The effect of cycloplegia in the accuracy of autorefraction, keratometry and axial length using the Myopia Master.

BACKGROUND: Assessing refractive errors under cycloplegia is recommended for paediatric patients; however, this may not always be feasible. In these situations, refraction has to rely on measurements made under active accommodation which may increase measurements variability and error. Therefore, evaluating the accuracy and precision of non-cycloplegic refraction and biometric measurements is clinically relevant. The Myopia Master, a novel instrument combining autorefraction and biometry, is designed for monitoring refractive error and ocular biometry in myopia management. This study assessed its repeatability and agreement for autorefraction and biometric measurements pre- and post-cycloplegia. METHODS: A prospective cross-sectional study evaluated a cohort of 96 paediatric patients that underwent ophthalmologic examination. An optometrist performed two repeated measurements of autorefraction and biometry pre- and post-cycloplegia. Test-retest repeatability (TRT) was assessed as differences between consecutive measurements and agreement as differences between post- and pre-cycloplegia measurements, for spherical equivalent (SE), refractive and keratometric J0/J45 astigmatic components, mean keratometry (Km) and axial length (AL). RESULTS: Cycloplegia significantly improved the SE repeatability (TRT, pre-cyclo: 0.65 D, post-cyclo: 0.31 D). SE measurements were more repeatable in myopes and emmetropes compared to hyperopes. Keratometry (Km) repeatability did not change with cycloplegia (TRT, pre-cyclo: 0.25 D, post-cyclo:0.27 D) and AL repeatability improved marginally (TRT, pre-cyclo: 0.14 mm, post-cyclo: 0.09 mm). Regarding pre- and post-cycloplegia agreement, SE became more positive by + 0.79 D, varying with refractive error. Myopic eyes showed a mean difference of + 0.31 D, while hyperopes differed by + 1.57 D. Mean keratometry, refractive and keratometric J0/J45 and AL showed no clinically significant differences. CONCLUSIONS: Refractive error measurements, using the Myopia Master were 2.5x less precise pre-cycloplegia than post-cycloplegia. Accuracy of pre-cycloplegic refractive error measurements was often larger than the clinically significant threshold (0.25 D) and was refractive error dependent. The higher precision compared to autorefraction measurements, pre- and post-cycloplegia agreement and refractive error independence of AL measurements emphasize the superiority of AL in refractive error monitoring.

Influence of anterior chamber depth and vault on anterior chamber angle morphology after phakic posterior chamber intraocular lens implantation.

PURPOSE: This study aims to investigate the influence of anterior chamber depth (ACD) and vault on the anterior chamber angle (ACA) morphology in myopic individuals implanted with posterior chamber phakic intraocular lenses. METHODS: This retrospective case series involved 231 eyes receiving a 13.2-mm implantable collamer lens (ICL). Preoperative anterior chamber anatomy was assessed using anterior segment optical coherence tomography (AS-OCT) and optical tomography, while postoperative evaluation employed AS-OCT. ACA morphology was characterized pre- and postoperatively through trabecular iris angle (TIA750), ACA distance opening (AOD750) and trabecular iris space area (TISA750). The influence of ACD and vault was examined by categorizing the sample into ACD (shallow, average and deep) and vault (low, optimal and high) groups. RESULTS: Preoperative ACA morphology varied based on ACD, with shallower ACDs exhibiting narrower TIA750, smaller AOD750 and TISA750. ICL implantation induced greater ACA narrowing more in the deep ACD group (TIA750 = 20.1 degrees; AOD750 = 0.82 mm and TISA750 = 0.44 mm2) compared to the shallow ACD group (TIA750 = 15.2 degrees; AOD750 = 0.44 mm and TISA750 = 0.21 mm2). Postoperatively, deeper ACDs showed larger ACAs. Increasing vault magnitude led to increased ACA narrowing, with the low vault group exhibiting smaller closure (TIA750 = 14.3 degrees; AOD750 = 0.56 mm and TISA750 = 0.29 mm2) compared to the high vault group (TIA750 = 20.8 degrees; AOD750 = 0.73 mm and TISA750 = 0.36 mm2). The magnitude of ACA narrowing associated with the vault had a consistent effect across different ACD groups. CONCLUSIONS: Posterior chamber intraocular lens implantation results in ACA narrowing, the extent of which is contingent upon preoperative anterior chamber and ACA morphology, with additional influence from vault magnitude.

Expression of symptoms elicited by a hammerhead viroid through RNA silencing is related to population bottlenecks in the infected host.

Chlorosis is frequently incited by viroids, small nonprotein-coding, circular RNAs replicating in nuclei (family Pospiviroidae) or chloroplasts (family Avsunviroidae). Here, we investigated how chrysanthemum chlorotic mottle viroid (CChMVd, Avsunviroidae) colonizes, evolves and initiates disease. Progeny variants of natural and mutated CChMVd sequence variants inoculated in chrysanthemum plants were characterized, and plant responses were assessed by molecular assays. We showed that: chlorotic mottle induced by CChMVd reflects the spatial distribution and evolutionary behaviour in the infected host of pathogenic (containing a UUUC tetranucleotide) and nonpathogenic (lacking such a pathogenic determinant) variants; and RNA silencing is involved in the initiation of the chlorosis in symptomatic leaf sectors through a viroid-derived small RNA containing the pathogenic determinant that directs AGO1-mediated cleavage of the mRNA encoding the chloroplastic transketolase. This study provides the first evidence that colonization of leaf tissues by CChMVd is characterized by segregating variant populations differing in pathogenicity and with the ability to colonize leaf sectors (bottlenecks) and exclude other variants (superinfection exclusion). Importantly, no specific pathogenic viroid variants were found in the chlorotic spots caused by chrysanthemum stunt viroid (Pospiviroidae), thus establishing a clear distinction on how members of the two viroid families trigger chlorosis in the same host.

Discussion on Secure Standard Network of Sensors Powered by Microbial Fuel Cells.

Everyday tasks use sensors to monitor and provide information about processes in different scenarios, such as monitoring devices in manufacturing or homes. Sensors need to communicate, with or without wires, while providing secure information. Power can be derived from various energy sources, such as batteries, electrical power grids, and energy harvesting. Energy harvesting is a promising way to provide a sustainable and renewable source to power sensors by scavenging and converting energy from ambient energy sources. However, low energy is harvested through these methods. Therefore, it is becoming a challenge to design and deploy wireless sensor networks while ensuring the sensors have enough power to perform their tasks and communicate with each other through careful management and optimization, matching energy supply with demand. For this reason, data cryptography and authentication are needed to protect sensor communication. This paper studies how energy harvested with microbial fuel cells can be employed in algorithms used in data protection during sensor communication.

Vault differences in eyes implanted with spherical and toric implantable collamer lenses: an inter-eye analysis.

PURPOSE: To determine the influence of implantable collamer lenses (ICL) geometry, i.e. spherical and toric on the vault, and report the refractive and visual outcomes of patients bilaterally implanted with the two ICL geometries. METHODS: This retrospective case series analysed 41 patients implanted with a spherical ICL (sICL) in one eye and an equal sized toric ICL (tICL) in the fellow eye. The anatomical and ICL-related parameters were assessed using anterior-segment optical coherence tomography (AS-OCT Visante, Zeiss Meditec AG) and optical tomography (Pentacam, OCULUS). The influence of the anatomical and ICL-related parameters on the vault was determined using generalised estimating equations (GEE) to incorporate inter-eye correlations. RESULTS: Postoperative spherical equivalent was within ± 0.50D in 66% and 83% of the eyes, respectively implanted with sICL and tICL. The efficacy index in the sICL group was 1.06 and 1.14 in the tICL group. The mean inter-eye vault difference was -1.46 µm, anatomical and ICL-related parameters showed similar associations with the vault for sICL and tICL. The GEE identified the ICL size minus the anterior chamber width, the ICL spherical power and ICL central thickness as significant factors influencing the vault. CONCLUSIONS: Spherical and toric ICL showed good efficacy for the correction of myopia and astigmatism. Patients implanted bilaterally with sICL and tICL tend to present similar vaults. The vault produced by both types of ICL was mainly regulated by the oversizing of the ICL. This suggests that the ICL geometry (spherical vs toric) is a factor with limited influence on the vault, thus the sizing method of a sICL and tICL should be similar.

Association between Clinical Vision Measures and Visual Perception and Soccer Referees' On-field Performance.

SIGNIFICANCE: The decisions taken by soccer officials are critically important to game management. Understanding the underlying processes that mediate expert performance in soccer refereeing may lead to a better standard of officiating. Vision is the dominant source of incoming information upon which officials rely to make their on-field decisions. PURPOSE: We tested the hypothesis that performance on generic tests of vision and visual perception predicts domain-specific performance in elite-level soccer referees (R) and assistant referees (AR). METHODS: We assessed the vision of R and AR who officiate at the highest level in Portugal. To be eligible for inclusion, R and AR had to have officiated for at least two consecutive seasons across the 2014/2015, 2015/2016, and 2016/2017 seasons. A single, rank-order list of the performance of eligible officials was created based on the rank-order list for each season that was made by the Portuguese Soccer Federation. Clinical vision measures included visual acuity and stereoacuity, and visual perception measures were gathered using the Test of Visual Perceptual Skills, Third Edition. RESULTS: A total of 59 officials participated (21 R, 38 AR), 17 of whom officiated at the international level. The R and AR groups did not differ in vision or visual perception measures. We found that better stereoacuity (P < .001) and visual memory (P = .001) are associated with a higher rank order of on-field performance after adjusting for the age, experience, the national/international status, and the regional affiliation of the officials. Together, these two measures explain 22% of the variance in rank-order performance. CONCLUSIONS: This is the first study to show a link between the vision of officials and their on-field performance. The origin and significance of these findings remain to be established, and further work is required to establish whether they are component skills in the domain of soccer refereeing.

Degradome Analysis of Tomato and Nicotiana benthamiana Plants Infected with Potato Spindle Tuber Viroid.

Viroids are infectious non-coding RNAs that infect plants. During infection, viroid RNAs are targeted by Dicer-like proteins, generating viroid-derived small RNAs (vd-sRNAs) that can guide the sequence specific cleavage of cognate host mRNAs via an RNA silencing mechanism. To assess the involvement of these pathways in pathogenesis associated with nuclear-replicating viroids, high-throughput sequencing of sRNAs and degradome analysis were carried out on tomato and Nicotiana benthamiana plants infected by potato spindle tuber viroid (PSTVd). Both hosts develop similar stunting and leaf curling symptoms when infected by PSTVd, thus allowing comparative analyses. About one hundred tomato mRNAs potentially targeted for degradation by vd-sRNAs were initially identified. However, data from biological replicates and comparisons between mock and infected samples reduced the number of bona fide targets-i.e., those identified with high confidence in two infected biological replicates but not in the mock controls-to only eight mRNAs that encode proteins involved in development, transcription or defense. Somewhat surprisingly, results of RT-qPCR assays revealed that the accumulation of only four of these mRNAs was inhibited in the PSTVd-infected tomato. When these analyses were extended to mock inoculated and PSTVd-infected N. benthamiana plants, a completely different set of potential mRNA targets was identified. The failure to identify homologous mRNA(s) targeted by PSTVd-sRNA suggests that different pathways could be involved in the elicitation of similar symptoms in these two species. Moreover, no significant modifications in the accumulation of miRNAs and in the cleavage of their targeted mRNAs were detected in the infected tomato plants with respect to the mock controls. Taken together, these data suggest that stunting and leaf curling symptoms induced by PSTVd are elicited by a complex plant response involving multiple mechanisms, with RNA silencing being only one of the possible components.

Revisiting the cysteine-rich proteins encoded in the 3'-proximal open reading frame of the positive-sense single-stranded RNA of some monopartite filamentous plant viruses: functional dissection of p15 from grapevine virus B.

A reexamination of proteins with conserved cysteines and basic amino acids encoded by the 3'-proximal gene of the positive-sense single-stranded RNA of some monopartite filamentous plant viruses has been carried out. The cysteines are involved in a putative Zn-finger domain, which, together with the basic amino acids, form part of the nuclear or nucleolar localization signals. An in-depth study of one of these proteins, p15 from grapevine B virus (GVB), has shown: (i) a three-dimensional structure with four α-helices predicted by two independent in silico approaches, (ii) the nucleolus as the main accumulation site by applying confocal laser microscopy to a fusion between p15 and the green fluorescent protein, (iii) the involvement of the basic amino acids and the putative Zn-finger domain, mapping at the N-terminal region of p15, in the nucleolar localization signal, as revealed by the effect of six alanine substitution mutations, (iv) the p15 suppressor function of sense-mediated RNA silencing as revealed by agroinfiltration in a transgenic line of Nicotiana benthamiana, and (v) the enhancer activity of p15 on viral pathogenicity in N. benthamiana when expressed from a potato virus X vector. In addition, we elaborate on an evolutionary scenario for these filamentous viruses, invoking takeover by a common ancestor(s) of viral or host genes coding for those cysteine-rich proteins, followed by divergence, which would also explain why they are encoded in the 3'-proximal gene of the genomic single-stranded viral RNA.

Superparamagnetic Nanoparticles with Efficient Near-Infrared Photothermal Effect at the Second Biological Window.

Superparamagnetic nanoparticles (iron oxide nanoparticles-IONs) are suitable for hyperthermia after irradiating with radiofrequency radiation. Concerning the suitability for laser ablation, IONs present a low molar absorption coefficient in the near-infrared region close to 800 nm. For this reason, they are combined with other photothermal agents into a hybrid composite. Here, we show that IONs absorb and convert into heat the infrared radiation characteristic of the so-called second-biological window (1000-1350 nm) and, in consequence, they can be used for thermal ablation in such wavelengths. To the known excellent water solubility, colloidal stability and biocompatibility exhibited by IONs, an outstanding photothermal performance must be added. For instance, a temperature increase of 36 °C was obtained after irradiating at 8.7 W cm-2 for 10 min a suspension of IONs at iron concentration of 255 mg L-1. The photothermal conversion efficiency was ~72%. Furthermore, IONs showed high thermogenic stability during the whole process of heating/cooling. To sum up, while the use of IONs in the first bio-window (700-950 nm) presents some concerns, they appear to be good photothermal agents in the second biological window.

Viroids: survivors from the RNA world?

Because RNA can be a carrier of genetic information and a biocatalyst, there is a consensus that it emerged before DNA and proteins, which eventually assumed these roles and relegated RNA to intermediate functions. If such a scenario--the so-called RNA world--existed, we might hope to find its relics in our present world. The properties of viroids that make them candidates for being survivors of the RNA world include those expected for primitive RNA replicons: (a) small size imposed by error-prone replication, (b) high G + C content to increase replication fidelity, (c) circular structure for assuring complete replication without genomic tags, (d) structural periodicity for modular assembly into enlarged genomes, (e) lack of protein-coding ability consistent with a ribosome-free habitat, and (f) replication mediated in some by ribozymes, the fingerprint of the RNA world. With the advent of DNA and proteins, those protoviroids lost some abilities and became the plant parasites we now know.

How sequence variants of a plastid-replicating viroid with one single nucleotide change initiate disease in its natural host.

Understanding how viruses and subviral agents initiate disease is central to plant pathology. Whether RNA silencing mediates the primary lesion triggered by viroids (small non-protein-coding RNAs), or just intermediate-late steps of a signaling cascade, remains unsolved. While most variants of the plastid-replicating peach latent mosaic viroid (PLMVd) are asymptomatic, some incite peach mosaics or albinism (peach calico, PC). We have previously shown that two 21-nt small RNAs (PLMVd-sRNAs) containing a 12-13-nt PC-associated insertion guide cleavage, via RNA silencing, of the mRNA encoding a heat-shock protein involved in chloroplast biogenesis. To gain evidence supporting that such event is the initial lesion, and more specifically, that different chloroses have different primary causes, here we focused on a PLMVd-induced peach yellow mosaic (PYM) expressed in leaf sectors interspersed with others green. First, sequencing PLMVd-cDNAs from both sectors and bioassays mapped the PYM determinant at one nucleotide, a notion further sustained by the phenotype incited by other natural and artificial PLMVd variants. And second, sRNA deep-sequencing and RNA ligase-mediated RACE identified one PLMVd-sRNA with the PYM-associated change that guides cleavage, as predicted by RNA silencing, of the mRNA encoding a thylakoid translocase subunit required for chloroplast development. RT-qPCR showed lower accumulation of this mRNA in PYM-expressing tissues. Remarkably, PLMVd-sRNAs triggering PYM and PC have 5'-terminal Us, involving Argonaute 1 in what likely are the initial alterations eliciting distinct chloroses.

Phloem restriction of viroids in three citrus hosts is overcome by grafting with Etrog citron: potential involvement of a translocatable factor.

Viroid systemic spread involves cell-to-cell movement from initially infected cells via plasmodesmata, long-distance movement within the phloem and again cell-to-cell movement to invade distal tissues including the mesophyll. Citrus exocortis viroid (CEVd), hop stunt viroid, citrus bent leaf viroid, citrus dwarfing viroid, citrus bark cracking viroid and citrus viroid V remained phloem restricted when singly infecting Citrus karna, Citrus aurantium and Poncirus trifoliata, but not Etrog citron, where they were additionally detected in mesophyll protoplasts. However, when CEVd-infected C. karna was side-grafted with Etrog citron--with the resulting plants being composed of a C. karna stock and an Etrog citron branch--the viroid was detected in mesophyll protoplasts of the former, thus indicating that the ability of Etrog citron to support viroid invasion of non-vascular tissues was transferred to the stock. Further results suggest that a translocatable factor from Etrog citron mediates this viroid trafficking.

Virus-viroid interactions: Citrus Tristeza Virus enhances the accumulation of Citrus Dwarfing Viroid in Mexican lime via virus-encoded silencing suppressors.

An assay to identify interactions between Citrus Dwarfing Viroid (CDVd) and Citrus Tristeza Virus (CTV) showed that viroid titer was enhanced by the coinfecting CTV in Mexican lime but not in etrog citron. Since CTV encodes three RNA silencing suppressors (RSSs), p23, p20 and p25, an assay using transgenic Mexican limes expressing each RSS revealed that p23 and, to a lesser extent, p25 recapitulated the effect observed with coinfections of CTV and CDVd.

Dahlia latent viroid: a recombinant new species of the family Pospiviroidae posing intriguing questions about its origin and classification.

A viroid-like RNA has been detected in two asymptomatic dahlia accessions by return and double PAGE. It appeared smaller than Chrysanthemum stunt viroid and Potato spindle tuber viroid, the two members of the genus Pospiviroid, family Pospiviroidae, reported in this ornamental previously. RT-PCR with primers designed for amplifying all pospiviroids produced no amplicons, but RT-PCR with random primers revealed a 342 nt RNA. The sequence of this RNA was confirmed with specific primers, which additionally revealed its presence in many dahlia cultivars. The RNA was named Dahlia latent viroid (DLVd) because it replicates autonomously, but symptomlessly, in dahlia and shares maximum sequence identity with other viroids of less than 56 %. Furthermore, DLVd displays characteristic features of the family Pospiviroidae: a predicted rod-like secondary structure of minimum free energy with a central conserved region (CCR), and the ability to form the metastable structures hairpins I and II. Its CCR is identical to that of Hop stunt viroid (HSVd, genus Hostuviroid). However, DLVd: (i) has the terminal conserved region present in members of the genus Pospiviroid, but absent in HSVd, and (ii) lacks the terminal conserved hairpin present in HSVd. Phylogenetic reconstructions indicate that HSVd and Pepper chat fruit viroid (genus Pospiviroid) are the closest relatives of DLVd, but DLVd differs from these viroids in its host range, restricted to dahlia so far. Therefore, while DLVd fulfils the criteria to be a novel species of the family Pospiviroidae, its recombinant origin makes assignment to the genera Pospiviroid or Hostuviroid problematic.

Repeatability and agreement of swept-source optical coherence tomography and partial coherence interferometry biometers in myopes.

CLINICAL RELEVANCE: Biometric measurements in the context of myopia are fundamental to detect eyes at risk of developing myopia and during the follow-up of patients with myopia control treatment. Thus, the accuracy of biometers has high clinical relevance. BACKGROUND: The Myopia Master is a new biometer based on partial coherence interferometry especially dedicated to the follow-up of myopic patients. This study aims to assess the repeatability of the Myopia Master and evaluate its agreement with a swept-source optical coherence interferometry biometer (IOL Master 700). METHODS: This cross-sectional prospective study assessed the biometric parameters of two groups of myopes (age range: 8-16 years old), spectacle corrected (n = 60) and orthokeratology contact lens wearers (n = 60). One senior optometrist performed two consecutive measurements per instrument, which included axial length, mean keratometry and horizontal visible iris diameter (HVID). The repeatability of each device and the agreement between devices were assessed by the dispersion of the measurement differences, for AL, mean keratometry, corneal astigmatism and HVID. RESULTS: The two biometers measured approximately the same value in both measurements. Test-retest repeatability tended to be lower than clinical significant thresholds, in particular, for AL and mean keratometry. Corneal-related parameters tended to have lower repeatability in the orthokeratology group, especially mean keratometry. The agreement between instruments revealed statistically significant differences between devices with the SS-OCT measuring longer eyes, steeper corneas and larger HVID. CONCLUSIONS: In a paediatric population, the Myopia Master showed clinically acceptable repeatability levels, but the IOL Master 700 demonstrated superior repeatability. Eyes treated with orthokeratology may compromise the repeatability of the corneal-related parameters. The Myopia Master and the IOL Master 700 are repeatable devices appropriate for monitoring myopia progression, but the differences observed do not allow their use interchangeably.

Prediction model of the horizontal trabecular iris angle after phakic posterior chamber implantable intraocular lens surgery.

PURPOSE: To determine the predictors of the postoperative horizontal trabecular iris angle (TIA 750 ) after phakic posterior chamber implantable intraocular lens (IOL) surgery. SETTING: Ophthalmology Clinic Vista Sánchez Trancón, Badajoz, Spain. DESIGN: Retrospective case series. METHODS: 330 eyes implanted with spherical/toric implantable collamer lens (ICL) were included in this study. From 230 eyes implanted with 13.2 mm ICL, these were divided in modeling (n = 180) and evaluation group (n = 50). Two groups implanted with 12.6 mm and 13.7 mm (n = 50 each) were also used as evaluation. Anterior-segment optical coherence tomography was used preoperatively to perform anterior chamber biometry (angle-to-angle [ATA] distance, crystalline lens rise, anterior chamber depth [ACD], cornea sagittal depth, pupil diameter, nasal/temporal TIA 750 ); postoperatively for measuring the vault, pupil diameter and nasal/temporal TIA 750 . Corneal curvature and horizontal visible iris diameter were measured using optical tomography. Bivariate correlation analysis was used to determine associations between preoperative and postoperative horizontal TIA 750 with anterior chamber biometry, ICL-related parameters and age. Finally, a multivariate linear regression model was constructed for predicting the postoperative TIA 750 . RESULTS: Horizontal TIA 750 reduced from 42.9 ± 8.0 degrees preoperatively to 24.4 ± 5.6 degrees postoperatively. Postoperative TIA 750 was positively correlated with the preoperative TIA 750 , cornea sagittal depth and ACD, and negatively associated with the vault. The main predictors of the postoperative TIA 750 were the preoperative parameters, TIA 750 , ICLsize - ATA and pupil diameter (adjusted- R2 = 0.39). The limits of agreement between predicted and real TIA 750 were close to ±10 degrees. CONCLUSIONS: Implantation of a phakic posterior chamber implantable IOL leads to a reduction in TIA 750 and the main factors contributing for this are the preoperative TIA 750 aperture and the vault.

Role of RNA silencing in plant-viroid interactions and in viroid pathogenesis.

Viroids are small, single-stranded, non-protein coding and circular RNAs able to infect host plants in the absence of any helper virus. They may elicit symptoms in their hosts, but the underlying molecular pathways are only partially known. Here we address the role of post-transcriptional RNA silencing in plant-viroid-interplay, with major emphasis on the involvement of this sequence-specific RNA degradation mechanism in both plant antiviroid defence and viroid pathogenesis. This review is a tribute to the memory of Dr. Ricardo Flores, who largely contributed to elucidate this and other molecular mechanisms involved in plant-viroid interactions.

Viroids and hepatitis delta virus.

There is a subviral world, whose most prominent representatives are viroids. Despite being solely composed by a circular, highly structured RNA of ~250 to 400 nucleotides without protein-coding ability (all viruses code for one or more proteins), viroids can infect and incite specific diseases in higher plants. The RNA of human hepatitis delta virus (HDV), the smallest genome of an animal virus, displays striking similarities with viroids: It is circular, folds into a rodlike secondary structure, and replicates through a rolling-circle mechanism catalyzed by host enzymes and cis-acting ribozymes. However, HDV RNA is larger (~1700 nucleotides), encodes a protein in its antigenomic polarity (the ∂ antigen), and depends for transmission on hepatitis B virus. The presence of ribozymes in some viroids and in HDV RNA, along with their structural simplicity, makes them candidates for being molecular fossils of the RNA world that presumably preceded our extant world based on DNA and proteins.

Gel Blot Hybridization for Viroids.

Northern blot analysis reveals information about RNA identity, size, and abundance. This technique has become an essential tool for the knowledge developed about viroids and also an excellent method for viroid detection. Here we describe the methodology of a Northern blot based in polyacrylamide gel electrophoresis under denaturing conditions, hybridized with a viroid full-length riboprobe labeled with chemiluminescence. Viroid detection with this approach entails positive signals, specific migration, and the differentiation of their circular and linear forms.