The precursor transfer RNAs (pre-tRNAs) require extensive processing to generate mature tRNAs possessing proper fold, structural stability, and functionality required to sustain cellular viability. The road to tRNA maturation follows an ordered process: 5\'-processing, 3\'-processing, modifications at specific sites, if any, and 3\'-CCA addition before aminoacylation and recruitment to the cellular protein synthesis machinery. Ribonuclease P (RNase P) is a universally conserved endonuclease in all domains of life, performing the hydrolysis of pre-tRNA sequences at the 5\' end by the removal of phosphodiester linkages between nucleotides at position -1 and +1. Except for an archaeal species: Nanoarchaeum equitans where tRNAs are transcribed from leaderless-position +1, RNase P is indispensable for life and displays fundamental variations in terms of enzyme subunit composition, mechanism of substrate recognition and active site architecture, utilizing in all cases a two metal ion-mediated conserved catalytic reaction. While the canonical RNA-based ribonucleoprotein RNase P has been well-known to occur in bacteria, archaea, and eukaryotes, the occurrence of RNA-free protein-only RNase P in eukaryotes and RNA-free homologs of Aquifex RNase P in prokaryotes has been discovered more recently. This review aims to provide a comprehensive overview of structural diversity displayed by various RNA-based and RNA-free RNase P holoenzymes towards harnessing critical RNA-protein and protein-protein interactions in achieving conserved pre-tRNA processing functionality. Furthermore, alternate roles and functional interchangeability of RNase P are discussed in the context of its employability in several clinical and biotechnological applications. This article is categorized under: RNA Processing > tRNA Processing RNA Evolution and Genomics > RNA and Ribonucleoprotein Evolution RNA Interactions with Proteins and Other Molecules > RNA-Protein Complexes.

Protein-only RNase P (PRORP) is an essential enzyme responsible for the 5\' maturation of precursor tRNAs (pre-tRNAs). PRORPs are classified into three categories with unique molecular architectures, although all three classes of PRORPs share a mechanism and have similar active sites. Single subunit PRORPs, like those found in plants, have multiple isoforms with different localizations, substrate specificities, and temperature sensitivities. Most recently, Arabidopsis thaliana PRORP2 was shown to interact with TRM1A and B, highlighting a new potential role between these enzymes. Work with At PRORPs led to the development of a ribonuclease that is being used to protect against plant viruses. The mitochondrial RNase P complex, found in metazoans, consists of PRORP, TRMT10C, and SDR5C1, and has also been shown to have substrate specificity, although the cause is unknown. Mutations in mitochondrial tRNA and mitochondrial RNase P have been linked to human disease, highlighting the need to continue understanding this complex. The last class of PRORPs, homologs of Aquifex RNase P (HARPs), is found in thermophilic archaea and bacteria. This most recently discovered type of PRORP forms a large homo-oligomer complex. Although numerous structures of HARPs have been published, it is still unclear how HARPs bind pre-tRNAs and in what ratio. There is also little investigation into the substrate specificity and ideal conditions for HARPs. Moving forward, further work is required to fully characterize each of the three classes of PRORP, the pre-tRNA binding recognition mechanism, the rules of substrate specificity, and how these three distinct classes of PRORP evolved. This article is categorized under: RNA Structure and Dynamics > RNA Structure, Dynamics and Chemistry RNA Structure and Dynamics > Influence of RNA Structure in Biological Systems.

The endoribonuclease RNase P is responsible for tRNA 5\' maturation in all domains of life. A unique feature of RNase P is the variety of enzyme architectures, ranging from dual- to multi-subunit ribonucleoprotein forms with catalytic RNA subunits to protein-only enzymes, the latter occurring as single- or multi-subunit forms or homo-oligomeric assemblies. The protein-only enzymes evolved twice: a eukaryal protein-only RNase P termed PRORP and a bacterial/archaeal variant termed homolog of Aquifex RNase P (HARP); the latter replaced the RNA-based enzyme in a small group of thermophilic bacteria but otherwise coexists with the ribonucleoprotein enzyme in a few other bacteria as well as in those archaea that also encode a HARP. Here we summarize the history of the discovery of protein-only RNase P enzymes and review the state of knowledge on structure and function of bacterial HARPs and eukaryal PRORPs, including human mitochondrial RNase P as a paradigm of multi-subunit PRORPs. We also describe the phylogenetic distribution and evolution of PRORPs, as well as possible reasons for the spread of PRORPs in the eukaryal tree and for the recruitment of two additional protein subunits to metazoan mitochondrial PRORP. We outline potential applications of PRORPs in plant biotechnology and address diseases associated with mutations in human mitochondrial RNase P genes. Finally, we consider possible causes underlying the displacement of the ancient RNA enzyme by a protein-only enzyme in a small group of bacteria.

Natural gut harp strings are made from twisted bundles of gut strips, which are dried, ground, and varnished. The effects of varying the twist angle and surface finishing on the mechanical properties of gut harp strings have been explored. Strings were tested over a range of twist angles from 23.5∘ to 58.3∘, and with all four combinations of ground or unground and varnished or unvarnished surface finishing. The principal effects of varying the degree of twisting were that the breaking strength and tensile Young\'s modulus both fell as the twist angle was increased. String makers must therefore make a compromise between sound quality and string strength and durability. Leaving the string unvarnished dramatically increased the sensitivity to changes in humidity, which, in turn, affected the thermal tuning sensitivity and creep behaviour. Grinding the string surface prior to varnishing had no significant effect on the behaviour, but did make some difference to the thermal tuning sensitivity if the string was left unvarnished. Increasing the humidity frequently triggered episodes of additional string creep. There appeared to be a threshold effect, with the additional creep triggered when the string linear density exceeded its previous maximum. When the string was not creeping, there appeared to be a strong coupling mechanism between changes in the linear density and complementary changes in the string tension, such that there was almost no net effect on the string frequency. This behaviour was independent of the twist angle and the surface finishing, suggesting that whatever the coupling mechanism was, it was not dependent on the twisted structure of the string.

UNASSIGNED: Older adults admitted to a hospital for acute illness are at higher risk of hospital-associated functional decline during stays and after discharge.
UNASSIGNED: This study aimed to assess the calibration and discriminative abilities of the Hospital Admission Risk Profile (HARP) and the Identification of Seniors at Risk (ISAR) scales as predictors of hospital-associated functional decline at discharge in a cohort of patients older than age 65 receiving management in an acute geriatric care unit in Colombia.
UNASSIGNED: This study is an external validation of ISAR and HARP prediction models in a cohort of patients over 65 years managed in an acute geriatric care unit. The study included patients with Barthel index measured at admission and discharge. The evaluation discriminate ability and calibration, two fundamental aspects of the scales.
UNASSIGNED: Of 833 patients evaluated, 363 (43.6%) presented hospital-associated functional decline at discharge. The HARP underestimated the risk of hospital-associated functional decline for patients in low- and intermediate-risk categories (relation between observed/expected events (ROE) 1.82 and 1.51, respectively). The HARP overestimated the risk of hospital-associated functional decline for patients in the high-risk category (ROE 0.91). The ISAR underestimated the risk of hospital-associated functional decline for patients in low- and high-risk categories (ROE 1.59 and 1.11). Both scales showed poor discriminative ability, with an area under the curve (AUC) between 0.55 and 0.60.
UNASSIGNED: This study found that HARP and ISAR scales have limited discriminative ability to predict HAFD at discharge. The HARP and ISAR scales should be used cautiously in the Colombian population since they underestimate the risk of hospital-associated functional decline and have low discriminative ability.
UNASSIGNED: los adultos mayores ingresados en un hospital por una enfermedad aguda tienen un mayor riesgo de deterioro functional hospitalario durante su estancia y después del alta.
UNASSIGNED: este estudio tuvo como objetivo evaluar las capacidades de calibración y discriminación de las escalas Hospital Admission Risk Profile (HARP) e Identification of Seniors at Risk (ISAR) como predictores de deterioro funcional hospitalario al alta en una cohorte de pacientes mayores de 65 años que recibieron manejo en una unidad geriátrica de agudos en Colombia.
UNASSIGNED: este estudio es una validación externa de los modelos de predicción ISAR y HARP en una cohorte de pacientes mayores de 65 años atendidos en una unidad geriátrica de agudos. El estudio incluyó pacientes con índice de Barthel medido al ingreso y al alta y la evaluación de la capacidad de discriminación y calibración, dos aspectos fundamentales para esta medición.
UNASSIGNED: de 833 pacientes evaluados, 363 (43.6%) presentaron deterioro funcional hospitalario al momento del alta. La escala HARP subestimó el riesgo de deterioro funcional hospitalario para los pacientes en las categorías de riesgo bajo e intermedio (relación entre eventos observados /esperados (ROE) 1.82 y 1.51, respectivamente). El HARP sobrestimó el riesgo de deterioro funcional hospitalario para pacientes en la categoría de alto riesgo (ROE 0.91). El ISAR subestimó el riesgo de deterioro hospitalario para pacientes en categorías de bajo y alto riesgo (ROE 1.59 y 1.11). Ambas escalas mostraron una pobre capacidad de discriminación, con un área bajo la curva (AUC) entre 0.55 y 0.60.
UNASSIGNED: este estudio encontró que las escalas HARP e ISAR tienen una capacidad de discriminación limitada para predecir deterioro funcional hospitalario al alta. Las escalas HARP e ISAR deben usarse con cautela en la población colombiana ya que subestiman el riesgo de deterioro funcional hospitalario y tienen baja capacidad de discriminación.

A small group of bacteria encode two types of RNase P, the classical ribonucleoprotein (RNP) RNase P as well as the protein-only RNase P HARP (homolog of Aquifex RNase P). We characterized the dual RNase P activities of five bacteria that belong to three different phyla. All five bacterial species encode functional RNA (gene rnpB) and protein (gene rnpA) subunits of RNP RNase P, but only the HARP of the thermophile Thermodesulfatator indicus (phylum Thermodesulfobacteria) was found to have robust tRNA 5\'-end maturation activity in vitro and in vivo in an Escherichia coli RNase P depletion strain. These findings suggest that both types of RNase P are able to contribute to the essential tRNA 5\'-end maturation activity in T. indicus, thus resembling the predicted evolutionary transition state in the progenitor of the Aquificaceae before the loss of rnpA and rnpB genes in this family of bacteria. Remarkably, T. indicus RNase P RNA is transcribed with a P12 expansion segment that is posttranscriptionally excised in vivo, such that the major fraction of the RNA is fragmented and thereby truncated by ∼70 nt in the native T. indicus host as well as in the E. coli complementation strain. Replacing the native P12 element of T. indicus RNase P RNA with the short P12 helix of Thermotoga maritima RNase P RNA abolished fragmentation, but simultaneously impaired complementation efficiency in E. coli cells, suggesting that intracellular fragmentation and truncation of T. indicus RNase P RNA may be beneficial to RNA folding and/or enzymatic activity.

The rescue of stalled DNA replication forks is essential for cell viability. Impeded but still intact forks can be rescued by atypical DNA helicases in a reaction known as fork regression. This reaction has been studied at the single-molecule level using the Escherichia coli DNA helicase RecG and, separately, using the eukaryotic SMARCAL1 enzyme. Both nanomachines possess the necessary activities to regress forks: they simultaneously couple DNA unwinding to duplex rewinding and the displacement of bound proteins. Furthermore, they can regress a fork into a Holliday junction structure, the central intermediate of many fork regression models. However, there are key differences between these two enzymes. RecG is monomeric and unidirectional, catalyzing an efficient and processive fork regression reaction and, in the process, generating a significant amount of force that is used to displace the tightly-bound E. coli SSB protein. In contrast, the inefficient SMARCAL1 is not unidirectional, displays limited processivity, and likely uses fork rewinding to facilitate RPA displacement. Like many other eukaryotic enzymes, SMARCAL1 may require additional factors and/or post-translational modifications to enhance its catalytic activity, whereas RecG can drive fork regression on its own.

Three tree ring sequences were collected on the soundboard of the Stradivari harp. Due to the presence of the strings in the centre of the harp soundboard, the sampling of the tree ring widths was focused separately on the right side (RX), the central (CX) and the left side (LX). Tree ring measurements were carried out by using the Video Time Table (VTT), an instrument that combines a portable measuring device and a digital, high-resolution video camera. The VTT allowed non-invasive measurements of the tree rings to be made in situ and to immediately verify the quality of the sampling. The growth rings of the central portion were sampled using a high-resolution camera, which made it possible to bypass the barrier formed by the neck and strings. The consequent parallax and focus problems were overcome by taking many photographs from different angles. The measurements on the photographs were made with the CooRecorder program. The dendrochronological data were acquired with the PAST4 program and graphically processed and analysed with the PAST4 and 5 programs.

Ribonuclease P (RNase P) enzymes are responsible for the 5\' processing of tRNA precursors. In addition to the well-characterised ribozyme-based RNase P enzymes, an evolutionarily distinct group of protein-only RNase Ps exists. These proteinaceous RNase Ps (PRORPs) can be found in all three domains of life and can be divided into two structurally different types: eukaryotic and prokaryotic. Recent structural studies on members of both families reveal a surprising diversity of molecular architectures, but also highlight conceptual and mechanistic similarities. Here, we provide a comparison between the different types of PRORP enzymes and review how the combination of structural, biochemical, and biophysical studies has led to a molecular picture of protein-mediated tRNA processing.

We addressed comprehensively the performance of Shortest-Path HARP Refinement (SP-HR), SinMod, and DENSEanalysis using 2D slices of synthetic CSPAMM and DENSE images with realistic contrasts obtained from 3D phantoms. The three motion estimation techniques were interrogated under ideal and no-ideal conditions (with MR induced artifacts, noise, and through-plane motion), considering several resolutions and noise levels. Under noisy conditions, and for isotropic pixel sizes of 1.5 mm and 3.0 mm in CSPAMM and DENSE images respectively, the nRMSE obtained for the circumferential and radial strain components were 10.7 ± 10.8% and 25.5 ± 14.8% using SP-HR, 11.9 ± 2.5% and 29.3 ± 6.5% using SinMod, and 6.4 ± 2.0% and 18.2 ± 4.6% using DENSEanalysis. Overall, the results showed that SP-HR tends to fail for large tissue motions, whereas SinMod and DENSEanalysis gave accurate displacement and strain field estimations, being the last which performed the best.