Comparison of the endocranial- and brain volumes in brachycephalic dogs, mesaticephalic dogs and Cavalier King Charles spaniels in relation to their body weight

BACKGROUND: A number of studies have attempted to quantify the relative volumes of the endocranial volume and brain parenchyma in association with the pathogenesis of the Chiari-like malformation (CLM) in the Cavalier King Charles spaniel (CKCS). In our study we examine the influence of allometric scaling of the brain and cranial cavity volume on morphological parameters in different dog breeds. MRI scans of 110 dogs (35 mesaticephalic dogs, 35 brachycephalic dogs, 20 CKCSs with SM, and 20 CKCSs without SM) have been used to create 3-dimensional volumetric models of skull and brain parts. Volumes were related to body weight calculating the adjusted means for different breeds. RESULTS: There was a strong global dependency of all volumes to body weight (P<0.0001). The adjusted means of the absolute and relative volumes of brain parenchyma and cranial compartments are not significantly larger in CKCSs in comparison to brachycephalic and mesaticephalic dogs. A difference in absolute or relative volumes between CKCSs with and without SM after relating these values to body weight could not be identified. The relative volume of the hindbrain parenchyma (caudal fossa parenchyma percentage) was larger in brachycephalic dogs than in CKCSs, without causing herniation or SM. CONCLUSION: An influence of body weight exist in dogs, which can be sufficiently large to render conclusions on the difference in volumes of the brain and skull unsafe unless some account of the body weight is taken in the analysis. The results of this study challenge the role of overcrowding for the development of SM in dogs

An intact ribose moiety at A2602 of 23S rRNA is key to trigger peptidyl-tRNA hydrolysis during translation termination

Peptide bond formation and peptidyl-tRNA hydrolysis are the two elementary chemical reactions of protein synthesis catalyzed by the ribosomal peptidyl transferase ribozyme. Due to the combined effort of structural and biochemical studies, details of the peptidyl transfer reaction have become increasingly clearer. However, significantly less is known about the molecular events that lead to peptidyl-tRNA hydrolysis at the termination phase of translation. Here we have applied a recently introduced experimental system, which allows the ribosomal peptidyl transferase center (PTC) to be chemically engineered by the introduction of non-natural nucleoside analogs. By this approach single functional group modifications are incorporated, thus allowing their functional contributions in the PTC to be unravelled with improved precision. We show that an intact ribose sugar at the 23S rRNA residue A2602 is crucial for efficient peptidyl-tRNA hydrolysis, while having no apparent functional relevance for transpeptidation. Despite the fact that all investigated active site residues are universally conserved, the removal of the complete nucleobase or the ribose 2′-hydroxyl at A2602, U2585, U2506, A2451 or C2063 has no or only marginal inhibitory effects on the overall rate of peptidyl-tRNA hydrolysis. These findings underscore the exceptional functional importance of the ribose moiety at A2602 for triggering peptide release

Revealing the elusive molecular biology of the vault RNA

Nicht-kodierende RNAs werden nicht in Proteine translatiert und führen ihre verschiedenen biologischen Funktionen durch z.B Regulation der Genexpression oder Modulation der Genomintegrität aus. Kürzlich konnten wir zeigen, dass die zur Klasse der nicht-kodierenden RNAs gehörenden vault-assozierten RNAs (vtRNA) signifikant hochreguliert werden, wenn humane B-Zellen mit dem Epstein Barr Virus (EBV) infiziert werden. Vault assoziierte RNAs sind in den sogenannten Vault Komplex integriert. Dieser 13 MDa große Komplex ist ein hohler und tonnenförmiger Ribo-nukleo-proteinkomplex. Das Major Vault Protein (MVP) ist der Hauptstrukturbestandteil des Vault Komplexes. Nur 5% der gesamten vtRNA sind direkt mit den Vault Partikel assoziiert. Es ist sehr wenig über die Funktion des Vault Partikles und der vtRNAs bekannt. Allerdings scheint die Hochregulierung der vtRNA nach EBV Infektion kausal verknüpft zu sein. Des weiteren konnten wir zeigen, dass die vtRNA1-1 Überexpression zu einer (dosisabhängigen) erhöhten Virusetablierung führt und vor programmiertem Zelltod schützt. Zusätzlich konnten wir die für die beobachtete Apoptoseresistenz verantwortliche vtRNA Domäne ermitteln. MVP Knock-down Experimente konnten diese anti-apoptose Effekte nicht beeinflussen. Somit scheint die vtRNA1-1 (und nicht der Vault Komplex) für die verbesserte virale Etablierung und Apoptoseresistenz in humanen B-Zellen zuständig zu sein.Non-coding RNAs (ncRNA) are not translated into proteins and exert their various biologic functions for example by regulating gene expression or modulating genome integrity. Recently, we demonstrated that the ncRNA class of vault-associated RNAs (vtRNA) were significantly up-regulated in human B cells upon Epstein-Barr virus (EBV) infection. Vault-associated RNAs serve as integral parts of the so-called vault complex, a large hollow barrel-shaped ribonucleo-protein complex with a size of 13 MDa. The major vault protein (MVP) is the major structural part of the complex whereas only 5% of the vtRNAs are directly associated to the vault particle. To date very little is known about the function(s) of the vault complex and its ncRNAs. The up-regulation of vault RNA seems to be causally linked to EBV infection.Here we could demonstrate that overexpression of the vtRNA1-1 led to an improved viral establishment and protected cells from undergoing general apoptosis in a dosage dependent manner. We further were able to identify the domain within the vault RNA sequence that is responsible for the observed phenotype. MVP knock-down experiments could not impair the apoptosis reistance effects. These data suggest that the vtRNA1-1 and not the MVP contribute to viral establishment and protection from apoptosis in human B cells.by Melanie Amort-AchmüllerAbweichender Titel laut Übersetzung der Verfasserin/des VerfassersZsfassung in dt. SpracheInnsbruck, Med. Univ., Diss., 2014OeBB(VLID)13309

Expression of the vault RNA protects cells from undergoing apoptosis

Non-protein-coding RNAs are a functionally versatile class of transcripts exerting their biological roles on the RNA level. Recently, we demonstrated that the vault complex-associated RNAs (vtRNAs) are significantly upregulated in Epstein-Barr virus (EBV)-infected human B cells. Very little is known about the function(s) of the vtRNAs or the vault complex. Here, we individually express latent EBV-encoded proteins in B cells and identify the latent membrane protein 1 (LMP1) as trigger for vtRNA upregulation. Ectopic expression of vtRNA1-1, but not of the other vtRNA paralogues, results in an improved viral establishment and reduced apoptosis, a function located in the central domain of vtRNA1-1. Knockdown of the major vault protein has no effect on these phenotypes revealing that vtRNA1-1 and not the vault complex contributes to general cell death resistance. This study describes a NF-κB-mediated role of the non-coding vtRNA1-1 in inhibiting both the extrinsic and intrinsic apoptotic pathways

An intact ribose moiety at A2602 of 23S rRNA is

key to trigger peptidyl-tRNA hydrolysis during translation terminatio

Skeletal muscle–derived cell implantation for the treatment of fecal incontinence : a randomized, placebo-controlled study

Background and Aims: Fecal incontinence (FI) improvement following injection of autologous skeletal muscle–derived cells has been previously suggested. This study aimed to test the efficacy and safety of said cells through a multicenter, placebo-controlled study, to determine an appropriate cell dose, and to delineate the target patient population that can most benefit from cell therapy. Methods: Patients experiencing FI for at least 6 months were randomized to receive a cell-free medium or low or high dose of cells. All patients received pelvic floor electrical stimulation before and after treatment. Incontinence episode frequency (IEF), FI quality of life, FI burden assessed on a visual analog scale, Wexner score, and parameters reflecting anorectal physiological function were all assessed for up to 12 months. Results: Cell therapy improved IEF, FI quality of life, and FI burden, reaching a preset level of statistical significance in IEF change compared with the control treatment. Post hoc exploratory analyses indicated that patients with limited FI duration and high IEF at baseline are most responsive to cells. Effects prevailed or increased in the high cell count group from 6 to 12 months but plateaued or diminished in the low cell count and control groups. Most physiological parameters remained unaltered. No unexpected adverse events were observed. Conclusions: Injection of a high dose of autologous skeletal muscle–derived cells followed by electrical stimulation significantly improved FI, particularly in patients with limited FI duration and high IEF at baseline, and could become a valuable tool for treatment of FI, subject to confirmatory phase 3 trial(s). (ClinicalTrialRegister.eu; EudraCT Number: 2010-021463-32)