Surface saturation of drug-loaded hollow manganese dioxide nanoparticles with human serum albumin for treating rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory joint disease accompanied by energy depletion and accumulation of reactive oxygen species (ROS). Inorganic nanoparticles (NPs) offer great promise for the treatment of RA because they mostly have functions beyond being drug carriers. However, conventional nanomaterials become coated with a protein corona (PC) or lose their cargo prematurely in vivo, reducing their therapeutic efficacy. To avoid these problems, we loaded methotrexate (MTX) into hollow structured manganese dioxide nanoparticles (H-MnO2 NPs), then coated them with a 'pseudo-corona' of human serum albumin (HSA) at physiological concentrations to obtain HSA-MnO2@MTX NPs. Efficacy of MTX, MnO2@MTX, and HSA-MnO2@MTX NPs was compared in vitro and in vivo. Compared to MnO2@MTX, HSA-coated NPs were taken up better by lipopolysaccharide-activated RAW264.7 and were more effective at lowering levels of pro-inflammatory cytokines and preventing ROS accumulation. HSA-MnO2@MTX NPs were also more efficient at blocking the proliferation and migration of fibroblast-like synoviocytes from rats with collagen-induced arthritis. In this rat model, HSA-MnO2@MTX NPs showed better biodistribution than other treatments, specifically targeting the ankle joint. Furthermore, HSA-MnO2@MTX NPs reduced swelling in the paw, regulated pro-inflammatory cytokine production, and limited cartilage degradation and signs of inflammation. These results establish the therapeutic potential of HSA-MnO2@MTX NPs against RA.

A Dual-Passband Frequency Selective Surface with High Angular Stability and Polarization Insensitivity.

In this paper, a dual-passband frequency selective surface (FSS) with high angular stability and polarization insensitivity is proposed. The unit structure consists of a circular aperture, two annular apertures and four cross apertures. The designed FSS can achieve a double-passband at the interested frequencies of 8.45 GHz and 12.76 GHz with an insertion loss of less than 1 dB, and it can retain a stable transmission characteristic with the incident angle ranging from 0° to 86° for TE mode and from 0° to 83° for TM mode. Good agreement between the experimental results and the simulated response verifies the feasibility of the proposed FSS.

The effect of macrophages and their exosomes in ischemic heart disease.

Ischemic heart disease (IHD) is a leading cause of disability and death worldwide, with immune regulation playing a crucial role in its pathogenesis. Various immune cells are involved, and as one of the key immune cells residing in the heart, macrophages play an indispensable role in the inflammatory and reparative processes during cardiac ischemia. Exosomes, extracellular vesicles containing lipids, nucleic acids, proteins, and other bioactive molecules, have emerged as important mediators in the regulatory functions of macrophages and hold promise as a novel therapeutic target for IHD. This review summarizes the regulatory mechanisms of different subsets of macrophages and their secreted exosomes during cardiac ischemia over the past five years. It also discusses the current status of clinical research utilizing macrophages and their exosomes, as well as strategies to enhance their therapeutic efficacy through biotechnology. The aim is to provide valuable insights for the treatment of IHD.

Application of Biotechniques for In Vitro Virus and Viroid Elimination in Pome Fruit Crops.

Sustainable production of pome fruit crops is dependent upon having virus-free planting materials. The production and distribution of plants derived from virus- and viroid-negative sources is necessary not only to control pome fruit viral diseases but also for sustainable breeding activities, as well as the safe movement of plant materials across borders. With variable success rates, different in vitro-based techniques, including shoot tip culture, micrografting, thermotherapy, chemotherapy, and shoot tip cryotherapy, have been employed to eliminate viruses from pome fruits. Higher pathogen eradication efficiencies have been achieved by combining two or more of these techniques. An accurate diagnosis that confirms complete viral elimination is crucial for developing effective management strategies. In recent years, considerable efforts have resulted in new reliable and efficient virus detection methods. This comprehensive review documents the development and recent advances in biotechnological methods that produce healthy pome fruit plants. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

A Quad-Band Highly Selective Frequency Selective Surface with Ultra-Wideband Rejection.

In this paper, a highly selective quad-band frequency selective surface (FSS) with ultra-wideband rejection is presented. The proposed FSS structure was developed by cascading five metallic layers by three thin dielectric substrates. The five metallic layers are composed of two bent slot layers, two metallic square rings, and a metal patch. The dimensions of the unit cell are 0.13λ0× 0.13λ0× 0.18λ0 (λ0 is the free-space wavelength at the first operating frequency). The proposed structure achieves four transmission bands and has two wide stop-bands located at 1 to 5.5 GHz and 14 to 40 GHz, with a suppressed transmission coefficient below -20 dB. In order to verify the simulation results, an FSS prototype was fabricated and measured. It can be observed that the measured results are in favorable agreement with the simulation results. Its multiple narrow passbands and highly selective and ultra-wideband rejection properties ensure that our design can play a significant role in narrowband antennas, spatial filters, and many other fields.

Advances in the study of exosomes derived from mesenchymal stem cells and cardiac cells for the treatment of myocardial infarction.

Acute myocardial infarction has long been the leading cause of death in coronary heart disease, which is characterized by irreversible cardiomyocyte death and restricted blood supply. Conventional reperfusion therapy can further aggravate myocardial injury. Stem cell therapy, especially with mesenchymal stem cells (MSCs), has emerged as a promising approach to promote cardiac repair and improve cardiac function. MSCs may induce these effects by secreting exosomes containing therapeutically active RNA, proteins and lipids. Notably, normal cardiac function depends on intracardiac paracrine signaling via exosomes, and exosomes secreted by cardiac cells can partially reflect changes in the heart during disease, so analyzing these vesicles may provide valuable insights into the pathology of myocardial infarction as well as guide the development of new treatments. The present review examines how exosomes produced by MSCs and cardiac cells may influence injury after myocardial infarction and serve as therapies against such injury. Video Abstract.

Recent progress in therapeutic strategies and biomimetic nanomedicines based on neutrophils for inflammation treatment.

Inflammation is a defensive response that helps repair and restore damaged tissue by activating immune and nonimmune cells against pathogens. However, inflammation may cause chronic disease if it persists for a long time. Neutrophils recruit immediately to the lesion site and regulate the inflammatory process when inflammation occurs. Therefore, neutrophil-mediated therapy has been considered as a promising strategy for inflammatory diseases and has been extensively studied. In this review, we summarize the recent research progress of neutrophil-based personalized treatment strategies for inflammation. We also review the research progress of various neutrophil-mediated drug-delivery systems in combination with the inflammatory microenvironment.

Introduction: Neutrophils are the most abundant white blood cells in humans. When inflammation occurs, neutrophils bind to ligands on endothelial cells through a variety of specific receptors on their surfaces and migrate from the bone marrow to the site of inflammation across endothelial cells, meaning that they are able to recruit to the lesion site at the first opportunity and regulate the inflammatory process. Neutrophil-based therapeutic strategies and drug-delivery systems have been extensively studied in inflammatory diseases, as specific protein receptors are retained on the neutrophil membrane, which is considered a promising approach for inflammatory therapy. Areas covered: This work focuses on the various mechanisms of neutrophil-based therapeutic strategies and reviews the current types and advantages of neutrophil-mediated drug-delivery systems for the treatment of inflammatory disorders. Summary: Neutrophil-mediated drug-delivery systems have a wider range of disease applications, and neutrophil-based therapeutic strategies can be used to treat inflammation through different pathways. Our review of the literature suggests that neutrophil-targeted nanoparticles are one of the most promising strategies for the treatment of inflammation, as they promote drug accumulation at the site of inflammation and limit adverse events, while prolonging drug circulation in the blood and improving drug stability.

Micrografting: An Old Dog Plays New Tricks in Obligate Plant Pathogens.

Micrografting, which was developed almost 50 years ago, has long been used for virus eradication, micropropagation, regeneration, rejuvenation, and graft compatibility. Recently, micrografting has been used for studies of long-distance trafficking and signaling of molecules between scions and rootstocks. The graft transmissiveness of obligate plant pathogens, such as viruses, viroids, and phytoplasmas, facilitated the use of micrografting to study biological indexing and pathogen transmission, pathogen-induced graft incompatibility, and screening for the pathogen resistance during the past 20 years. The present study provides comprehensive information on the latter subjects. Finally, prospects are proposed to direct further studies.

Long-Term Preservation of Plant Viruses in Cryopreserved Shoot Tips.

Availability of the methods for long-term virus preservation facilitates easy acquirement of viruses, which are needed in many basic and applied virological studies. Cryopreservation is currently considered an ideal means for long-term preservation of plant germplasm. Recent studies have shown that cryopreservation provided an efficient and reliable method for long-term preservation of plant viruses. Here, we describe the detailed procedures of droplet vitrification for long-term preservation of apple stem grooving virus (ASGV), which represents a type of viruses that can invade meristematic cells of the shoot tips, and potato leafroll virus (PLRV), which is a phloem-limited virus that does not infect the apical meristem. Shoot tip cryopreservation provides an advantageous strategy for the long-term preservation of plant viruses.

ROS-induced oxidative stress in plant cryopreservation: occurrence and alleviation.

MAIN CONCLUSION: Reactive oxygen species (ROS)-induced oxidative stress results in low success or even total failure of cryopreservation. Better understanding of how the plant establishes resistance/tolerance to ROS-induced oxidative stress facilitates developments of robust cryopreservation procedures. Cryopreservation provides a safe and efficient strategy for long-term preservation of plant genetic resources. ROS-induced oxidative stress caused damage to cells and reduced the ability of the plant to survive following cryopreservation, eventually resulting in low success or even total failure. This paper provides updated and comprehensive information obtained in the past decade, including the following: (1) ROS generations and adaptive responses of antioxidant systems during cryopreservation; (2) expressions of oxidative stress-associated genes and proteins during cryopreservation; (3) ROS-triggered programmed cell death (PCD) during cryopreservation; and (4) exogenous applications of enzymatic and non-enzymatic antioxidants in improving success of cryopreservation. Prospects for further studies are proposed. The goal of the present study was to facilitate better understanding of the mechanisms by which the plant establishes resistance/tolerance to oxidative stress during cryopreservation and promote further studies toward the developments of robust cryopreservation procedures and wider application of plant cryobiotechnology.

Epigenetic and Genetic Integrity, Metabolic Stability, and Field Performance of Cryopreserved Plants.

Cryopreservation is considered an ideal strategy for the long-term preservation of plant genetic resources. Significant progress was achieved over the past several decades, resulting in the successful cryopreservation of the genetic resources of diverse plant species. Cryopreservation procedures often employ in vitro culture techniques and require the precise control of several steps, such as the excision of explants, preculture, osmo- and cryoprotection, dehydration, freeze-thaw cycle, unloading, and post-culture for the recovery of plants. These processes create a stressful environment and cause reactive oxygen species (ROS)-induced oxidative stress, which is detrimental to the growth and regeneration of tissues and plants from cryopreserved tissues. ROS-induced oxidative stresses were documented to induce (epi)genetic and somatic variations. Therefore, the development of true-to-type regenerants of the source germplasm is of primary concern in the application of plant cryopreservation technology. The present article provides a comprehensive assessment of epigenetic and genetic integrity, metabolic stability, and field performance of cryopreserved plants developed in the past decade. Potential areas and the directions of future research in plant cryopreservation are also proposed.

BiVO4 Ceramic Photoanode with Enhanced Photoelectrochemical Stability.

Monoclinic bismuth vanadate (BiVO4) is an attractive material with which to fabricate photoanodes due to its suitable band structure and excellent photoelectrochemical (PEC) performance. However, the poor PEC stability originating from its severe photo-corrosion greatly restricts its practical applications. In this paper, pristine and Mo doped BiVO4 ceramics were prepared using the spark plasma sintering (SPS) method, and their photoelectrochemical properties as photoanodes were investigated. The as-prepared 1% Mo doped BiVO4 ceramic (Mo-BVO (C)) photoanode exhibited enhanced PEC stability compared to 1% Mo doped BiVO4 films on fluorine doped Tin Oxide (FTO) coated glass substrates (Mo-BVO). Mo-BVO (C) exhibited a photocurrent density of 0.54 mA/cm2 and remained stable for 10 h at 1.23 V vs. reversible hydrogen electrode (RHE), while the photocurrent density of the Mo-BVO decreased from 0.66 mA/cm2 to 0.11 mA/cm2 at 1.23 V vs. RHE in 4 h. The experimental results indicated that the enhanced PEC stability of the Mo-BVO (C) could be attributed to its higher crystallinity, which could effectively inhibit the dissociation of vanadium in BiVO4 during the PEC process. This work may illustrate a novel ceramic design for the improvement of the stability of BiVO4 photoanodes, and might provide a general strategy for the improvement of the PEC stability of metal oxide photoanodes.

Development and Validation of a Simoa Assay for Determination of Recombinant Batroxobin in Human Serum.

Recombinant batroxobin (S3101) is a thrombin-like serine protease that binds to fibrinogen or is taken up by the reticuloendothelial system. A literature survey showed no adequate method that could determine sufficient concentrations to evaluate pharmacokinetic parameters for phase I clinical studies. Therefore, a sensitive method is urgently needed to support the clinical pharmacokinetic evaluation of S3101. In this study, a sensitive bioanalytical method was developed and validated, using a Quanterix single molecular array (Simoa) assay. Moreover, to thoroughly assess the platform, enzyme-linked immunosorbent assay and electrochemiluminescence assay were also developed, and their performance was compared with that of this novel technology platform. The assay was validated in compliance with the current guidelines. Measurements with the Simoa assay were precise and accurate, presenting a valid assay range from 6.55 to 4000 pg/mL. The intra- and inter-run accuracy and precision were within -19.3% to 15.3% and 5.5% to 17.0%, respectively. S3101 was stable in human serum for 280 days at -20°C and -70°C, for 2 h prior to pre-treatment and 24 h post pre-treatment at room temperature (22°C-28°C), respectively, and after five and two freeze-thaw cycles at -70°C and -20°C, respectively. The Simoa assay also demonstrated sufficient dilution linearity, assay sensitivity, and parallelism for quantifying S3101 in human serum. The Simoa assay is a sensitive and adequate method for evaluating the pharmacokinetic parameters of S3101 in human serum.

Peptide-based 68Ga-PET radiotracer for imaging CD133 expression in colorectal cancer.

OBJECTIVE: CD133 is a demonstrated cancer stem cell marker. A small peptide LS7, screened by a phage display technique, was identified to specifically target CD133. The purpose of this study was to develop a novel and specific peptide-based PET imaging agent for CD133 imaging in colorectal cancer. METHODS: The peptide LS7 was conjugated with 1,4,7,20-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) and radiolabeled with 68Ga. The cellular uptake was assessed in vitro. In vivo small-animal PET/CT and ex vivo biodistribution evaluations were performed in mice bearing CD133-positive HCT116 and Lovo cell-derived tumors as well as CD133-negative DLD1 cell-derived tumors. Nonspecific uptake of the tracer in HCT116 cell-derived tumor cells and tumor models was determined by coincubation or coinjection with an excess of unlabeled DOTA-LS7 along with radiolabeled tracers. RESULTS: 68Ga-DOTA-LS7 was produced with 80.0% yield and the radiochemical purity was greater than 95.0%. In vitro, 68Ga-DOTA-LS7 was selectively taken up by HCT116 and Lovo cells but not by DLD1 cells. Small-animal PET/CT clearly revealed deposition of 68Ga-DOTA-LS7 in HCT116 and Lovo cell-derived tumors with excellent contrast. Biodistribution demonstrated that the tumor uptakes were 2.24 ± 0.16, 1.76 ± 0.42, and 0.69 ± 0.28% ID/g in HCT116, Lovo and DLD1 cell-derived tumors, respectively, at 90 min post-injection. Uptake of 68Ga-DOTA-LS7 in HCT116 tumors was significantly inhibited by coinjection of excess DOTA-LS7. CONCLUSION: Rapid tumor CD133 detection and selectivity were demonstrated in vitro and in vivo with PET using the specific CD133 binding peptide 68Ga-DOTA-LS7. A robust correlation was detected in vivo between tumor signals from mouse xenograft models with different cell lines and CD133 expression. The favorable characteristics of 68Ga-DOTA-LS7, such as convenient synthesis and specific uptake, warrant its further investigation for CD133 expression imaging.

Development and validation of a triplex assay to study an antibody cocktail against Ebola virus in cynomolgus serum.

Background: To support the rapid development of an antibody cocktail against Ebola virus and avoid unnecessary exposure to infectious environments, an automatic and fast turnover triplex assay was developed using Simoa® (Quanterix Corporation, MA, USA). Materials & methods: A robust triplex assay was developed and validated for simultaneous quantification of the antibody cocktail against Ebola virus in cynomolgus serum. Results: The assay had a quantitation range of 78.1-5000 ng/ml. The intra- and interassay precisions (%CV) were within 11.4 and 13.9%, and the accuracies (%RE) were within -10.8 to 6.8%, respectively. Cross-reactivity was evaluated, and the results met the acceptance criteria. Conclusion: The assay was successfully applied to a pharmacokinetics study following a single-dose intravenous administration of 10 mg/kg the antibody cocktail against Ebola virus to cynomolgus monkeys.

Concurrent ductal adenocarcinoma, pseudocyst, and neuroendocrine tumor of the pancreas: A case report.

INTRODUCTION: Pancreatic pseudocyst is one of the most common cystic lesions. It always occurs following pancreatitis and is rarely found in combination with pancreatic adenocarcinoma. The coexistence of exocrine and neuroendocrine tumors of the pancreas is also infrequent. We herein report a case of simultaneous occurrence of a pancreatic ductal adenocarcinoma (PDAC), pseudocyst, and neuroendocrine tumor (NET), showing a "side-by-side pattern." PATIENT CONCERN: A 74-year-old man was hospitalized for epigastric pain and poor appetite. He had no history of pancreatitis, alcohol consumption, or smoking. DIAGNOSIS AND INTERVENTION: Abdominal enhanced computed tomography and magnetic resonance imaging revealed a 15 × 8 cm cystic lesion with poor enhancement located in the tail of the pancreas. The distal aspect of the main pancreatic duct was dilated. The pancreatic parenchyma adjacent to the cystic lesion showed slightly heterogeneous enhancement on computed tomography and magnetic resonance imaging. Laboratory examination showed an elevated carbohydrate antigen 19-9 serum level. The patient was preoperatively diagnosed with intraductal papillary mucinous neoplasm and subsequently underwent laparotomy. During the operation, a hard white tumor measuring about 4 × 3 cm was palpated adjacent to the cystic lesion on the duodenal side, and a 0.6-cm nodule was simultaneously found in the pancreatic tail. Therefore, total pancreatectomy and splenectomy were performed. Histopathological examination showed that the tumor was PDAC with an adjacent pseudocyst, and the small nodule was suggestive of a NET. OUTCOMES: The patient survived without recurrence or metastasis in the follow-up visit 10 months after the operation and adjuvant chemotherapy. CONCLUSIONS: The concomitant occurrence of a PDAC, pseudocyst, and NET has not been previously reported. We suggest that if a pancreatic cyst is found, the coincidental occurrence of a malignant tumor should be considered, especially if the carbohydrate antigen 19-9 level is increased. Additionally, dilation of the pancreatic duct may be a diagnostic clue. Furthermore, the simultaneous occurrence of pancreatic endocrine and exocrine tumors is very uncommon. Preoperative diagnosis becomes difficult because of the lack of specific symptoms and radiological features.

Development, progress and future prospects in cryobiotechnology of Lilium spp.

Lilium is one of the most popular flower crops worldwide, and some species are also used as vegetables and medicines. The availability of and easy access to diverse Lilium genetic resources are essential for plant genetic improvements. Cryopreservation is currently considered as an ideal means for the long-term preservation of plant germplasm. Over the last two decades, great efforts have been exerted in studies of Lilium cryopreservation and progress has been made in the successful cryopreservation of pollen, seeds and shoot tips in Lilium. Genes that exist in Lilium, including those that regulate flower shape, color and size, and that are resistant to cold stress and diseases caused by fungi and viruses, provide a rich source of valuable genetic resources for breeding programs to create novel cultivars required by the global floriculture and ornamental markets. Successful cryopreservation of Lilium spp. is a way to preserve these valuable genes. The present study provides updated and comprehensive information about the development of techniques that have advanced Lilium cryopreservation. Further ideas are proposed to better direct future studies on Lilium cryobiotechnology.

Cryobiotechnology: A Double-Edged Sword for Obligate Plant Pathogens.

Pathogen-free stock plants are required as propagation materials in nurseries and healthy materials are needed in germplasm exchange between countries or regions through quarantine programs. In addition, plant gene banks also prefer to maintain pathogen-free germplasm collections. Shoot tip cryotherapy is a novel biotechnology method whereby cryopreservation methods are used to eradicate obligate pathogens from vegetatively propagated plants. Long-term preservation of pathogens is necessary in all types of virus-related basic research and applications such as antigen preparation for virus detection by immunology-based methods, production of plant-based vaccines, genetic transformation to produce virus-derived resistant transgenic plants, and bionanotechnology to produce nano drugs. Obligate plant pathogens such as viruses and viroids are intracellular parasites that colonize only living cells of the hosts. Therefore, their long-term preservation is difficult. Cryotreatments cannot completely eradicate the obligate pathogens that do not infect meristematic cells and certain proportions of plants recovered from cryotreatments are still pathogen-infected. Furthermore, cryotreatments often fail to eradicate the obligate pathogens that infect meristematic cells. Cryopreservation can be used for the long-term cryopreservation of the obligate plant pathogens. Thus, cryobiotechnology functions as a double-edged sword for plant pathogen eradication and cryopreservation. This review provides updated a synthesis of advances in cryopreservation techniques for eradication and cryopreservation of obligate plant pathogens.

In vitro tissue culture of apple and other Malus species: recent advances and applications.

MAIN CONCLUSION: Studies on the tissue culture of apple have allowed for molecular, biotechnological and applied breeding research to advance. In the past 8 years, over 100 papers advancing basic biology, genetic transformation and cryobiology have emerged. Apple (Malus × domestica Borkh.; Rosaceae) is an important fruit crop grown mainly in temperate regions of the world. In vitro tissue culture is a biotechnological technique that has been used to genetically improve cultivars (scions) and rootstocks. This updated review presents a synthesis of findings related to the tissue culture of apple and other Malus spp. between 2010 and 2018. Increasingly complex molecular studies that are examining the apple genome, for example, in a bid to identify the cause of epigenetic mutations and the role of transposable elements in this process would benefit from genetically stable source material, which can be produced in vitro. Several notable or curious in vitro culture methods have been reported to improve shoot regeneration and induce the production of tetraploids in apple cultivars and rootstocks. Existing studies have revealed the molecular mechanism underlying the inhibition of adventitious roots by cytokinin. The use of the plant growth correction factor allows hypothetical shoot production from leaf-derived thin cell layers relative to conventional leaf explants to be determined. This updated review will allow novices and established researchers to advance apple and Malus biotechnology and breeding programs.

Long-term preservation of potato leafroll virus, potato virus S, and potato spindle tuber viroid in cryopreserved shoot tips.

Availability of and easy access to diverse plant viruses and viroids is a prerequisite in applied and basic studies related to viruses and viroids. Long-term preservation of viruses and viroids is difficult. A protocol was described for long-term preservation of potato leafroll virus (PLRV), potato virus S (PVS), and potato spindle tuber viroid (PSTVd) in cryopreserved shoot tips of potato cv. Zihuabai. Shoot regrowth levels following cryopreservation were higher in 1.5 mm-shoot tips (58-60%) than in 0.5-mm-ones (30-38%). All shoots recovered from 0.5-mm-shoot tips were PVS- and PSTVd-preserved, but none of them were PLRV-preserved. Cryopreservation of 1.5-mm-shoot tips resulted in 35% and 100% of PLRV- and PVS- and PSTVd-preserved shoots. Studies on cell survival patterns and virus localization provided explanations to the varying PLRV-preservation frequencies produced by cryopreservation of the two sizes of shoot tips. Although micropropagation efficiencies were low after 12 weeks of subculture following cryopreservation, similar efficiencies were obtained after 16 weeks of subculture in pathogen-preserved shoots recovered from cryopreservation, compared with the diseased in vitro stock shoots (the control). Pathogen concentrations in the three pathogens-preserved shoots analyzed by qRT-PCR were similar to those in micropropagated shoots. The three pathogens cryopreserved in shoot tips were readily transmitted by grafting and mechanical inoculation to potato plants. PLRV, PVS, and PSTVd represent a diverse range of plant viruses and viroid in terms of taxonomy and infectious ability. Therefore, shoot tip cryopreservation opens a new avenue for long-term preservation of the virus and viroid.