bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and Characterization of Two Putative Citrus Phosphomannose Isomerase (CsPMI) Genes as Selectable Markers for Mature Citrus Transformation
Authors: Wu Hao , Canton Michel , Mahmoud Lamiaa M. , Weber Katherine R. , Michalczyk Gillian Z. , Dutt Manjul , Zale Janice M.
Published: February 2022

Two Citrus sinensis (L.) phosphomannose isomerase (PMI) genes, CsPMI1 and CsPMI2, were evaluated as novel selectable markers in mature citrus transformation. Transgenic shoots produced after transformation of Kuharske rootstock with each PMI construct were selected on six treatments of mannose and sucrose. For CsPMI1, there were no significant differences among the various mannose and sucrose treatments for the mean number of positive shoots (PS), the mean transformation efficiency based on the number of shoots (TES), or the mean transformation efficiency based on the number of explants (TEE). However, for the CsPMI2 gene, the number of transgenics produced in two treatments (7.5 g L−1 mannose + 22.5 g L−1 sucrose and 15 g L−1 mannose + 15 g L−1 sucrose) was significantly greater than the sucrose control for TES at 4.2% and 3.7%, respectively. Moreover, TEE at 4.2% in the 15 g L−1 mannose + 15 g L−1 sucrose treatment, supported the TES value. Most of the transgenic lines demonstrated higher in vivo and in vitro enzyme assays compared with the wild-type control. CsPMI2 provided acceptable selection in mature citrus, and it will be applied in future intragenic research.
Role of Reactive Oxygen Species against Pathogens in Relation to Postharvest Disease of Papaya Fruit
Authors: Feng Linyan , Sun Jian , Jiang Yueming , Duan Xuewu
Published: February 2022

Reactive oxygen species (ROS) play an active role in plant defense. Polyphenol oxidase (PPO) and peroxidase (POD) participate in the synthesis of phytoalexins. The comparative activities of ROS, including hydrogen peroxide (H2O2), superoxide anions (O2−), and hydroxyl radicals (·OH), against the fungal pathogen Colletotrichum gloeosporioides from papaya fruit were evaluated. The effects of ROS on PPO and POD activities in papaya fruit inoculated with C. gloeosporioides and the development of natural decay in intact fruit were also investigated. ·OH was the most effective in inhibiting conidial germination and mycelial growth of C. gloeosporioides in vitro. However, 20 or 30 mM H2O2 exhibited the best control of the three ROS treatments at ameliorating the disease symptoms associated with the highest levels of PPO and POD activities in papaya fruit. Furthermore, the activities of PPO and POD negatively correlated with the disease index. Overall, H2O2 treatments can induce the resistance of papaya fruit against C. gloeosporioides owing to the enhanced activities of PPO and POD. Treatment with 20 mM H2O2 also significantly reduced the incidence of decay in intact papaya fruit in semi-commercial experiment, which could make it a potential alternative manner to control postharvest disease in papaya fruit.
Evaluation of the Different Low-Tech Protective Cultivation Approaches to Improve Yield and Phytochemical Accumulation of Papaya (Carica papaya L.) in Bangladesh
Authors: Choudhury Shormin , Islam Nazrul , Mustaki Sika , Uddain Jasim , Azad Md Obyedul Kalam , Choi Ki Young , Naznin Most Tahera
Published: February 2022

The production of horticultural crops in the outdoor environment facing various environmental factors, such as cyclones, droughts, heavy rain, and hailstorms, significantly affects the papaya production in the sub-tropical regions, especially in Bangladesh. Protected cultivation of horticultural crops is a common practice in developed countries. However, it is rarely observed in the developing countries, particularly for papaya production. Therefore, this study was carried out to find a protective approach to papaya cultivation to mitigate the environmental factors to obtain a quality yield. This production system consists of three treatments, including net house, poly shed house, UV poly shed house, and open field conditions (control). The results revealed that plants grown in the net house had significantly higher leaf number (30), fruit number (68), and fruit yield (56.28 kg/plant) than the control grown plant. Papaya cultured in the net house also showed significantly higher accumulation of chlorophyll, ascorbic acid, total phenol, reducing sugar, and β-carotene than those grown in other environments. In terms of peel color, papaya grown in the net house had the highest a* value (redness), whereas that grown in the open field had the lowest. Thus, the study demonstrated that papaya can be cultivated successfully in a net house with increased yield and phytochemical content. The findings provide a fundamental production strategy for quality papaya production in Bangladesh.
Measuring Equine-Assisted Therapy: Validation and Confirmatory Factor Analysis of an ICF-Based Standardized Assessment-Tool
Authors: Stolz Isabel , Anneken Volker , Froböse Ingo
Published: February 2022

The International Classification of Functioning, Disability, and Health (ICF) of the World Health Organization (WHO) was established as an international framework for monitoring rehabilitation outcomes and the impacts of health interventions since, as the term “functioning” implies, it emphasizes a person’s “lived health” in addition to their biological health status. Equine-assisted therapy (EAT) represents a holistic intervention approach that aims to improve both biomedical functioning and the patient’s lived health in relation to performing activities and participating in social situations. In this study, the psychometric properties of an ICF-based digital assessment tool for the measurement of the rehabilitation impacts of EAT were analyzed via simultaneous confirmatory factor analyses (CFA) and reliability and sensitivity tests. In total, 265 patients from equine-assisted therapy centers in Germany were included for CFA. Change sensitivity was assessed via multi-level analyses based on 876 repeated assessments by 30 therapists. Results show satisfactory model-fit statistics; McDonald’s omega (ML) showed excellent scores for the total scale (ω = 0.96) and three subscales (ω = 0.95; ω = 0.95, ω = 0.93). The tool proved itself to be change sensitive and reliable (change sensitivity p ≤ 0.001), retest r = 0.745 **, p ≤ 0.001). Overall, the developed assessment tool satisfactorily fulfills psychometric requirements and can be applied in therapeutic practice.
Biocompatible Materials in Otorhinolaryngology and Their Antibacterial Properties
Authors: Spałek Jakub , Ociepa Przemysław , Deptuła Piotr , Piktel Ewelina , Daniluk Tamara , Król Grzegorz , Góźdź Stanisław , Bucki Robert , Okła Sławomir
Published: February 2022

For decades, biomaterials have been commonly used in medicine for the replacement of human body tissue, precise drug-delivery systems, or as parts of medical devices that are essential for some treatment methods. Due to rapid progress in the field of new materials, updates on the state of knowledge about biomaterials are frequently needed. This article describes the clinical application of different types of biomaterials in the field of otorhinolaryngology, i.e., head and neck surgery, focusing on their antimicrobial properties. The variety of their applications includes cochlear implants, middle ear prostheses, voice prostheses, materials for osteosynthesis, and nasal packing after nasal/paranasal sinuses surgery. Ceramics, such as as hydroxyapatite, zirconia, or metals and metal alloys, still have applications in the head and neck region. Tissue engineering scaffolds and drug-eluting materials, such as polymers and polymer-based composites, are becoming more common. The restoration of life tissue and the ability to prevent microbial colonization should be taken into consideration when designing the materials to be used for implant production. The authors of this paper have reviewed publications available in PubMed from the last five years about the recent progress in this topic but also establish the state of knowledge of the most common application of biomaterials over the last few decades.
Biomarkers in Human Peripheral Blood Mononuclear Cells: The State of the Art in Amyotrophic Lateral Sclerosis
Authors: Pansarasa Orietta , Garofalo Maria , Scarian Eveljn , Dragoni Francesca , Garau Jessica , Di Gerlando Rosalinda , Diamanti Luca , Bordoni Matteo , Gagliardi Stella
Published: February 2022

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease, characterized by the progressive loss of lower motor neurons, weakness and muscle atrophy. ALS lacks an effective cure and diagnosis is often made by exclusion. Thus, it is imperative to search for biomarkers. Biomarkers can help in understanding ALS pathomechanisms, identification of targets for treatment and development of effective therapies. Peripheral blood mononuclear cells (PBMCs) represent a valid source for biomarkers compared to cerebrospinal fluid, as they are simple to collect, and to plasma, because of the possibility of detecting lower expressed proteins. They are a reliable model for patients’ stratification. This review provides an overview on PBMCs as a potential source of biomarkers in ALS. We focused on altered RNA metabolism (coding/non-coding RNA), including RNA processing, mRNA stabilization, transport and translation regulation. We addressed protein abnormalities (aggregation, misfolding and modifications); specifically, we highlighted that SOD1 appears to be the most characterizing protein in ALS. Finally, we emphasized the correlation between biological parameters and disease phenotypes, as regards prognosis, severity and clinical features. In conclusion, even though further studies are needed to standardize the use of PBMCs as a tool for biomarker investigation, they represent a promising approach in ALS research.
The Finely Coordinated Action of SSB and NurA/HerA Complex Strictly Regulates the DNA End Resection Process in Saccharolobus solfataricus
Authors: De Falco Mariarosaria , Porritiello Alessandra , Rota Federica , Scognamiglio Viviana , Antonacci Amina , del Monaco Giovanni , De Felice Mariarita
Published: February 2022

Generation of the 3′ overhang is a critical step during homologous recombination (HR) and replication fork rescue processes. This event is usually performed by a series of DNA nucleases and/or helicases. The nuclease NurA and the ATPase HerA, together with the highly conserved MRE11/RAD50 proteins, play an important role in generating 3′ single-stranded DNA during archaeal HR. Little is known, however, about HerA-NurA function and activation of this fundamental and complicated DNA repair process. Herein, we analyze the functional relationship among NurA, HerA and the single-strand binding protein SSB from Saccharolubus solfataricus. We demonstrate that SSB clearly inhibits NurA endonuclease activity and its exonuclease activities also when in combination with HerA. Moreover, we show that SSB binding to DNA is greatly stimulated by the presence of either NurA or NurA/HerA. In addition, if on the one hand NurA binding is not influenced, on the other hand, HerA binding is reduced when SSB is present in the reaction. In accordance with what has been observed, we have shown that HerA helicase activity is not stimulated by SSB. These data suggest that, in archaea, the DNA end resection process is governed by the strictly combined action of NurA, HerA and SSB.
Evaluating the Sub-Acute Toxicity of Formaldehyde Fumes in an In Vitro Human Airway Epithelial Tissue Model
Authors: Ren Baiping , Wu Qiangen , Muskhelishvili Levan , Davis Kelly , Wang Yiying , Rua Diego , Cao Xuefei
Published: February 2022

Formaldehyde (FA) is an irritating, highly reactive aldehyde that is widely regarded as an asthmagen. In addition to its use in industrial applications and being a product of combustion reaction and endogenous metabolism, FDA-regulated products may contain FA or release FA fumes that present toxicity risks for both patients and healthcare workers. Exposure to airborne FA is associated with nasal neoplastic lesions in both animals and humans. It is classified as a Group 1 carcinogen by International Agency for Research on Cancer (IARC) based on the increased incidence of cancer in animals and a known human carcinogen in the Report on Carcinogens by National Toxicology Program (NTP). Herein, we systematically evaluated the tissue responses to FA fumes in an in vitro human air-liquid-interface (ALI) airway tissue model. Cultures were exposed at the air interface to 7.5, 15, and 30 ppm of FA fumes 4 h per day for 5 consecutive days. Exposure to 30 ppm of FA induced sustained oxidative stress, along with functional changes in ciliated and goblet cells as well as possible squamous differentiation. Furthermore, secretion of the proinflammatory cytokines, IL-1β, IL-2, IL-8, GM-CSF, TNF-a and IFN-γ, was induced by repeated exposures to FA fumes. Expression of MMP-1, MMP-3, MMP-7, MMP-10, MMP-12, and MMP-13 was downregulated at the end of the 5-day exposure. Although DNA-damage was not detected by the comet assay, FA exposures downregulated the DNA repair enzymes MGMT and FANCD2, suggesting its possible interference in the DNA repair capacity. Overall, a general concordance was observed between our in vitro responses to FA fume exposures and the reported in vivo toxicity of FA. Our findings provide further evidence supporting the application of the ALI airway system as a potential in vitro alternative for screening and evaluating the respiratory toxicity of inhaled substances.
Amelioration of Endotoxin-Induced Acute Lung Injury and Alveolar Epithelial Cells Apoptosis by Simvastatin Is Associated with Up-Regulation of Survivin/NF-kB/p65 Pathway
Authors: Nežić Lana , Amidžić Ljiljana , Škrbić Ranko , Gajanin Radoslav , Mandić Danijela , Dumanović Jelena , Milovanović Zoran , Jaćević Vesna
Published: February 2022

Disruption of the alveolar–endothelial barrier caused by inflammation leads to the progression of septic acute lung injury (ALI). In the present study, we investigated the beneficial effects of simvastatin on the endotoxin lipopolysaccharide (LPS)-induced ALI and its related mechanisms. A model of ALI was induced within experimental sepsis developed by intraperitoneal injection of a single non-lethal LPS dose after short-term simvastatin pretreatment (10–40 mg/kg orally). The severity of the lung tissue inflammatory injury was expressed as pulmonary damage scores (PDS). Alveolar epithelial cell apoptosis was confirmed by TUNEL assay (DNA fragmentation) and expressed as an apoptotic index (AI), and immunohistochemically for cleaved caspase-3, cytochrome C, and anti-apoptotic Bcl-xL, an inhibitor of apoptosis, survivin, and transcriptional factor, NF-kB/p65. Severe inflammatory injury of pulmonary parenchyma (PDS 3.33 ± 0.48) was developed after the LPS challenge, whereas simvastatin significantly and dose-dependently protected lung histology after LPS (p < 0.01). Simvastatin in a dose of 40 mg/kg showed the most significant effects in amelioration alveolar epithelial cells apoptosis, demonstrating this as a marked decrease of AI (p < 0.01 vs. LPS), cytochrome C, and cleaved caspase-3 expression. Furthermore, simvastatin significantly enhanced the expression of Bcl-xL and survivin. Finally, the expression of survivin and its regulator NF-kB/p65 in the alveolar epithelium was in strong positive correlation across the groups. Simvastatin could play a protective role against LPS-induced ALI and apoptosis of the alveolar–endothelial barrier. Taken together, these effects were seemingly mediated by inhibition of caspase 3 and cytochrome C, a finding that might be associated with the up-regulation of cell-survival survivin/NF-kB/p65 pathway and Bcl-xL.
Renal Oncocytoma: The Diagnostic Challenge to Unmask the Double of Renal Cancer
Authors: Trevisani Francesco , Floris Matteo , Minnei Roberto , Cinque Alessandra
Published: February 2022

Renal oncocytoma represents the most common type of benign neoplasm that is an increasing concern for urologists, oncologists, and nephrologists due to its difficult differential diagnosis and frequent overtreatment. It displays a variable neoplastic parenchymal and stromal architecture, and the defining cellular element is a large polygonal, granular, eosinophilic, mitochondria-rich cell known as an oncocyte. The real challenge in the oncocytoma treatment algorithm is related to the misdiagnosis due to its resemblance, at an initial radiological assessment, to malignant renal cancers with a completely different prognosis and medical treatment. Unfortunately, percutaneous renal biopsy is not frequently performed due to the possible side effects related to the procedure. Therefore, the majority of oncocytoma are diagnosed after the surgical operation via partial or radical nephrectomy. For this reason, new reliable strategies to solve this issue are needed. In our review, we will discuss the clinical implications of renal oncocytoma in daily clinical practice with a particular focus on the medical diagnosis and treatment and on the potential of novel promising molecular biomarkers such as circulating microRNAs to distinguish between a benign and a malignant lesion.
Thiolated Hydroxypropyl-β-cyclodextrin: A Potential Multifunctional Excipient for Ocular Drug Delivery
Authors: Grassiri Brunella , Knoll Patrick , Fabiano Angela , Piras Anna Maria , Zambito Ylenia , Bernkop-Schnürch Andreas
Published: February 2022

The goal of this study was the design and evaluation of a thiolated cyclodextrin providing high drug solubilizing and mucoadhesive properties for ocular drug delivery. Hydroxypropyl-β-cyclodextrin (HP-β-CD) was thiolated via a microwave-assisted method, resulting in a degree of thiolation of 33%. Mucoadhesive properties of thiolated HP-β-CD (HP-β-CD-SH) were determined via rheological measurements and ex vivo studies on isolated porcine cornea. Due to thiolation of HP-β-CD, a 2-fold increase of mucus viscosity and a 1.4-fold increase in residence time on isolated corneal tissue were achieved. After instillation, the mean precorneal residence time and AUC of dexamethasone (DMS) eye drops were 4-fold and 11.7-fold enhanced by HP-β-CD-SH, respectively. Furthermore, in the presence of HP-β-CD-SH, a constant high level of DMS in aqueous humour between 30 and 150 min after administration was observed. These results suggest that HP-β-CD-SH is an excellent excipient for ocular formulations of poorly soluble drugs in order to prolong their ocular residence time and bioavailability.
Expression and Transcript Localization of star, sf-1, and dax-1 in the Early Brain of the Orange-Spotted Grouper Epinephelus coioides
Authors: Nagarajan Ganesan , Aruna Adimoolam , Alkhamis Yousef Ahmed , Mathew Roshmon Thomas , Chang Ching-Fong
Published: February 2022

We investigated the developmental expression and localization of sf-1 and dax-1 transcripts in the brain of the juvenile orange-spotted grouper in response to steroidogenic enzyme gene at various developmental ages in relation to gonadal sex differentiation. The sf-1 transcripts were significantly higher from 110-dah (day after hatching) and gradually increased up to 150-dah. The dax-1 mRNA, on the other hand, showed a decreased expression during this period, in contrast to sf-1 expression. At the same time, the early brain had increased levels of steroidogenic gene (star). sf-1 and star hybridization signals were found to be increased in the ventromedial hypothalamus at 110-dah; however, dax-1 mRNA signals decreased in the early brain toward 150-dah. Furthermore, the exogenous estradiol upregulated star and sf-1 transcripts in the early brain of the grouper. These findings suggest that sf-1 and dax-1 may have an antagonistic expression pattern in the early brain during gonadal sex differentiation. Increased expression of steroidogenic gene together with sf-1 during gonadal differentiation strongly suggests that sf-1 may play an important role in the juvenile grouper brain steroidogenesis and brain development.
The BDNF/TrkB Neurotrophin System in the Sensory Organs of Zebrafish
Authors: Aragona Marialuisa , Porcino Caterina , Guerrera Maria Cristina , Montalbano Giuseppe , Laurà Rosaria , Cometa Marzio , Levanti Maria , Abbate Francesco , Cobo Teresa , Capitelli Gabriel , Vega José A. , Germanà Antonino
Published: February 2022

The brain-derived neurotrophic factor (BDNF) was discovered in the last century, and identified as a member of the neurotrophin family. BDNF shares approximately 50% of its amino acid with other neurotrophins such as NGF, NT-3 and NT-4/5, and its linear amino acid sequences in zebrafish (Danio rerio) and human are 91% identical. BDNF functions can be mediated by two categories of receptors: p75NTR and Trk. Intriguingly, BDNF receptors were highly conserved in the process of evolution, as were the other NTs’ receptors. In this review, we update current knowledge about the distribution and functions of the BDNF-TrkB system in the sensory organs of zebrafish. In fish, particularly in zebrafish, the distribution and functions of BDNF and TrkB in the brain have been widely studied. Both components of the system, associated or segregated, are also present outside the central nervous system, especially in sensory organs including the inner ear, lateral line system, retina, taste buds and olfactory epithelium.
Properties and Mechanisms of Flavin-Dependent Monooxygenases and Their Applications in Natural Product Synthesis
Authors: Deng Yaming , Zhou Quan , Wu Yuzhou , Chen Xi , Zhong Fangrui
Published: February 2022

Natural products are usually highly complicated organic molecules with special scaffolds, and they are an important resource in medicine. Natural products with complicated structures are produced by enzymes, and this is still a challenging research field, its mechanisms requiring detailed methods for elucidation. Flavin adenine dinucleotide (FAD)-dependent monooxygenases (FMOs) catalyze many oxidation reactions with chemo-, regio-, and stereo-selectivity, and they are involved in the synthesis of many natural products. In this review, we introduce the mechanisms for different FMOs, with the classical FAD (C4a)-hydroperoxide as the major oxidant. We also summarize the difference between FMOs and cytochrome P450 (CYP450) monooxygenases emphasizing the advantages of FMOs and their specificity for substrates. Finally, we present examples of FMO-catalyzed synthesis of natural products. Based on these explanations, this review will expand our knowledge of FMOs as powerful enzymes, as well as implementation of the FMOs as effective tools for biosynthesis.
Modulatory Effects of Fractalkine on Inflammatory Response and Iron Metabolism of Lipopolysaccharide and Lipoteichoic Acid-Activated THP-1 Macrophages
Authors: Pandur Edina , Tamási Kitti , Pap Ramóna , Jánosa Gergely , Sipos Katalin
Published: February 2022

Fractalkine (CX3CL1) acts as a chemokine as well as a regulator of iron metabolism. Fractalkine binds CX3CR1, the fractalkine receptor on the surface of monocytes/macrophages regulating different intracellular signalling pathways such as mitogen-activated protein kinase (MAPK), phospholipase C (PLC) and NFκB contributing to the production of pro-inflammatory cytokine synthesis, and the regulation of cell growth, differentiation, proliferation and metabolism. In this study, we focused on the modulatory effects of fractalkine on the immune response and on the iron metabolism of Escherichia coli and Pseudomonas aeruginosa lipopolysaccharides (LPS) and Staphylococcus aureus lipoteichoic acid (LTA) activated THP-1 cells to get a deeper insight into the role of soluble fractalkine in the regulation of the innate immune system. Pro-inflammatory cytokine secretions of the fractalkine-treated, LPS/LTA-treated, and co-treated THP-1 cells were determined using ELISArray and ELISA measurements. We analysed the protein expression levels of signalling molecules regulated by CX3CR1 as well as hepcidin, the major iron regulatory hormone, the iron transporters, the iron storage proteins and mitochondrial iron utilization. The results showed that fractalkine treatment alone did not affect the pro-inflammatory cytokine secretion, but it was proposed to act as a regulator of the iron metabolism of THP-1 cells. In the case of two different LPS and one type of LTA with fractalkine co-treatments, fractalkine was able to alter the levels of signalling proteins (NFκB, PSTAT3, Nrf2/Keap-1) regulating the expression of pro-inflammatory cytokines as well as hepcidin, and the iron storage and utilization of the THP-1 cells.
Experimentally Induced Hyperinsulinemia Fails to Induce Polycystic Ovary Syndrome-like Traits in Female Rhesus Macaques
Authors: Zhou Rao , Bruns Cristin M. , Bird Ian M. , Kemnitz Joseph W. , Dumesic Daniel A. , Abbott David H.
Published: February 2022

As in women with polycystic ovary syndrome (PCOS), hyperinsulinemia is associated with anovulation in PCOS-like female rhesus monkeys. Insulin sensitizers ameliorate hyperinsulinemia and stimulate ovulatory menstrual cycles in PCOS-like monkeys. To determine whether hyperinsulinemia (>694 pmol/L), alone, induces PCOS-like traits, five PCOS-like female rhesus monkeys with minimal PCOS-like traits, and four control females of similar mid-to-late reproductive years and body mass index, received daily subcutaneous injections of recombinant human insulin or diluent for 6–7 months. A cross-over experimental design enabled use of the same monkeys in each treatment phase. Insulin treatment unexpectedly normalized follicular phase duration in PCOS-like, but not control, females. In response to an intramuscular injection of 200 IU hCG, neither prenatally androgenized nor control females demonstrated ovarian hyperandrogenic responses while receiving insulin. An intravenous GnRH (100 ng/kg) injection also did not reveal evidence of hypergonadotropism. Taken together, these results suggest that experimentally induced adult hyperinsulinemia, alone, is insufficient to induce PCOS-like traits in female rhesus monkeys and to amplify intrinsic PCOS-like pathophysiology.
Antifreeze Proteins: Novel Applications and Navigation towards Their Clinical Application in Cryobanking
Authors: Ekpo Marlene Davis , Xie Jingxian , Hu Yuying , Liu Xiangjian , Liu Fenglin , Xiang Jia , Zhao Rui , Wang Bo , Tan Songwen
Published: February 2022

Antifreeze proteins (AFPs) or thermal hysteresis (TH) proteins are biomolecular gifts of nature to sustain life in extremely cold environments. This family of peptides, glycopeptides and proteins produced by diverse organisms including bacteria, yeast, insects and fish act by non-colligatively depressing the freezing temperature of the water below its melting point in a process termed thermal hysteresis which is then responsible for ice crystal equilibrium and inhibition of ice recrystallisation; the major cause of cell dehydration, membrane rupture and subsequent cryodamage. Scientists on the other hand have been exploring various substances as cryoprotectants. Some of the cryoprotectants in use include trehalose, dimethyl sulfoxide (DMSO), ethylene glycol (EG), sucrose, propylene glycol (PG) and glycerol but their extensive application is limited mostly by toxicity, thus fueling the quest for better cryoprotectants. Hence, extracting or synthesizing antifreeze protein and testing their cryoprotective activity has become a popular topic among researchers. Research concerning AFPs encompasses lots of effort ranging from understanding their sources and mechanism of action, extraction and purification/synthesis to structural elucidation with the aim of achieving better outcomes in cryopreservation. This review explores the potential clinical application of AFPs in the cryopreservation of different cells, tissues and organs. Here, we discuss novel approaches, identify research gaps and propose future research directions in the application of AFPs based on recent studies with the aim of achieving successful clinical and commercial use of AFPs in the future.
An Anthranilate Derivative Inhibits Glutamate Release and Glutamate Excitotoxicity in Rats
Authors: Lu Cheng-Wei , Lin Chen-Jung , Hsieh Pei-Wen , Chiu Kuan-Ming , Lee Ming-Yi , Lin Tzu-Yu , Wang Su-Jane
Published: February 2022

The neurotransmitter glutamate plays an essential role in excitatory neurotransmission; however, excessive amounts of glutamate lead to excitotoxicity, which is the most common pathogenic feature of numerous brain disorders. This study aimed to investigate the role of butyl 2-[2-(2-fluorophenyl)acetamido]benzoate (HFP034), a synthesized anthranilate derivative, in the central glutamatergic system. We used rat cerebro-cortical synaptosomes to examine the effect of HFP034 on glutamate release. In addition, we used a rat model of kainic acid (KA)-induced glutamate excitotoxicity to evaluate the neuroprotective potential of HFP034. We showed that HFP034 inhibits 4-aminopyridine (4-AP)-induced glutamate release from synaptosomes, and this inhibition was absent in the absence of extracellular calcium. HFP034-mediated inhibition of glutamate release was associated with decreased 4-AP-evoked Ca2+ level elevation and had no effect on synaptosomal membrane potential. The inhibitory effect of HFP034 on evoked glutamate release was suppressed by blocking P/Q-type Ca2+ channels and protein kinase C (PKC). Furthermore, HFP034 inhibited the phosphorylation of PKC and its substrate, myristoylated alanine-rich C kinase substrate (MARCKS) in synaptosomes. We also observed that HFP034 pretreatment reduced neuronal death, glutamate concentration, glial activation, and the levels of endoplasmic reticulum stress-related proteins, calpains, glucose-regulated protein 78 (GRP 78), C/EBP homologous protein (CHOP), and caspase-12 in the hippocampus of KA-injected rats. We conclude that HFP034 is a neuroprotective agent that prevents glutamate excitotoxicity, and we suggest that this effect involves inhibition of presynaptic glutamate release through the suppression of P/Q-type Ca2+ channels and PKC/MARCKS pathways.
Mitochondrial De Novo Assembly of Iron–Sulfur Clusters in Mammals: Complex Matters in a Complex That Matters
Authors: Perfitt Tyler L. , Martelli Alain
Published: February 2022

Iron–sulfur clusters (Fe–S or ISC) are essential cofactors that function in a wide range of biological pathways. In mammalian cells, Fe–S biosynthesis primarily relies on mitochondria and involves a concerted group of evolutionary-conserved proteins forming the ISC pathway. In the early stage of the ISC pathway, the Fe–S core complex is required for de novo assembly of Fe–S. In humans, the Fe–S core complex comprises the cysteine desulfurase NFS1, the scaffold protein ISCU2, frataxin (FXN), the ferredoxin FDX2, and regulatory/accessory proteins ISD11 and Acyl Carrier Protein (ACP). In recent years, the field has made significant advances in unraveling the structure of the Fe–S core complex and the mechanism underlying its function. Herein, we review the key recent findings related to the Fe–S core complex and its components. We highlight some of the unanswered questions and provide a model of the Fe–S assembly within the complex. In addition, we briefly touch on the genetic diseases associated with mutations in the Fe–S core complex components.
Coronary Artery Disease and Atrial Fibrillation: A Bidirectional Mendelian Randomization Study
Authors: Yan Tao , Zhu Shijie , Xie Changming , Zhu Miao , Weng Fan , Wang Chunsheng , Guo Changfa
Published: February 2022

Background: Several works of observational clinical research indicate that coronary artery disease (CAD) and atrial fibrillation (AF) aggravate each other. However, it is unknown whether these associations reveal independent causal processes. Objective: The present study aimed to evaluate causal associations between CAD and AF using two-sample Mendelian randomization (TSMR) analysis. Methods: Summary-level Genome-wide association study (GWAS) data for CAD were obtained from the CARDIoGRAMplusC4D consortium, including 60,801 patients and 123,504 controls. General data for AF were acquired from the largest meta-analysis, comprising of 60,620 patients with AF and 970,216 non-cases. After data harmonization, three different methods—inverse-variance weighted (IVW), MR-Egger, and weighted-median—were applied for TSMR analysis. Results: The calculated ORs (95% CIs) for AF using IVW, MR-Egger, and weighted-median analysis were 1.11 (1.05, 1.17; p-value < 0.001), 1.14 (1.00, 1.29; p-value = 0.049), and 1.13 (1.08, 1.19; p-value < 0.001), respectively; for CAD, the results were 1.01 (0.97, 1.04; p-value = 0.76), 0.95 (0.89, 1.02; p-value = 0.15), and 1.00 (0.95, 1.05; p-value = 0.97). Conclusion: This comprehensive TSMR analysis provides evidence that patients with CAD are associated with an increased risk of AF. However, no causal association was found between patients with AF and the risk of CAD. These findings benefit clinical decision-making. Early heart-rhythm monitoring should be performed in patients with CAD. The prevention and treatment of AF complications such as thrombosis may be essential to reduce the incidence of CAD in AF patients.
The Safety of Anti-SARS-CoV-2 Vaccines: Vigilance Is Still Required
Authors: Goldman Michel
Published: February 2022

The opinion I put forward in this paper is that attention must continue to be paid to clinical observations compatible with a detrimental effect of anti-SARS-CoV-2 in certain diseases of immunological nature. Using the example of the atypical thrombocytopenic thromboses caused by adenoviral-vector-based vaccines, I argue that usual post-marketing pharmacovigilance programs may fail in identifying very rare vaccine-related disorders. Since the robust protective immunity induced by mRNA vaccines is related to their distinct capacity to induce strong stimulation of T follicular helper cells, I suggest that the safety of mRNA vaccines should be further assessed by appropriately designed epidemiological and mechanistic studies focusing on lymphoproliferative and autoimmune diseases in which T follicular helper cells were found to play a key role.
Frozen Section of Parotid Gland Tumours: The Head and Neck Pathologist as a Key Member of the Surgical Team
Authors: Mantsopoulos Konstantinos , Bessas Zacharias , Sievert Matti , Müller Sarina , Koch Michael , Agaimy Abbas , Iro Heinrich
Published: February 2022

Introduction: The aim of this study was to evaluate the impact of subspecialised head and neck versus general surgical pathologists on the reliability of the histopathologic evaluation during intraoperative consultation. Materials and Methods: The medical records of all patients who underwent a parotidectomy with frozen section between 2006 and 2021 were retrospectively evaluated. The frozen section was evaluated for sensitivity, specificity, accuracy, and predictive value. Assessment by two groups of pathologists (subspecialised head and neck versus general surgical pathologists) was compared, and the nature or types of misdiagnoses compared with final diagnoses on paraffin sections were analysed for the two groups. Results: Our study sample was made up of 669 cases. The mean age of patients was 57.7 years (range: 10–94 years). Of these, 490 patients had a benign lesion (73.2%), whereas 179 patients had a malignant lesion (26.8%). Frozen section had an overall accuracy of 97.6%, sensitivity for malignancy was 91.1%, specificity was 100%, PPV was 100%, and the NPV was 96.8%. The exact histologic subtype in the group of malignant tumours was correctly identified in FS in 89.4% of cases. A comparison of head and neck pathologists versus general surgical pathologists revealed a highly statistically significant difference concerning both overall detection of malignancy (p < 0.001) as well as correct identification of the histologic subtype (p < 0.001). Conclusion: Involvement of subspecialised head and neck pathologists in the intraoperative consultation for salivary gland tumours results in a gain of 19.8% more sensitivity, underlining the importance of specialisation in salivary gland pathology for the optimisation of frozen section quality.
Behavioral Activation through Virtual Reality for Depression: A Single Case Experimental Design with Multiple Baselines
Authors: Colombo Desirée , Suso-Ribera Carlos , Ortigosa-Beltrán Isabel , Fernández-Álvarez Javier , García-Palacios Azucena , Botella Cristina
Published: February 2022

Behavioral activation (BA) is a structured psychotherapeutic approach for the treatment of major depressive disorder (MDD), which aims at increasing the engagement in activities that might bring enjoyment and meaning to patients’ lives. Although a growing body of evidence supports the effectiveness of BA, enhancing the motivation and activity level of depressed patients is often challenging. In the present study, we explored the effectiveness of a brief BA treatment supported by virtual reality (VR) to facilitate the visualization and anticipation of four pleasurable activities that we tried to re-introduce in the patients’ daily routine. To do so, we conducted a single-case experimental design with multiple baselines in a sample of patients with moderate to severe depressive symptoms. Three overlap analyses across participants and across behaviors were conducted to calculate the rate of improvement of each patient after the delivery of the intervention. Across the three overlap indices, the participants generally showed moderate-to-large improvements in the level of daily activity, as well as in the time spent planning and/or engaging in one or more activities scheduled during the intervention. Furthermore, most patients also reported a moderate-to-large reduction in daily depressive symptoms and improved mood. Overall, the promising results of the present study suggest that the proposed VR-based BA intervention might represent a valid approach to behaviorally activate depressed patients. The barriers and future lines of research of this innovative field are discussed.
Free Insulin-like Growth Factor (IGF)-I in Children with PWS
Authors: Damen Layla , Elizabeth Melitza S. M. , Donze Stephany H. , van den Berg Sjoerd A. A. , de Graaff Laura C. G. , Hokken-Koelega Anita C. S.
Published: February 2022

In children with Prader–Willi syndrome (PWS), the standard growth hormone (GH) dose often results in high immunoreactive IGF-I levels. These high immunoreactive IGF-I levels lead to concern because their long-term effects are unknown. As a result, clinicians have to lower the GH dose, which worsens body composition and quality of life. As clinical features do not seem to correspond to immunoreactive IGF-I values, it is questionable whether immunoreactive IGF-I is a suitable marker for GH dosing, or whether another parameter better reflects IGF-I bioavailability and bioactivity. We, therefore, investigate serum immunoreactive IGF-I, free IGF-I and IGFBP-3 levels in 70 GH-treated children with PWS. Our study showed that, although immunoreactive IGF-I levels were high (>2 SDS) in the vast majority of prepubertal and pubertal children, free IGF-I SDS levels were <0 SDS in most and <1 SDS in all. Free IGF-I correlated with the immunoreactive IGF-I, IGFBP-3 and IGF-I/IGFBP-3 ratio. We conclude that there is a major discrepancy between immunoreactive and free IGF-I levels. While in the majority of GH-treated children with PWS, immunoreactive IGF-I levels were high, free IGF-I levels were <0 SDS in most. Our data appear to be very reassuring and suggest that free IGF-I levels should also be taken into consideration when the immunoreactive IGF-I levels are >2 SDS in GH-treated children with PWS.
Developing a Mechanistic Approach to Sudden Death Prevention in Mitral Valve Prolapse
Authors: Kelley Brian P. , Chaudry Abdul Mateen , Syed Faisal F.
Published: February 2022

Sudden cardiac death (SCD) from ventricular fibrillation (VF) can occur in mitral valve prolapse (MVP) in the absence of other comorbidities including mitral regurgitation, heart failure or coronary disease. Although only a small proportion with MVP are at risk, it can affect young, otherwise healthy adults, most commonly premenopausal women, often as the first presentation of MVP. In this review, we discuss arrhythmic mechanisms in MVP and mechanistic approaches for sudden death risk assessment and prevention. We define arrhythmogenic or arrhythmic MVP (AMVP) as MVP associated with complex and frequent ventricular ectopy, and malignant MVP (MMVP) as MVP with high risk of SCD. Factors predisposing to AMVP are myxomatous, bileaflet MVP and mitral annular disjunction (MAD). Data from autopsy, cardiac imaging and electrophysiological studies suggest that ectopy in AMVP is due to inflammation, fibrosis and scarring within the left ventricular (LV) base, LV papillary muscles and Purkinje tissue. Postulated mechanisms include repetitive injury to these regions from systolic papillary muscle stretch and abrupt mitral annular dysmotility (excursion and curling) and diastolic endocardial interaction of redundant mitral leaflets and chordae. Whereas AMVP is seen relatively commonly (up to 30%) in those with MVP, MVP-related SCD is rare (2–4%). However, the proportion at risk (i.e., with MMVP) is unknown. The clustering of cardiac morphological and electrophysiological characteristics similar to AMVP in otherwise idiopathic SCD suggests that MMVP arises when specific arrhythmia modulators allow for VF initiation and perpetuation through action potential prolongation, repolarization heterogeneity and Purkinje triggering. Adequately powered prospective studies are needed to assess strategies for identifying MMVP and the primary prevention of SCD, including ICD implantation, sympathetic modulation and early surgical mitral valve repair. Given the low event rate, a collaborative multicenter approach is essential.
Self-Injury in Adolescence Is Associated with Greater Behavioral Risk Avoidance, Not Risk-Taking
Authors: Dillahunt Alina K. , Feldman Daniel A. , Thomas Leah R. , Farstead Brian W. , Frandsen Summer B. , Lee Somi , Pazdera Myah , Galloway Jennica , Bessette Katie L. , Roberts Henrietta , Crowell Sheila E. , Watkins Edward R. , Langenecker Scott A. , Westlund Schreiner Melinda
Published: February 2022

Strategies to link impulsivity and self-injurious behaviors (SIBs) show highly variable results, and may differ depending on the impulsivity measure used. To better understand this lack of consistency, we investigated correlations between self-report and behavioral impulsivity, inhibitory control, SIBs, and rumination. We included participants aged 13–17 years with either current or remitted psychopathology who have (n = 31) and who do not have (n = 14) a history of SIBs. Participants completed self-report measures of impulsivity, the Rumination Responsiveness Scale (RRS), and two behavioral measures of impulsivity: the Balloon Analogue Risk Task (BART) and Parametric Go/No-Go (PGNG). Lifetime SIBs were positively associated with self-reported impulsivity, specifically positive and negative urgency. However, individuals with greater lifetime SIBs demonstrated greater risk aversion (lower impulsivity) as measured by the BART, whereas there was no relation between lifetime SIBs and PGNG performance. There was no relation between rumination and behavioral impulsivity, although greater rumination was associated with higher negative urgency. Future research examining the role of SIBs in the context of active versus remitted psychopathology is warranted. Because most adolescents were remitted from major depressive disorder at the time of study, follow-up studies can determine if lower risk-taking may aid individuals with more prior SIBs to achieve and maintain a remitted state.
Improved Kidney Allograft Function after Early Conversion of Fast IR-Tac Metabolizers to LCP-Tac
Authors: Thölking Gerold , Tosun-Koç Filiz , Jehn Ulrich , Koch Raphael , Pavenstädt Hermann , Suwelack Barbara , Reuter Stefan
Published: February 2022

Fast tacrolimus (Tac) metabolism is associated with a more rapid decline of renal function after renal transplantation (RTx). Because the pharmacokinetics of LCP-Tac (LCPT) and immediate-release Tac (IR-Tac) differ, we hypothesized that switching from IR-Tac to LCPT in kidney transplant recipients would improve the estimated glomerular filtration rate (eGFR), particularly in fast metabolizers. For proof of concept, we performed a pilot study including RTx patients who received de novo immunosuppression with IR-Tac. A Tac concentration-to-dose ratio (C/D ratio) < 1.05 ng/mL·1/mg defined fast metabolizers and ≥1.05 ng/mL·1/mg slow metabolizers one month after RTx. Patients were switched to LCPT ≥ 1 month after transplantation and followed for 3 years. Fast metabolizers (n = 58) were switched to LCPT earlier than slow metabolizers (n = 22) after RTx (2.0 (1.0–253.1) vs. 13.2 (1.2–172.8) months, p = 0.005). Twelve months after the conversion to LCPT, Tac doses were reduced by about 65% in both groups. The C/D ratios at 12 months had increased from 0.66 (0.24–2.10) to 1.74 (0.42–5.43) in fast and from 1.15 (0.32–3.60) to 2.75 (1.08–5.90) in slow metabolizers. Fast metabolizers showed noticeable recovery of mean eGFR already one month after the conversion (48.5 ± 17.6 vs. 41.5 ± 17.0 mL/min/1.73 m², p = 0.032) and at all subsequent time points, whereas the eGFR in slow metabolizers remained stable. Switching to LCPT increased Tac bioavailability, C/D ratio, and was associated with a noticeable recovery of renal function in fast metabolizers. Conversion to LCPT is safe and beneficial early after RTx.
Investigating the Long-Term Effect of an Interdisciplinary Multimodal Rehabilitation Program on Levels of Bioactive Lipids and Telomerase Activity in Blood from Patients with Chronic Pain
Authors: Stensson Niclas , Gerdle Björn , Rönne-Petersén Linn , Yang Liu L. , Lavebratt Catharina , Falkenberg Torkel , Ghafouri Bijar
Published: February 2022

Mechanism-based diagnosis and therapies for chronic pain are lacking. However, bio-psycho-social interventions such as interdisciplinary multimodal rehabilitation programs (IPRPs) have shown to be relatively effective treatments. In this context we aim to investigate the effects of IPRP on the changes in levels of bioactive lipids and telomerase activity in plasma, and if these changes are associated with changes in pain intensity and psychological distress. This exploratory study involves 18 patients with complex chronic pain participating in an IPRP. Self-reports of pain, psychological distress, physical activity, and blood samples were collected before the IPRP and at a six-month follow-up. Levels of arachidonoylethanolamide (AEA) and 2-arachidonoylglycerol (2-AG), palmitoylethanolamide (PEA), oleoylethanolamide (OEA), stearoylethanolamide (SEA), and telomerase activity were measured. Pain intensity was decreased, and SEA levels were increased at the six-month follow up. A significant correlation existed between changes in SEA levels and pain intensity. AEA levels, were inversely correlated with physical activity. Furthermore, 2-AG and telomerase activity was significantly correlated at the six-month follow-up. This study confirms that IPRP is relatively effective for reduction in chronic pain. Changes in SEA were correlated with changes in pain intensity, which might indicate that SEA changes reflect the pain reduction effects of IPRP.