zn, or Mg, were synthesized and physicochemically characterized by TGA, XRD, NMR, ATR–FTIR, and EDAX spectroscopies. Characterization results demonstrated the coordination between the different cations and MTX via two monodentate bonds with the carboxylate groups of MTX. Cation complexation provided MTX with new bioactive properties such as increasing the deposition of glycosaminoglycans (GAGs) and alternative anti-inflammatory capacities, without compromising the immunosuppressant properties of MTX on macrophages. Lastly, these new complexes were loaded into spray-dried chitosan microparticles as a proof of concept that they can be encapsulated and further delivered in situ in RA-affected joints, envisioning them as a suitable alternative to oral MTX therapy.
Genome-Wide Identification and Expression Patterns of the SWEET Gene Family in Bletilla striata and its Responses to Low Temperature and Oxidative Stress
Authors: Lu Chan , Ye Jun , Chang Yuanqing , Mi Zeyuan , Liu Shuai , Wang Donghao , Wang Zhezhi , Niu Junfeng
Published: September 2022

SWEETs (sugars will eventually be exported transporters), a well-known class of sugar transporters, are involved in plant growth and development, sugar transport, biotic and abiotic stresses, etc. However, to date, there have been few investigations of SWEETs in Orchidaceae. In this study, 23 SWEET genes were identified in Bletilla striata for the first time, with an MtN3/saliva conserved domain, and were divided into four subgroups by phylogenetic tree. The same subfamily members had similar gene structures and motifs. Multiple cis-elements related to sugar and environmental stresses were found in the promoter region. Further, 21 genes were localized on 11 chromosomes and 2 paralogous pairs were found via intraspecific collinearity analysis. Expression profiling results showed that BsSWEETs were tissue-specific. It also revealed that BsSWEET10 and BsSWEET18 were responsive to low temperature and oxidative stresses. In addition, subcellular localization study indicated that BsSWEET15 and BsSWEET16 were localized in the cell membrane. This study provided important clues for the in-depth elucidation of the sugar transport mechanism of BsSWEET genes and their functional roles in response to abiotic stresses.
Effect of Calcium-Sulphate-Bicarbonate Water in a Murine Model of Non-Alcoholic Fatty Liver Disease: A Histopathology Study
Authors: Carpino Guido , Overi Diletta , Onori Paolo , Franchitto Antonio , Cardinale Vincenzo , Alvaro Domenico , Gaudio Eugenio
Published: September 2022

The progression of nonalcoholic fatty liver disease (NAFLD) is associated with alterations of the gut–liver axis. The activation of toll-like receptor 4 (TLR4) pathways by endotoxins, such as lipopolysaccharide (LPS), contributes to liver injury. The aim of the present study was to evaluate the possible beneficial effects of a calcium-sulphate-bicarbonate natural mineral water on the gut–liver axis by evaluating liver and terminal ileum histopathology in a murine model of NAFLD. NAFLD was induced in mice by administrating a methionine-choline-deficient (MCD) diet. The following experimental groups were evaluated: controls (N = 10); MCD+Tap water (MCD; N = 10); MCD+Calcium-sulphate-bicarbonate water (MCD/Wcsb; N = 10). Mice were euthanised after 4 and 8 weeks. Liver and terminal ileum samples were collected. Samples were studied by histomorphology, immunohistochemistry, and immunofluorescence. In mice subjected to the MCD diet, treatment with mineral water improved inflammation and fibrosis, and was associated with a reduced number of activated hepatic stellate cells when compared to MCD mice not treated with mineral water. Moreover, MCD/Wcsb mice showed lower liver LPS localization and less activation of TLR4 pathways compared to the MCD. Finally, Wcsb treatment was associated with improved histopathology and higher occludin positivity in intestinal mucosa. In conclusion, calcium-sulphate-bicarbonate water may exert modulatory activity on the gut–liver axis in MCD mice, suggesting potential beneficial effects on NAFLD.