bluuebunny/crossref_metadata_2025 huggingface dataset:
A Strategy for Rapid Acquisition of the β-D-Fructofuranosidase Gene through Chemical Synthesis and New Function of Its Encoded Enzyme to Improve Gel Properties during Yogurt Processing
Authors: Chen Zhou , Shen Yimei , Xu Jiangqi
Published: April 2023

A chemical gene synthesis strategy was developed in order to obtain β-D-fructofuranosidase, and a novel gene, AlFFase3, was characterized from Aspergillus luchuensis and expressed in Escherichia coli. The recombinant protein was purified, showing a molecular mass of 68.0 kDa on SDS-PAGE, and displaying a specific activity towards sucrose of up to 771.2 U mg−1, indicating its exceptional enzymatic capacity. AlFFase3 exhibited stability between pH 5.5 and 7.5, with maximal activity at pH 6.5 and 40 °C. Impressively, AlFFase3, as a soluble protein, was resistant to digestion by various common proteases, including Flavourzyme, acidic protease, pepsin, neutral protease, Proteinase K, alkaline proteinase, and trypsin. AlFFase3 also demonstrated significant transfructosylation activity, with a yield of various fructooligosaccharides up to 67%, higher than almost all other reports. Furthermore, we demonstrated that the addition of AlFFase3 enhanced the growth of probiotics in yogurt, thereby increasing its nutritional value. AlFFase3 also improved the formation of yogurt gel, reducing the gel formation time and lowering the elasticity while increasing its viscosity, thereby improving the palatability of yogurt and reducing production costs.
Evaluation of Sufu Fermented Using Mucor racemosus M2: Biochemical, Textural, Structural and Microbiological Properties
Authors: Xie Yuan , Guan Ziyu , Zhang Shitong , Zhang Jie , Yang Zhihui , Regenstein Joe M. , Zhou Peng
Published: April 2023

The quality and safety of sufu fermented using Mucor racemosa M2 was studied and compared with naturally fermented sufu. After 90 days post-fermentation, both naturally fermented and inoculated fermented sufu reached the maturity standard of sufu, and the degree of protein hydrolysis of natural sufu (WP/TP: 34% ± 1%; AAN/TN: 33% ± 1%) was slightly higher than that of the inoculated sufu (WP/TP: 28.2% ± 0.4%; AAN/TN: 27% ± 1%). The hardness and adhesiveness of inoculated sufu (Hadness: 1063 g ± 211 g; Adhesiveness: −80 g ± 47 g) were significantly greater than those of natural sufu (Hadness: 790 g ± 57 g; Adhesiveness: −23 g ± 28 g), and the internal structure of natural sufu was denser and more uniform than that of inoculated sufu. A total of 50 aroma compounds were detected in natural and inoculated sufu. The total number of bacterial colonies in naturally fermented sufu was significantly higher than that in inoculated sufu, and the pathogenic bacteria in both types of fermented sufu were lower than the limit of pathogenic bacteria required in fermented soybean products. The content of biogenic amines in sufu was determined by high performance liquid chromatography (HPLC), and the results showed that the content of biogenic amines (Putrescine, Cadaverine, Histamine, Tyramine, etc.) in naturally fermented sufu was significantly higher than that in inoculated fermented sufu. Especially the histamine content, after 90 days of fermentation, was found to be 64.95 ± 4.55 for inoculated fertilization and 44.24 ± 0.71 for natural fertilization. Overall, the quality of inoculated sufu was somewhat better than that of natural sufu, and the M2 strain can be used to ferment sufu.
Applicable Strains, Processing Techniques and Health Benefits of Fermented Oat Beverages: A Review
Authors: Yu Qian , Qian Jiaqin , Guo Yahui , Qian He , Yao Weirong , Cheng Yuliang
Published: April 2023

Based on the high nutrients of oat and the demand of health-conscious consumers for value-added and functional foods, fermented oat beverages have great market prospects. This review summarizes the applicable strains, processing techniques and health benefits of fermented oat beverages. Firstly, the fermentation characteristics and conditions of the applicable strains are systematically described. Secondly, the advantages of pre-treatment processes such as enzymatic hydrolysis, germination, milling and drying are summarized. Furthermore, fermented oat beverages can increase the nutrient content and reduce the content of anti-nutritional factors, thereby reducing some risk factors related to many diseases such as diabetes, high cholesterol and high blood pressure. This paper discusses the current research status of fermented oat beverages, which has academic significance for researchers interested in the application potential of oat. Future studies on fermenting oat beverages can focus on the development of special compound fermentation agents and the richness of their taste.
A Light Scattering Investigation of Enzymatic Gelation in Self-Assembling Peptides
Authors: Buzzaccaro Stefano , Ruzzi Vincenzo , Gelain Fabrizio , Piazza Roberto
Published: April 2023

Self-assembling peptides (SAPs) have been increasingly studied as hydrogel–former gelators because they can create biocompatible environments. A common strategy to trigger gelation, is to use a pH variation, but most methods result in a change in pH that is too rapid, leading to gels with hardly reproducible properties. Here, we use the urea–urease reaction to tune gel properties, by a slow and uniform pH increase. We were able to produce very homogeneous and transparent gels at several SAP concentrations, ranging from c=1g/L to c=10g/L. In addition, by exploiting such a pH control strategy, and combining photon correlation imaging with dynamic light scattering measurements, we managed to unravel the mechanism by which gelation occurs in solutions of (LDLK)3-based SAPs. We found that, in diluted and concentrated solutions, gelation follows different pathways. This leads to gels with different microscopic dynamics and capability of trapping nanoparticles. At high concentrations, a strong gel is formed, made of relatively thick and rigid branches that firmly entrap nanoparticles. By contrast, the gel formed in dilute conditions is weaker, characterized by entanglements and crosslinks of very thin and flexible filaments. The gel is still able to entrap nanoparticles, but their motion is not completely arrested. These different gel morphologies can potentially be exploited for controlled multiple drug release.
Early Feasibility of an Activity-Based Intervention for Improving Ingestive Functions in Older Adults with Oropharyngeal Dysphagia
Authors: Hansen Tina , Laursen Louise Bolvig , Hansen Maria Swennergren
Published: April 2023

There is growing awareness about the use of combined strength- and skill-based swallowing training for improving swallowing physiology in the event of dysphagia. Such an approach involves focusing on coordination and timing as well as swallowing strengthening in the context of increased exercise complexity in eating and drinking activities. This study aimed to determine the early feasibility of a newly developed 12-week intervention, named the ACT-ING program (ACTivity-based strength and skill training of swallowing to improve INGestion), in older adults with dysphagia and generalized sarcopenia. In a multiple-case-study design, seven participants above 65 years of age (five women and two men) with slight to severe dysphagia and indications of sarcopenia underwent the intervention during hospitalization and in the community after discharge. The ACT-ING program met most of the feasibility marks in terms of demand (73.3% of those invited accepted participation), safety (100%), no reports of adverse events, tolerance (85.7%), usability (100%), and acceptability (100%). Three putative mediators of change (experienced autonomy support, in-therapy engagement, and perceived improvement in swallowing capacity) appeared to have been best accomplished in participants with slight to moderate dysphagia. The ACT-ING program showed preliminary evidence of early feasibility, warranting further early-phase dose articulation and proof-of-concept trials.
Positive Impact of Home ERT for Mucopolysaccharidoses and Pompe Disease: The Lesson Learnt from the COVID-19 Pandemic
Authors: Fiumara Agata , Lanzafame Giuseppina , Sapuppo Annamaria , Arena Alessia , Cirnigliaro Lara , Barone Rita
Published: April 2023

Objective: Patients with Lysosomal disorders (LSDs) are treated with regular infusions of enzyme replacement therapy (ERT). During the COVID-19 pandemic, home treatment was permitted. This study aimed at monitoring the patients’ compliance with home therapy and its effects on physical, psychological, and relational issues. Moreover, we also tested the possible impact of home therapy on familial relationships and contacts with the referral hospital. Materials and Methods: Thirteen patients with Pompe disease (N = 8) and MPS (N = 5) were tested through an online questionnaire designed to assess their level of appreciation and satisfaction with home therapy and their feelings about the referral centre and psychological support provided. Results: Most of the patients (84%) stressed the positive impact of home therapy. All patients described a significant reduction in stressful conditions associated with the need to attend the hospital every week or two. Conclusions: Home ERT leads to a clear improvement in “daily life skills”, as represented in our by sample by positive feelings, better emotional self-control, and an increased ability to understand the feelings of relatives. Our data underline the paramount positive effect home ERT has on both patients and their families.
Opportunities and Threats of the Legally Facilitated Performance-Based Managed Entry Agreements in Slovakia: The Early-Adoption Perspective
Authors: Hospodková Petra , Gilíková Klára , Barták Miroslav , Marušáková Elena , Tichopád Aleš
Published: April 2023

Slovakia has adopted an amendment to Act No. 363/2011, regulating, among other things, drug reimbursement and is undergoing a significant change in the availability of innovative treatments for patients. High expectations are associated with arrangements related to performance-based managed entry agreements. Opinions and positions towards this change appear to be inconsistent, and for the further application of the law in practice and when setting up the main implementation processes, it is necessary to understand the positions and opinions of the individual actors who are involved in the PB-MEA process. The interviews were conducted in the period from 20 May to 15 August 2022 around the same time as the finalisation of the amendment to Act No. 363/2011 and its adoption. A roughly one-hour open interview was conducted on a sample of 12 stakeholders in the following groups: representatives of the Ministry of Health, health-care providers, pharmaceutical companies and others, including a health insurance company. The main objective was to qualitatively describe the perception of this topic by key stakeholders in Slovakia. The responses were analysed using MAXQDATA 2022 software to obtain codes associated with key expressions. We identified three main strong top categories of expressions that strongly dominated the pro-management interviews with stakeholders: legislation, opportunities and threats. Ambiguity and insufficient coverage of the new law, improved availability of medicinal products and threats associated with data, IT systems and potentially unfavourable new reimbursement schemes were identified as key topics of each of the said top categories, respectively. Among individual sets of respondents, there is frequent consensus on both opportunities and threats in the area of implementing process changes in PB-MEA. For the successful implementation of the law in practice, some basic threats need to be removed, among which in particular is insufficient data infrastructure.
Effect of Magnetic Water Treatment on the Growth, Nutritional Status, and Yield of Lettuce Plants with Irrigation Rate
Authors: Putti Fernando Ferrari , Vicente Eduardo Festozo , Chaves Prínscilla Pâmela Nunes , Mantoan Luís Paulo Benetti , Cremasco Camila Pires , Arruda Bruna , Forti Juliane Cristina , Junior Josué Ferreira Silva , Campos Marcelo , Reis André Rodrigues dos , Filho Luís Roberto Almeida Gabriel
Published: April 2023

Climate change is causing an increase in dry spells, altering rainfall patterns and soil moisture, and affecting water and nutrient plant uptake, which inevitably affects vegetable production. To mitigate this issue, some technologies that allow the maintenance of the ideal soil moisture for the uptake process are being investigated. Considering this, we hypothesize that the use of water treated with a magnetic field can increase water use efficiency in lettuce crop production. Thus, the present study aimed to evaluate the effect of the irrigation rate of magnetically treated water on biomass accumulation and nutrient uptake by lettuce plants. An experiment was conducted in a randomized block design with a 2 × 5 factorial arrangement of two water sources (conventional water and magnetically treated water) and five irrigation application rates to replace crop evaporation: 25, 50, 75, 100, and 125%, with five replicates. The use of magnetically treated water increased the concentrations of nitrogen and phosphorus in leaves, meaning that it induced higher nitrogen assimilation, leading to increases in agronomical characteristics (leaf number, fresh and dry shoot weight, fresh and dry root weight). The conclusions of this study showcase that magnetically treated water has beneficial effects on lettuce plants, improving their nutritional status and yield. Moreover, the results presented can lead to an increase in water use efficiency, thus optimizing irrigation management.
Applicability of Variable-Rate Nitrogen Top Dressing Based on Measurement of the Within-Field Variability of Soil Nutrients for Cabbage Production
Authors: Nakano Yuka , Noda Shintaro , Miyake Yasunari , Kogoshi Masayuki , Sato Fumio , Iijima Wataru
Published: April 2023

To improve the efficiency of nitrogen (N) fertilization, it is necessary to perform rapid direct measurements in the field rather than time-consuming laboratory-based chemical analysis. Herein, crop and soil data from the early stages of cabbage growth were acquired through two fall cultivations. Chlorophyll meter value, height, and projected leaf area were evaluated as crop indicators. A positive correlation was observed between the projected leaf area or its rate of increase 2–3 weeks after transplantation and head fresh weight (FW). After comparing two water-content reflectometers (WCR) and a nitrate sensor, we selected a WCR with a 12 cm-long rod as the soil indicator. The diagnostic method was verified using varying amounts of N basal fertilizer during spring cultivation. The variable rate of N top dressing (25, 50, and 75% total N) based on the electrical conductivity (EC) 14 days after transplantation reduced the subsequent EC variability. No differences in head FW were observed between the treatments. A 25% reduction in N fertilizer was considered possible for half of the plots. The quantity of inorganic N extracted by potassium chloride from the crop soil after cultivation was unaffected by the amount of N fertilizer. Therefore, the diagnostic method proposed herein is suitable for soil N management.
Physiological and Nutritional Responses of Ungrafted Merlot and Cabernet Sauvignon Vines or Grafted to 101-14 Mgt and 1103P Rootstocks Exposed to an Excess of Boron
Authors: Nikolaou Kleopatra-Eleni , Chatzistathis Theocharis , Theocharis Serafeim , Argiriou Anagnostis , Koundouras Stefanos
Published: April 2023

The aim of this study is to analyze the effect of excess boron (B) on the nutrient uptake, growth, and physiological performance of grapevines. Merlot and Cabernet Franc grapevines, either own-rooted or grafted onto 1103P and 101-14 Mgt rootstocks, were exposed in a hydroponic pot experiment to 0.5 mM boron for sixty days. Twenty-five days following the beginning of B treatment, the first symptoms of boron toxicity appeared, including leaf edge and margin yellowing, subsequent necrosis, and cupping of leaf blades. At harvest, sixty days after the start of the experiment, B concentration of the treated vines increased in all parts of the vine in the following order: Leaves > Roots > Trunks > Shoots. Leaf Boron concentration in treated vines ranged from 980.67 to 1064.37 mg kg−1 d.w. Boron excess significantly reduced the concentrations of all macro and micronutrients studied in this experiment. The total leaf chlorophyll (Chl) concentration decreased from 35.46 to 44.45%, thirty and sixty days, respectively, from the beginning of the boron treatments. In addition, an excess amount of boron resulted in a dramatic decrease in net CO2 assimilation rate, stem water potential, and PSII maximum quantum yield, irrespective of vine type (own-rooted or grafted). At the end of the experimental period, the total leaf phenolic content increased by 71.73% in Merlot and by 71.16% in Cabernet Franc due to Boron stress. The tressed vines grafted onto 1103P showed increased shoot and root dry weights, leaf chlorophyll content, CO2 assimilation rates, and Fv/Fm ratio compared to vines grafted onto 101-14 Mgt. It was found that rootstocks play an important role in B toxicity. The results showed that the leaf accumulation of boron was delayed to a greater extent by 1103P rootstock compared to 101-14 Mgt, resulting in the earlier appearance of leaf toxicity symptoms in vines grafted onto 101-14 Mgt rootstock compared to 1103P.
Housing Supply and How It Is Related to Social Inequalities—Air Pollution, Green Spaces, Crime Levels, and Poor Areas—In Catalonia
Authors: Perafita Xavier , Saez Marc
Published: April 2023

We carried out a search of over 12,000 houses offered on the rental market in Catalonia and assessed the possibility of families below the poverty threshold being able to rent these homes. In this regard, we wanted to evaluate whether the economic situation of families is able to influence their social environment, surroundings, and safety. We observed how their economic situation can allow families the possibility of developing a life without exposure to health risks, and how economic constraints result in disadvantages in several areas of life. The results show how families at risk of poverty live in less favourable conditions and experience a widening of different gaps, with current prices leading to a possible poverty trap for the most disadvantaged groups. The higher the percentage of the population below the threshold, the lower the possibility of not being able to rent a house compared to areas with a lower prevalence of population below the threshold. This association was observed both when considering the risk linearly and non-linearly. Linearly, the probability of not renting a house was reduced by 8.36% for each 1% increase in the prevalence of population at risk of extreme poverty. In the second, third and fourth percentage quartiles, the probability of not being able to rent a house decreased by 21.13%, 48.61%, and 57.79%, respectively. In addition, the effect was different inside and outside of metropolitan areas, with the former showing a decrease of 19.05% in the probability of renting a house, whereas outside metropolitan areas the probability increased by 5.70%.
International Music Therapists’ Perceptions and Experiences in Telehealth Music Therapy Provision
Authors: Clements-Cortés Amy , Pranjić Marija , Knott David , Mercadal-Brotons Melissa , Fuller Allison , Kelly Lisa , Selvarajah Indra , Vaudreuil Rebecca
Published: April 2023

The use of telehealth within music therapy practice has increased through necessity in recent years. To contribute to the evolving evidence base, this current study on Telehealth Music Therapy (TMT) was undertaken to investigate the telehealth provision experiences of music therapists internationally. Participants completed an anonymous online cross-sectional survey covering demographics, clinical practice, telehealth provision, and telehealth perceptions. Descriptive and inferential statistics, in combination with thematic analysis, were used to analyze the data. A total of 572 music therapists from 29 countries experienced in providing TMT took part in this study. The results showed that the overall number of clinical hours (TMT and in-person hours combined) declined due to the pandemic. Participants also reported reduced perceived success rates in utilizing both live and pre-recorded music in TMT sessions when compared to in-person sessions. Although many music therapists rose to the challenges posed by the pandemic by incorporating TMT delivery modes, there was no clear agreement on whether TMT has more benefits than drawbacks; however, reported benefits included increased client access and caregiver involvement. Furthermore, a correlation analysis revealed moderate-to-strong positive associations between respondents who perceived TMT to have more benefits than drawbacks, proficiency at administering assessments over telehealth, and perceived likelihood of using telehealth in the future. Regarding the influence of primary theoretical orientation and work setting, respondents who selected music psychotherapy as a primary theoretical orientation had more experience providing TMT prior to the pandemic while those primarily working in private practice were most inclined to continue TMT services post-pandemic. Benefits and drawbacks are discussed and future recommendations for TMT are provided.
Infantile Hemangioma: A Common Lesion in a Vulnerable Population
Authors: Nazemian Samon , Sharif Shohreh , Childers Esther L. B.
Published: April 2023

Infantile hemangioma (IH) is important to all dentists, particularly dentists providing services to pediatric populations, because significant morbidity and mortality can occur from vascular lesions in children. Specialists of the oral cavity have the responsibility to identify patients with IH, a lesion that could be potentially life threatening. We present a case of IH and will provide a narrative review of the most recent literature. We discuss the diagnosis, risk stratification, treatment, complications, and impact on routine dental procedures. A proper diagnosis is crucial as oral and perioral IH are considered high-risk due to the increased risk of ulceration and feeding impairment. Referral to a hemangioma specialist for comprehensive team treatment is optimal. The natural history of IH consists of a long proliferative phase, which manifests as clinically visible growth. As a result of early encounters with patients, the pediatric dentist may often be considered the primary care provider.
Experiences of People with Cardiovascular Disease during COVID-19 in Sweden: A Qualitative Study
Authors: Sjödahl Hammarlund Catharina , Norlander Anna , Brogårdh Christina
Published: April 2023

Although people with cardiovascular conditions were subjected to more rigorous restrictions during the COVID-19 pandemic, there is limited knowledge of how the restrictions affected their lives and well-being. Thus, the aim of this study was to describe how people with cardiovascular conditions experienced their life situation and physical and mental health during the second wave of the pandemic in Sweden. Fifteen participants (median age 69 years; nine women) were individually interviewed, and data were analyzed with systematic text condensation. The findings revealed that some of the participants were fearful of contracting COVID-19 as their medical condition made them vulnerable. Additionally, the restrictions changed their daily routines and their ability to take part in social activities, as well as their access to specialized outpatient care (medical check-ups and physiotherapy). Although emotional and psychological distress were present, several participants found strategies that reduced their worries, such as exercising and meeting friends outdoors. However, some had adopted a more sedentary lifestyle and unhealthy diets. These findings indicate that healthcare professionals should provide individualized support to persons with cardiovascular diseases in order to find well-functioning emotion- and problem-focused strategies aimed at improving physical and mental health during crises such as pandemics.
Efficient Algorithm for Constructing Order K Voronoi Diagrams in Road Networks
Authors: Chen Bi Yu , Huang Huihuang , Chen Hui-Ping , Liu Wenxuan , Chen Xuan-Yan , Jia Tao
Published: April 2023

The order k Voronoi diagram (OkVD) is an effective geometric construction to partition the geographical space into a set of Voronoi regions such that all locations within a Voronoi region share the same k nearest points of interest (POIs). Despite the broad applications of OkVD in various geographical analysis, few efficient algorithms have been proposed to construct OkVD in real road networks. This study proposes a novel algorithm consisting of two stages. In the first stage, a new one-to-all k shortest path finding procedure is proposed to efficiently determine the shortest paths to k nearest POIs for each node. In the second stage, a new recursive procedure is introduced to effectively divide boundary links within different Voronoi regions using the hierarchical tessellation property of the OkVD. To demonstrate the applicability of the proposed OkVD construction algorithm, a case study of place-based accessibility evaluation is carried out. Computational experiments are also conducted on five real road networks with different sizes, and results show that the proposed OkVD algorithm performed significantly better than state-of-the-art algorithms.
MKK4 Inhibitors—Recent Development Status and Therapeutic Potential
Authors: Katzengruber Leon , Sander Pascal , Laufer Stefan
Published: April 2023

MKK4 (mitogen-activated protein kinase kinase 4; also referred to as MEK4) is a dual-specificity protein kinase that phosphorylates and regulates both JNK (c-Jun N-terminal kinase) and p38 MAPK (p38 mitogen-activated protein kinase) signaling pathways and therefore has a great impact on cell proliferation, differentiation and apoptosis. Overexpression of MKK4 has been associated with aggressive cancer types, including metastatic prostate and ovarian cancer and triple-negative breast cancer. In addition, MKK4 has been identified as a key regulator in liver regeneration. Therefore, MKK4 is a promising target both for cancer therapeutics and for the treatment of liver-associated diseases, offering an alternative to liver transplantation. The recent reports on new inhibitors, as well as the formation of a startup company investigating an inhibitor in clinical trials, show the importance and interest of MKK4 in drug discovery. In this review, we highlight the significance of MKK4 in cancer development and other diseases, as well as its unique role in liver regeneration. Furthermore, we present the most recent progress in MKK4 drug discovery and future challenges in the development of MKK4-targeting drugs.
Targeting the Sequences of Circulating Tumor DNA of Cholangiocarcinomas and Its Applications and Limitations in Clinical Practice
Authors: Kim Kyung-Hee , Yi Hyon-Seung , Lee Hyunjung , Bae Go-Eun , Yeo Min-Kyung
Published: April 2023

Cholangiocarcinoma is a malignant epithelial tumor arising from bile ducts that is frequently fatal. Diagnosis is difficult due to tumor location in the biliary tract. Earlier diagnosis requires less invasive methods of identifying effective biomarkers for cholangiocarcinoma. The present study investigated the genomic profiles of cell-free DNA (cfDNA) and DNA from corresponding primary cholangiocarcinomas using a targeted sequencing panel. Somatic mutations in primary tumor DNA and circulating tumor DNA (ctDNA) were compared and clinical applications of ctDNA validated in patients with cholangiocarcinoma. A comparison of primary tumor DNA and ctDNA identified somatic mutations in patients with early cholangiocarcinomas that showed clinical feasibility for early screening. The predictive value of single-nucleotide variants (SNVs) of preoperative plasma cfDNA positive for somatic mutations of the primary tumor was 42%. The sensitivity and specificity of postoperative plasma SNVs in detecting clinical recurrence were 44% and 45%, respectively. Targetable fibroblast growth factor receptor 2 (FGFR2) and Kirsten rat sarcoma virus (KRAS) mutations were detected in 5% of ctDNA samples from patients with cholangiocarcinoma. These findings showed that genomic profiling of cfDNA was useful in clinical evaluation, although ctDNA had limited ability to detect mutations in cholangiocarcinoma patients. Serial monitoring of ctDNA is important clinically and in assessing real-time molecular aberrations in cholangiocarcinoma patients.
Evidence of Disruption in Neural Regeneration in Dry Eye Secondary to Rheumatoid Arthritis
Authors: Sonkodi Balázs , Csorba Anita , Marsovszky László , Balog Attila , Kopper Bence , Nagy Zoltán Zsolt , Resch Miklós D.
Published: April 2023

The purpose of our study was to analyze abnormal neural regeneration activity in the cornea through means of confocal microscopy in rheumatoid arthritis patients with concomitant dry eye disease. We examined 40 rheumatoid arthritis patients with variable severity and 44 volunteer age- and gender-matched healthy control subjects. We found that all examined parameters were significantly lower (p < 0.05) in rheumatoid arthritis patients as opposed to the control samples: namely, the number of fibers, the total length of the nerves, the number of branch points on the main fibers and the total nerve-fiber area. We examined further variables, such as age, sex and the duration of rheumatoid arthritis. Interestingly, we could not find a correlation between the above variables and abnormal neural structural changes in the cornea. We interpreted these findings via implementing our hypotheses. Correspondingly, one neuroimmunological link between dry eye and rheumatoid arthritis could be through the chronic Piezo2 channelopathy-induced K2P-TASK1 signaling axis. This could accelerate neuroimmune-induced sensitization on the spinal level in this autoimmune disease, with Langerhans-cell activation in the cornea and theorized downregulated Piezo1 channels in these cells. Even more importantly, suggested principal primary-damage-associated corneal keratocyte activation could be accompanied by upregulation of Piezo1. Both activation processes on the periphery would skew the plasticity of the Th17/Treg ratio, resulting in Th17/Treg imbalance in dry eye, secondary to rheumatoid arthritis. Hence, chronic somatosensory-terminal Piezo2 channelopathy-induced impaired Piezo2–Piezo1 crosstalk could result in a mixed picture of disrupted functional regeneration but upregulated morphological regeneration activity of these somatosensory axons in the cornea, providing the demonstrated abnormal neural corneal morphology.
Acute Cardiovascular and Cardiorespiratory Effects of JWH-018 in Awake and Freely Moving Mice: Mechanism of Action and Possible Antidotal Interventions?
Authors: Marchetti Beatrice , Bilel Sabrine , Tirri Micaela , Corli Giorgia , Roda Elisa , Locatelli Carlo Alessandro , Cavarretta Elena , De-Giorgio Fabio , Marti Matteo
Published: April 2023

JWH-018 is the most known compound among synthetic cannabinoids (SCs) used for their psychoactive effects. SCs-based products are responsible for several intoxications in humans. Cardiac toxicity is among the main side effects observed in emergency departments: SCs intake induces harmful effects such as hypertension, tachycardia, chest pain, arrhythmias, myocardial infarction, breathing impairment, and dyspnea. This study aims to investigate how cardio-respiratory and vascular JWH-018 (6 mg/kg) responses can be modulated by antidotes already in clinical use. The tested antidotes are amiodarone (5 mg/kg), atropine (5 mg/kg), nifedipine (1 mg/kg), and propranolol (2 mg/kg). The detection of heart rate, breath rate, arterial oxygen saturation (SpO2), and pulse distention are provided by a non-invasive apparatus (Mouse Ox Plus) in awake and freely moving CD-1 male mice. Tachyarrhythmia events are also evaluated. Results show that while all tested antidotes reduce tachycardia and tachyarrhythmic events and improve breathing functions, only atropine completely reverts the heart rate and pulse distension. These data may suggest that cardiorespiratory mechanisms of JWH-018-induced tachyarrhythmia involve sympathetic, cholinergic, and ion channel modulation. Current findings also provide valuable impetus to identify potential antidotal intervention to support physicians in the treatment of intoxicated patients in emergency clinical settings.
Osteosarcopenia in NAFLD/MAFLD: An Underappreciated Clinical Problem in Chronic Liver Disease
Authors: Musio Alessandra , Perazza Federica , Leoni Laura , Stefanini Bernardo , Dajti Elton , Menozzi Renata , Petroni Maria Letizia , Colecchia Antonio , Ravaioli Federico
Published: April 2023

Chronic liver disease (CLD), including non-alcoholic fatty liver disease (NAFLD) and its advanced form, non-alcoholic steatohepatitis (NASH), affects a significant portion of the population worldwide. NAFLD is characterised by fat accumulation in the liver, while NASH is associated with inflammation and liver damage. Osteosarcopenia, which combines muscle and bone mass loss, is an emerging clinical problem in chronic liver disease that is often underappreciated. The reductions in muscle and bone mass share several common pathophysiological pathways; insulin resistance and chronic systemic inflammation are the most crucial predisposing factors and are related to the presence and gravity of NAFLD and to the worsening of the outcome of liver disease. This article explores the relationship between osteosarcopenia and NAFLD/MAFLD, focusing on the diagnosis, prevention and treatment of this condition in patients with CLD.
Epigenetic Gene-Regulatory Loci in Alu Elements Associated with Autism Susceptibility in the Prefrontal Cortex of ASD
Authors: Saeliw Thanit , Kanlayaprasit Songphon , Thongkorn Surangrat , Songsritaya Kwanjira , Sanannam Bumpenporn , Sae-Lee Chanachai , Jindatip Depicha , Hu Valerie W. , Sarachana Tewarit
Published: April 2023

Alu elements are transposable elements that can influence gene regulation through several mechanisms; nevertheless, it remains unclear whether dysregulation of Alu elements contributes to the neuropathology of autism spectrum disorder (ASD). In this study, we characterized transposable element expression profiles and their sequence characteristics in the prefrontal cortex tissues of ASD and unaffected individuals using RNA-sequencing data. Our results showed that most of the differentially expressed transposable elements belong to the Alu family, with 659 loci of Alu elements corresponding to 456 differentially expressed genes in the prefrontal cortex of ASD individuals. We predicted cis- and trans-regulation of Alu elements to host/distant genes by conducting correlation analyses. The expression level of Alu elements correlated significantly with 133 host genes (cis-regulation, adjusted p < 0.05) associated with ASD as well as the cell survival and cell death of neuronal cells. Transcription factor binding sites in the promoter regions of differentially expressed Alu elements are conserved and associated with autism candidate genes, including RORA. COBRA analyses of postmortem brain tissues showed significant hypomethylation in global methylation analyses of Alu elements in ASD subphenotypes as well as DNA methylation of Alu elements located near the RNF-135 gene (p < 0.05). In addition, we found that neuronal cell density, which was significantly increased (p = 0.042), correlated with the expression of genes associated with Alu elements in the prefrontal cortex of ASD. Finally, we determined a relationship between these findings and the ASD severity (i.e., ADI-R scores) of individuals with ASD. Our findings provide a better understanding of the impact of Alu elements on gene regulation and molecular neuropathology in the brain tissues of ASD individuals, which deserves further investigation.
Comparative Genomic Analysis of a Methylorubrum rhodesianum MB200 Isolated from Biogas Digesters Provided New Insights into the Carbon Metabolism of Methylotrophic Bacteria
Authors: Zhang Xi , Xia Liqing , Liu Jianyi , Wang Zihao , Yang Yanni , Wu Yiting , Yang Qingshan , Huang Luodong , Shen Peihong
Published: April 2023

Methylotrophic bacteria are widely distributed in nature and can be applied in bioconversion because of their ability to use one-carbon source. The aim of this study was to investigate the mechanism underlying utilization of high methanol content and other carbon sources by Methylorubrum rhodesianum strain MB200 via comparative genomics and analysis of carbon metabolism pathway. The genomic analysis revealed that the strain MB200 had a genome size of 5.7 Mb and two plasmids. Its genome was presented and compared with that of the 25 fully sequenced strains of Methylobacterium genus. Comparative genomics revealed that the Methylorubrum strains had closer collinearity, more shared orthogroups, and more conservative MDH cluster. The transcriptome analysis of the strain MB200 in the presence of various carbon sources revealed that a battery of genes was involved in the methanol metabolism. These genes are involved in the following functions: carbon fixation, electron transfer chain, ATP energy release, and resistance to oxidation. Particularly, the central carbon metabolism pathway of the strain MB200 was reconstructed to reflect the possible reality of the carbon metabolism, including ethanol metabolism. Partial propionate metabolism involved in ethyl malonyl-CoA (EMC) pathway might help to relieve the restriction of the serine cycle. In addition, the glycine cleavage system (GCS) was observed to participate in the central carbon metabolism pathway. The study revealed the coordination of several metabolic pathways, where various carbon sources could induce associated metabolic pathways. To the best of our knowledge, this is the first study providing a more comprehensive understanding of the central carbon metabolism in Methylorubrum. This study provided a reference for potential synthetic and industrial applications of this genus and its use as chassis cells.
An Overview of Factors Affecting the Functional Quality of Common Wheat (Triticum aestivum L.)
Authors: Filip Ewa , Woronko Karolina , Stępień Edyta , Czarniecka Natalia
Published: April 2023

Wheat (Triticum aestivum L.) is one of the most important crops worldwide, and, as a resilient cereal, it grows in various climatic zones. Due to changing climatic conditions and naturally occurring environmental fluctuations, the priority problem in the cultivation of wheat is to improve the quality of the crop. Biotic and abiotic stressors are known factors leading to the deterioration of wheat grain quality and to crop yield reduction. The current state of knowledge on wheat genetics shows significant progress in the analysis of gluten, starch, and lipid genes responsible for the synthesis of the main nutrients in the endosperm of common wheat grain. By identifying these genes through transcriptomics, proteomics, and metabolomics studies, we influence the creation of high-quality wheat. In this review, previous works were assessed to investigate the significance of genes, puroindolines, starches, lipids, and the impact of environmental factors, as well as their effects on the wheat grain quality.
Multiple Nucleocapsid Structural Forms of Shrimp White Spot Syndrome Virus Suggests a Novel Viral Morphogenetic Pathway
Authors: Huang Hui-Ju , Tang Sen-Lin , Chang Yuan-Chih , Wang Hao-Ching , Ng Tze Hann , Garmann Rees F. , Chen Yu-Wen , Huang Jiun-Yan , Kumar Ramya , Chang Sheng-Hsiung , Wu Shang-Rung , Chao Chih-Yu , Matoba Kyoko , Kenji Iwasaki , Gelbart William M. , Ko Tzu-Ping , Wang Hwei-Jiung (Andrew) , Lo Chu-Fang , Chen Li-Li , Wang Han-Ching
Published: April 2023

White spot syndrome virus (WSSV) is a very large dsDNA virus. The accepted shape of the WSSV virion has been as ellipsoidal, with a tail-like extension. However, due to the scarcity of reliable references, the pathogenesis and morphogenesis of WSSV are not well understood. Here, we used transmission electron microscopy (TEM) and cryogenic electron microscopy (Cryo-EM) to address some knowledge gaps. We concluded that mature WSSV virions with a stout oval-like shape do not have tail-like extensions. Furthermore, there were two distinct ends in WSSV nucleocapsids: a portal cap and a closed base. A C14 symmetric structure of the WSSV nucleocapsid was also proposed, according to our Cryo-EM map. Immunoelectron microscopy (IEM) revealed that VP664 proteins, the main components of the 14 assembly units, form a ring-like architecture. Moreover, WSSV nucleocapsids were also observed to undergo unique helical dissociation. Based on these new results, we propose a novel morphogenetic pathway of WSSV.
Identification of the GDP-L-Galactose Phosphorylase Gene as a Candidate for the Regulation of Ascorbic Acid Content in Fruits of Capsicum annuum L.
Authors: Wang Yixin , Wang Zheng , Geng Sansheng , Du Heshan , Chen Bin , Sun Liang , Wang Guoyun , Sha Meihong , Dong Tingting , Zhang Xiaofen , Wang Qian
Published: April 2023

Ascorbic acid (AsA) is an antioxidant with significant functions in both plants and animals. Despite its importance, there has been limited research on the molecular basis of AsA production in the fruits of Capsicum annuum L. In this study, we used Illumina transcriptome sequencing (RNA-seq) technology to explore the candidate genes involved in AsA biosynthesis in Capsicum annuum L. A total of 8272 differentially expressed genes (DEGs) were identified by the comparative transcriptome analysis. Weighted gene co-expression network analysis identified two co-expressed modules related to the AsA content (purple and light-cyan modules), and eight interested DEGs related to AsA biosynthesis were selected according to gene annotations in the purple and light-cyan modules. Moreover, we found that the gene GDP-L-galactose phosphorylase (GGP) was related to AsA content, and silencing GGP led to a reduction in the AsA content in fruit. These results demonstrated that GGP is an important gene controlling AsA biosynthesis in the fruit of Capsicum annuum L. In addition, we developed capsanthin/capsorubin synthase as the reporter gene for visual analysis of gene function in mature fruit, enabling us to accurately select silenced tissues and analyze the results of silencing. The findings of this study provide the theoretical basis for future research to elucidate AsA biosynthesis in Capsicum annuum L.
Coronary Artery Spasm-Related Heart Failure Syndrome: Literature Review
Authors: Hung Ming-Jui , Yeh Chi-Tai , Kounis Nicholas G. , Koniari Ioanna , Hu Patrick , Hung Ming-Yow
Published: April 2023

Although heart failure (HF) is a clinical syndrome that becomes worse over time, certain cases can be reversed with appropriate treatments. While coronary artery spasm (CAS) is still underappreciated and may be misdiagnosed, ischemia due to coronary artery disease and CAS is becoming the single most frequent cause of HF worldwide. CAS could lead to syncope, HF, arrhythmias, and myocardial ischemic syndromes such as asymptomatic ischemia, rest and/or effort angina, myocardial infarction, and sudden death. Albeit the clinical significance of asymptomatic CAS has been undervalued, affected individuals compared with those with classic Heberden’s angina pectoris are at higher risk of syncope, life-threatening arrhythmias, and sudden death. As a result, a prompt diagnosis implements appropriate treatment strategies, which have significant life-changing consequences to prevent CAS-related complications, such as HF. Although an accurate diagnosis depends mainly on coronary angiography and provocative testing, clinical characteristics may help decision-making. Because the majority of CAS-related HF (CASHF) patients present with less severe phenotypes than overt HF, it underscores the importance of understanding risk factors correlated with CAS to prevent the future burden of HF. This narrative literature review summarises and discusses separately the epidemiology, clinical features, pathophysiology, and management of patients with CASHF.
Finding New Molecular Targets of Two Copper(II)-Hydrazone Complexes on Triple-Negative Breast Cancer Cells Using Mass-Spectrometry-Based Quantitative Proteomics
Authors: Balsa Lucia M. , Rodriguez María R. , Ferraresi-Curotto Verónica , Parajón-Costa Beatriz S. , Gonzalez-Baró Ana C. , León Ignacio E.
Published: April 2023

Breast cancer is the most common cancer in women, with a high incidence estimated to reach 2.3 million by 2030. Triple-Negative Breast Cancer (TNBC) is the greatest invasive class of breast cancer with a poor prognosis, due to the side-effects exerted by the chemotherapy used and the low effectivity of novel treatments. In this sense, copper compounds have shown to be potentially effective as antitumor agents, attracting increasing interest as alternatives to the usually employed platinum-derived drugs. Therefore, the aim of this work is to identify differentially expressed proteins in MDA-MB-231 cells exposed to two copper(II)-hydrazone complexes using label-free quantitative proteomics and functional bioinformatics strategies to identify the molecular mechanisms through which these copper complexes exert their antitumoral effect in TNBC cells. Both copper complexes increased proteins involved in endoplasmic reticulum stress and unfolded protein response, as well as the downregulation of proteins related to DNA replication and repair. One of the most relevant anticancer mechanisms of action found for CuHL1 and CuHL2 was the down-regulation of gain-of-function-mutant p53. Moreover, we found a novel and interesting effect for a copper metallodrug, which was the down-regulation of proteins related to lipid synthesis and metabolism that could lead to a beneficial decrease in lipid levels.
Differential Effects of Hypoxia versus Hyperoxia or Physoxia on Phenotype and Energy Metabolism in Human Chondrocytes from Osteoarthritic Compared to Macroscopically Normal Cartilage
Authors: Jain Lekha , Bolam Scott M. , Monk A. Paul , Munro Jacob T. , Chen Even , Tamatea Jade , Dalbeth Nicola , Poulsen Raewyn C.
Published: April 2023

Chondrocyte phenotype and energy metabolism are altered in osteoarthritis (OA). However, most studies characterising the change in human chondrocyte behaviour in OA have been conducted in supraphysiological oxygen concentrations. The purpose of this study was to compare phenotype and energy metabolism in chondrocytes from macroscopically normal (MN) and OA cartilage maintained in 18.9% (standard tissue culture), 6% (equivalent to superficial zone of cartilage in vivo) or 1% oxygen (equivalent to deep zone of cartilage in vivo). MMP13 production was higher in chondrocytes from OA compared to MN cartilage in hyperoxia and physoxia but not hypoxia. Hypoxia promoted SOX9, COL2A1 and ACAN protein expression in chondrocytes from MN but not OA cartilage. OA chondrocytes used higher levels of glycolysis regardless of oxygen availability. These results show that differences in phenotype and energy metabolism between chondrocytes from OA and MN cartilage differ depending on oxygen availability. OA chondrocytes show elevated synthesis of cartilage-catabolising enzymes and chondrocytes from MN cartilage show reduced cartilage anabolism in oxygenated conditions. This is relevant as a recent study has shown that oxygen levels are elevated in OA cartilage in vivo. Our findings may indicate that this elevated cartilage oxygenation may promote cartilage loss in OA.