The influence of distal screw length on the primary stability of volar plate osteosynthesis-a biomechanical study

Background: Extensor tendon irritation is one of the most common complications following volar locking plate osteosynthesis (VLPO) for distal radius fractures. It is most likely caused by distal screws protruding the dorsal cortex. Shorter distal screws could avoid this, yet the influence of distal screw length on the primary stability in VLPO is unknown. The aim of this study was to compare 75 to 100 % distal screw lengths in VLPO. Methods: A biomechanical study was conducted on 11 paired fresh-frozen radii. HRpQCT scans were performed to assess bone mineral density (BMD) and bone mineral content (BMC). The specimens were randomized pair-wise into two groups: 100 % (group A) and 75 % (group B) unicortical distal screw lengths. A validated fracture model for extra-articular distal radius fractures (AO-23 A3) was used. Polyaxial volar locking plates were mounted, and distal screws was inserted using a drill guide block. For group A, the distal screw tips were intended to be flush or just short of the dorsal cortex. In group B, a target screw length of 75 % was calculated. The specimens were tested to failure using a displacement-controlled axial compression test. Primary biomechanical stability was assessed by stiffness, elastic limit, and maximum force as well as with residual tilt, which quantified plastic deformation. Results: Nine specimens were tested successfully. BMD and BMC did not differ between the two groups. The mean distal screw length of group A was 21.7 +/- 2.6 mm (range: 16 to 26 mm),for group B 16.9 +/- 1.9 mm (range: 12 to 20 mm). Distal screws in group B were on average 5.6 +/- 0.9 mm (range: 3 to 7 mm) shorter than measured. No significant differences were found for stiffness (706 +/- 103 N/mm vs. 660 +/- 124 N/mm),elastic limit (177 +/- 25 N vs. 167 +/- 36 N),maximum force (493 +/- 139 N vs. 471 +/- 149 N),or residual tilt (7.3 degrees +/- 0.7 degrees vs. 7.1 degrees +/- 1.3 degrees). Conclusion: The 75 % distal screw length in VLPO provides similar primary stability to 100 % unicortical screw length. This study, for the first time, provides the biomechanical basis to choose distal screws significantly shorter then measured

The influence of distal screw length on the primary stability of volar plate osteosynthesis-a biomechanical study

Background: Extensor tendon irritation is one of the most common complications following volar locking plate osteosynthesis (VLPO) for distal radius fractures. It is most likely caused by distal screws protruding the dorsal cortex. Shorter distal screws could avoid this, yet the influence of distal screw length on the primary stability in VLPO is unknown. The aim of this study was to compare 75 to 100 % distal screw lengths in VLPO. Methods: A biomechanical study was conducted on 11 paired fresh-frozen radii. HRpQCT scans were performed to assess bone mineral density (BMD) and bone mineral content (BMC). The specimens were randomized pair-wise into two groups: 100 % (group A) and 75 % (group B) unicortical distal screw lengths. A validated fracture model for extra-articular distal radius fractures (AO-23 A3) was used. Polyaxial volar locking plates were mounted, and distal screws was inserted using a drill guide block. For group A, the distal screw tips were intended to be flush or just short of the dorsal cortex. In group B, a target screw length of 75 % was calculated. The specimens were tested to failure using a displacement-controlled axial compression test. Primary biomechanical stability was assessed by stiffness, elastic limit, and maximum force as well as with residual tilt, which quantified plastic deformation. Results: Nine specimens were tested successfully. BMD and BMC did not differ between the two groups. The mean distal screw length of group A was 21.7 +/- 2.6 mm (range: 16 to 26 mm),for group B 16.9 +/- 1.9 mm (range: 12 to 20 mm). Distal screws in group B were on average 5.6 +/- 0.9 mm (range: 3 to 7 mm) shorter than measured. No significant differences were found for stiffness (706 +/- 103 N/mm vs. 660 +/- 124 N/mm),elastic limit (177 +/- 25 N vs. 167 +/- 36 N),maximum force (493 +/- 139 N vs. 471 +/- 149 N),or residual tilt (7.3 degrees +/- 0.7 degrees vs. 7.1 degrees +/- 1.3 degrees). Conclusion: The 75 % distal screw length in VLPO provides similar primary stability to 100 % unicortical screw length. This study, for the first time, provides the biomechanical basis to choose distal screws significantly shorter then measured

A Novel Mechanism of Programmed Cell Death in Bacteria by Toxin–Antitoxin Systems Corrupts Peptidoglycan Synthesis

Most genomes of bacteria contain toxin–antitoxin (TA) systems. These gene systems encode a toxic protein and its cognate antitoxin. Upon antitoxin degradation, the toxin induces cell stasis or death. TA systems have been linked with numerous functions, including growth modulation, genome maintenance, and stress response. Members of the epsilon/zeta TA family are found throughout the genomes of pathogenic bacteria and were shown not only to stabilize resistance plasmids but also to promote virulence. The broad distribution of epsilon/zeta systems implies that zeta toxins utilize a ubiquitous bacteriotoxic mechanism. However, whereas all other TA families known to date poison macromolecules involved in translation or replication, the target of zeta toxins remained inscrutable. We used in vivo techniques such as microscropy and permeability assays to show that pneumococcal zeta toxin PezT impairs cell wall synthesis and triggers autolysis in Escherichia coli. Subsequently, we demonstrated in vitro that zeta toxins in general phosphorylate the ubiquitous peptidoglycan precursor uridine diphosphate-N-acetylglucosamine (UNAG) and that this activity is counteracted by binding of antitoxin. After identification of the product we verified the kinase activity in vivo by analyzing metabolite extracts of cells poisoned by PezT using high pressure liquid chromatograpy (HPLC). We further show that phosphorylated UNAG inhibitis MurA, the enzyme catalyzing the initial step in bacterial peptidoglycan biosynthesis. Additionally, we provide what is to our knowledge the first crystal structure of a zeta toxin bound to its substrate. We show that zeta toxins are novel kinases that poison bacteria through global inhibition of peptidoglycan synthesis. This provides a fundamental understanding of how epsilon/zeta TA systems stabilize mobile genetic elements. Additionally, our results imply a mechanism that connects activity of zeta toxin PezT to virulence of pneumococcal infections. Finally, we discuss how phosphorylated UNAG likely poisons additional pathways of bacterial cell wall synthesis, making it an attractive lead compound for development of new antibiotics

Cooperative Interaction of Transcription Termination Factors with the RNA Polymerase II C-terminal Domain

Phosphorylation of the C-terminal domain of RNA polymerase II controls the co-transcriptional assembly of RNA processing and transcription factors. Recruitment relies on conserved CTDinteracting domains that recognize different CTD phosphoisoforms during the transcription cycle, but the molecular basis for their specificity remains unclear. We show that the CTD-interacting domains of two transcription termination factors, Rtt103 and Pcf11, achieve high affinity and specificity both by specifically recognizing the phosphorylated CTD and by cooperatively binding to neighboring CTD repeats. Single amino acid mutations at the protein-protein interface abolish cooperativity and affect recruitment at the 3′-end processing site in vivo. We suggest that this cooperativity provides a signal-response mechanism to ensure that its action is confined only to proper polyadenylation sites where Serine 2 phosphorylation density is highest

ε/ζ systems: their role in resistance, virulence, and their potential for antibiotic development

Cell death in bacteria can be triggered by activation of self-inflicted molecular mechanisms. Pathogenic bacteria often make use of suicide mechanisms in which the death of individual cells benefits survival of the population. Important elements for programmed cell death in bacteria are proteinaceous toxin–antitoxin systems. While the toxin generally resides dormant in the bacterial cytosol in complex with its antitoxin, conditions such as impaired de novo synthesis of the antitoxin or nutritional stress lead to antitoxin degradation and toxin activation. A widespread toxin–antitoxin family consists of the ε/ζ systems, which are distributed over plasmids and chromosomes of various pathogenic bacteria. In its inactive state, the bacteriotoxic ζ toxin protein is inhibited by its cognate antitoxin ε. Upon degradation of ε, the ζ toxin is released allowing this enzyme to poison bacterial cell wall synthesis, which eventually triggers autolysis. ε/ζ systems ensure stable plasmid inheritance by inducing death in plasmid-deprived offspring cells. In contrast, chromosomally encoded ε/ζ systems were reported to contribute to virulence of pathogenic bacteria, possibly by inducing autolysis in individual cells under stressful conditions. The capability of toxin–antitoxin systems to kill bacteria has made them potential targets for new therapeutic compounds. Toxin activation could be hijacked to induce suicide of bacteria. Likewise, the unique mechanism of ζ toxins could serve as template for new drugs. Contrarily, inhibition of virulence-associated ζ toxins might attenuate infections. Here we provide an overview of ε/ζ toxin–antitoxin family and its potential role in the development of new therapeutic approaches in microbial defense

Non-canonical 3′-5′ Extension of RNA with Prebiotically Plausible Ribonucleoside 2′,3′-Cyclic Phosphates

Ribonucleoside 2′,3′-cyclic phosphates (N>p’s) are generated by multiple prebiotically plausible processes and are credible building blocks for the assembly of early RNA oligomers. While N>p’s can be polymerized into short RNAs by non-enzymatic processes with variable efficiency and regioselectivity, no enzymatic route for RNA synthesis had been described. Here we report such a non-canonical 3′-5′ nucleotidyl transferase activity. We engineered a variant of the hairpin ribozyme to catalyze addition of all four N>p’s (2′,3′-cyclic A-, G-, U-, and CMP) to the 5′-hydroxyl termini of RNA strands with 5′ nucleotide addition enhanced in all cases by eutectic ice phase formation at −7 °C. We also observed 5′ addition of 2′,3′-cyclic phosphate-activated β-nicotinamide adenine dinucleotide (NAD>p) and ACA>p RNA trinucleotide, and multiple additions of GUCCA>p RNA pentamers. Our results establish a new mode of RNA 3′-5′ extension with implications for RNA oligomer synthesis from prebiotic nucleotide pools

Supporting data and code for: Ribozyme-mediated RNA synthesis and replication in a model Hadean microenvironment

Enzyme-catalysed replication of nucleic acid sequences is a prerequisite for the survival and evolution of biological entities. Before the advent of protein synthesis, genetic information was most likely stored in and replicated by RNA. However, experimental systems for sustained RNA-dependent RNA-replication are difficult to realise, in part due to the high thermodynamic stability of duplex products and the low chemical stability of catalytic RNAs. Using a derivative of a group I intron as a model for an RNA replicase, we show that heated air-water interfaces that are exposed to a plausible CO2-rich atmosphere enable sense and antisense RNA replication as well as template-dependent synthesis and catalysis of a functional ribozyme in a one-pot reaction. Both reactions are driven by autonomous oscillations in salt concentrations and pH, resulting from precipitation of acidified dew droplets, which transiently destabilise RNA duplexes. Our results suggest that an abundant Hadean microenvironment may have promoted both replication and synthesis of functional RNAs

Ribozyme activity modulates the physical properties of RNA–peptide coacervates

Condensed coacervate phases are now understood to be important features of modern cell biology, as well as valuable protocellular models in origin-of-life studies and synthetic biology. In each of these fields, the development of model systems with varied and tuneable material properties is of great importance for replicating properties of life. Here, we develop a ligase ribozyme system capable of concatenating short RNA fragments into long chains. Our results show that the formation of coacervate microdroplets with the ligase ribozyme and poly(L-lysine) enhances ribozyme rate and yield, which in turn increases the length of the anionic polymer component of the system and imparts specific physical properties to the droplets. Droplets containing active ribozyme sequences resist growth, do not wet or spread on unpassivated surfaces, and exhibit reduced transfer of RNA between droplets when compared to controls containing inactive sequences. These altered behaviours, which stem from RNA sequence and catalytic activity, constitute a specific phenotype and potential fitness advantage, opening the door to selection and evolution experiments based on a genotype–phenotype linkage