Intrinsically disordered protein PID-2 modulates Z granules and is required for heritable piRNA-induced silencing in the Caenorhabditis elegans embryo.

In Caenorhabditis elegans, the piRNA (21U RNA) pathway is required to establish proper gene regulation and an immortal germline. To achieve this, PRG-1-bound 21U RNAs trigger silencing mechanisms mediated by RNA-dependent RNA polymerase (RdRP)-synthetized 22G RNAs. This silencing can become PRG-1-independent and heritable over many generations, a state termed RNA-induced epigenetic gene silencing (RNAe). How and when RNAe is established, and how it is maintained, is not known. We show that maternally provided 21U RNAs can be sufficient for triggering RNAe in embryos. Additionally, we identify PID-2, a protein containing intrinsically disordered regions (IDRs), as a factor required for establishing and maintaining RNAe. PID-2 interacts with two newly identified and partially redundant eTudor domain-containing proteins, PID-4 and PID-5. PID-5 has an additional domain related to the X-prolyl aminopeptidase APP-1, and binds APP-1, implicating potential N-terminal proteolysis in RNAe. All three proteins are required for germline immortality, localize to perinuclear foci, affect size and appearance of RNA inheritance-linked Z granules, and are required for balancing of 22G RNA populations. Overall, our study identifies three new proteins with crucial functions in C. elegans small RNA silencing.

Internet-Based Psychological Interventions during SARS-CoV-2 Pandemic: An Experience in South of Italy.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has led to an increasing demand for online psychological intervention. The aim of this study is to evaluate the efficacy of received support in internet-based psychological intervention group (I-IG) patients, compared with a wait-list control group (CG). The Impact of Event Scale­Revised, Patient Health Questionnaire 9-item and Generalized Anxiety Disorder scale 7-item were administered. After participants had used the internet-based solution, the System Usability Scale was administered. In total, 221 patients (194 patients supported by internet-based interventions and 27 patients supported onsite) were included in intervention group, and 194 patients were included in CG. In a 6-month follow-up, participants in the I-IG demonstrated significant improvements in terms of PTSD risk (p < 0.0001, d = 0.64), depression (p < 0.0001, d = 0.68), and anxiety (p < 0.0001, d = 1.33), compared to the CG. Significant improvements in onsite intervention group patients with a large to very large effect size of PTSD risk (p < 0.0001, d = 0.91), depression (p < 0.0001, d = 0.81), and anxiety (p < 0.0001, d = 1.62) were found. After internet-based solution use, I-IG patients reported a very high usability and functionality (72.87 ± 13.11) of online intervention. In conclusion, SARS-CoV-2-related mental health problems can be improved by internet-based psychological intervention. The usability and functionality evaluation of online solutions by technological tools showed very positive results for the I-IG patients.

Maternal piRNAs Are Essential for Germline Development following De Novo Establishment of Endo-siRNAs in Caenorhabditis elegans.

The Piwi-piRNA pathway represents a germline-specific transposon-defense system. C. elegans Piwi, prg-1, is a non-essential gene and triggers a secondary RNAi response that depends on mutator genes, endo-siRNAs (22G-RNAs), and the 22G-RNA-binding Argonaute protein HRDE-1. Interestingly, silencing of PRG-1 targets can become PRG-1 independent. This state, known as RNAe, is heritable and depends on mutator genes and HRDE-1. We studied how the transgenerational memory of RNAe and the piRNA pathway interact. We find that maternally provided PRG-1 is required for de novo establishment of 22G-RNA populations, especially those targeting transposons. Strikingly, attempts to re-establish 22G-RNAs in absence of both PRG-1 and RNAe memory result in severe germline proliferation defects. This is accompanied by a disturbed balance between gene-activating and -repressing 22G-RNA pathways. We propose a model in which CSR-1 prevents the loading of HRDE-1 and in which both PRG-1 and HRDE-1 help to keep mutator activity focused on the proper targets.

Reproducibility of a semi-automatic method for 6-point vertebral morphometry in a multi-centre trial.

PURPOSE: To evaluate the reproducibility of a semi-automated system for vertebral morphometry (MorphoXpress) in a large multi-centre trial. MATERIALS AND METHODS: The study involved 132 clinicians (no radiologist) with different levels of experience across 20 osteo-centres in Italy. All have received training in using MorphoXpress. An expert radiologist was also involved providing data used as standard of reference. The test image originate from normal clinical activity and represent a variety of normal, under and over exposed films, indicating both normal anatomy and vertebral deformities. The image was represented twice to the clinicians in a random order. Using the software, the clinicians initially marked the midpoints of the upper and lower vertebrae to include as many of the vertebrae (T5-L4) as practical within each given image. MorphoXpress performs the localisation of all morphometric points based on statistical model-based vision system. Intra-operator as well inter-operator measurement of agreement was calculated using the coefficient of variation and the mean and standard deviation of the difference of two measurements to check their agreement. RESULTS: The overall intra-operator mean differences in vertebral heights is 1.61+/-4.27% (1 S.D.). The overall intra-operator coefficient of variation is 3.95%. The overall inter-operator mean differences in vertebral heights is 2.93+/-5.38% (1 S.D.). The overall inter-operator coefficient of variation is 6.89%. CONCLUSIONS: The technology tested here can facilitate reproducible quantitative morphometry suitable for large studies of vertebral deformities.

Assessment of osteoporotic vertebral fractures using specialized workflow software for 6-point morphometry.

PURPOSE: To evaluate the time required, the accuracy and the precision of a model-based image analysis software tool for the diagnosis of osteoporotic fractures using a 6-point morphometry protocol. MATERIALS AND METHODS: Lateral dorsal and lumbar radiographs were performed on 92 elderly women (mean age 69.2+/-5.7 years). Institutional review board approval and patient informed consent were obtained for all subjects. The semi-automated and the manual correct annotations of 6-point placement were compared to calculate the time consumed and the accuracy of the software. Twenty test images were randomly selected and the data obtained by multiple perturbed initialisation points on the same image were compared to assess the precision of the system. RESULTS: The time requirement data of the semi-automated system (420+/-67 s) were statistically different (p<0.05) from that of manual placement (900+/-77 s). In the accuracy test, the mean reproducibility error for semi-automatic 6-point placement was 2.50+/-0.72% [95% CI] for the anterior-posterior reference and 2.16+/-0.5% [95% CI] for the superior-inferior reference. In the precision test the mean error resulted averaged over all vertebrae was 2.6+/-1.3% in terms of vertebral width. CONCLUSIONS: The technique is time effective, accurate and precise and can, therefore, be recommended in large epidemiological studies and pharmaceutical trials for reporting of osteoporotic vertebral fractures.