Coupling of Tetrahymena ribosomal RNA splicing to beta-galactosidase expression in Escherichia coli

Splicing of the Tetrahymena ribosomal RNA precursor is mediated by the folded structure of the RNA molecule and therefore occurs in the absence of any protein in vitro. The Tetrahymena intervening sequence (IVS) has been inserted into the gene for the alpha-donor fragment of beta-galactosidase in a recombinant plasmid. Production of functional beta-galactosidase is dependent on RNA splicing in vivo in Escherichia coli. Thus RNA self-splicing can occur at a rate sufficient to support gene expression in a prokaryote, despite the likely presence of ribosomes on the nascent RNA. The beta-galactosidase messenger RNA splicing system provides a useful method for screening for splicing-defective mutations, several of which have been characterized.     

Progressive ataxia and seizures in a Cocker Spaniel: a new type of neurodegenerative disease with novel intra-neuronal inclusions

AIMS: To describe the histopathological lesions of a new canine disease characterised by progressive ataxia, head tremor and seizures, and to deduce the cause of the lesions.

METHODS: Formalin-fixed tissues were processed into paraffin wax and epoxy resin for light and transmission electron microscopy of variously stained tissue sections.

RESULTS: Significant lesions relevant to the disease were found only in the brain. They consisted of hypoplasia of the cerebellum and the presence of large pale inclusions in the perikaryon of neurons in the neocortex and in macrophages. The inclusion material was not compartmentalised and did not stain for carbohydrate, mucopolysaccharide or lipid. This material displaced nuclei to the periphery of the cells where they were seen as basophilic distorted crescent-shaped structures.

CONCLUSIONS: The inclusions were probably made of polymerised protein similar, though not identical, to those of Pick, Lewy and Collins bodies that characterise a variety of chronic neurodegenerative diseases of humans. A genetic basis to this disease was considered probable.     

Mycobacterium avium subsp. paratuberculosis Detected in the Reproductive Tract of Cows from an Infected Herd

Mycobacterium avium subspecies paratuberculosis (MAP), the causal agent of paratuberculosis, was detected by quantitative real‐time IS900 PCR in the follicular fluid from the reproductive tracts of cows originating from one infected herd. As well as being detected in follicular fluid of cows shedding bacteria in their faeces, MAP was also detected in the follicular fluid of one apparently healthy, non‐shedding individual cow. The finding of MAP in follicular fluid is unexpected and could contribute to the lower viability of embryos and resultant lower pregnancy rate. In addition to finding contaminated follicular fluid, vaginal and uterine flush fluids were determined to be positive for the presence of MAP in 75% and 56.3% of the time of the cattle currently shedding MAP in their faeces, respectively. The presence of MAP in different parts of the reproductive tract was seen in clinically as well as subclinically infected cows. These findings extend our currently scant and contradictory knowledge about the dissemination of MAP in the reproductive tract of female cattle.     

Eutypella canker of maple: first report from Germany and situation in Austria

Eutypella canker of maple, caused by Eutypella parasitica (which is native to North America), is reported for the first time from Germany. From 2013 to 2015, this perennial canker disease was recorded on 105 maple trees in Munich. Six maple species were affected: Acer pseudoplatanus, A. campestre, A. platanoides, A. cappadocicum, A. heldreichii ssp. trautvetteri and A. hyrcanum. Occurrence on the latter three species represents new host records for E. parasitica. In Austria, Eutypella canker was newly discovered on two trees at a second locality in 2011, and it is now known to occur on seven A. pseudoplatanus trees at two localities, which are separated nearly 150 km. A. pseudoplatanus was the most frequent host of E. parasitica in Munich and Austria, which is in agreement with previous studies in Europe. The identity of the causative pathogen as E. parasitica was verified by ITS rDNA sequencing of fungal cultures obtained from cankers in Munich and at both Austrian localities. The presence of large and old cankers in both countries suggests that introduction of E. parasitica dates back a long time, probably several decades. The new records of Eutypella canker in Germany and Austria show that the disease is more widely distributed in central Europe than previously recognized.     

Myxobolus erythrophthalmi sp. n. and Myxobolus shaharomae sp. n. (Myxozoa: Myxobolidae) from the internal organs of rudd, Scardinius erythrophthalmus (L.), and bleak, Alburnus alburnus (L.)

During a survey of myxosporean parasites of cyprinid fish in Hungary, infections caused by unknown Myxobolus spp. were found in the internal organs of rudd, Scardinius erythrophthalmus, and bleak, Alburnus alburnus . Small plasmodia developed in blood vessels of the kidney, liver, testes and intestinal wall. The parasites were studied on the basis of spore morphology and by histological and molecular methods. In most cases, plasmodia were surrounded by host tissue without a host reaction; however, in advanced cases, a connective tissue capsule was seen around plasmodia. Spores collected from the two fish species differed from each other and from the known Myxobolus spp. both in their morphology and 18S rDNA sequences. The two species, described as M. erythrophthalmi sp. n. from rudd and M. shaharomae sp. n. from bleak, are characterized by a specific histotropism to blood vessels, while the organ specificity involves the kidney and for the latter species, most internal organs.     

Infection of the Carpathian brook lamprey (Eudontomyzon danfordi Regan, 1911) with a dermocystid parasite in the Tisza River Basin,Hungary

The Carpathian brook lamprey (Eudontomyzon danfordi Regan, 1911) is an endemic protected species of Cephalaspidomorphi in the Carpathian Basin. No parasites have become known from these jawless vertebrates to date. Here, the authors describe an infection from a single specimen manifesting in protuberant skin cysts 7–10 mm in diameter, scattered on the body surface. Similar dermal infection was observed in 25 of the 274 lampreys recorded in the population survey. Skin cysts filled with round spore-like structures of a dermocystid parasite were found. These particles measured 8–14 µm in diameter and had an about 0.5 µm thick wall, and containing mainly a granular mass and a relatively scarce plasma. No hyphae were recorded. Despite conspicuous morphological changes in the skin, no inflammatory reactions were found. The molecular analysis of 18S rDNA showed similarity to dermocystid species of several fish species but differed from them approximately by 2%. This is the first record of a dermocystid parasite infecting a jawless vertebrate.     

Efficacy of intra-muscular analgesics for acute pain in sheep

The analgesic action of intramuscularly injected buprenorphine, methadone, flunixin meglumine and xylazine was examined in sheep, using algesimetry based on a leg withdrawal response to an electrical stimulus. No analgesic response was detected for buprenorphine, methadone or flunixin meglumine. Only the αaL2-adrenoceptor agonist, xylazine, produced an analgesic response. The current required to elicit a response increased by 170% (4.5±0.43 mA to 12.23±1.14 mA; mean±SE) after a dose of 0.05 mg/kg xylazine; by 180% (4.73±0.3 mA to 13.28±2.35 mA) after 0.1 mg/kg; and by 510% (4.52±0.29 mA to 27.63±3.89 mA) after 0.2 mg/kg. Intramuscular xylazine appears to be an effective analgesic agent for acute pain in the sheep and further investigation into ideal administration regimens and dosage may provide more detailed information on relationships between dose, analgesic and sedative effects. The findings also suggest that some common analgesic agents, and opioids in particular, may be ineffective for the management of acute pain in sheep and that any analgesic should be administered only on the basis of its proven efficacy in that species.     

Fluid dynamics in suspension-feeding blackfish

Measurements of flow patterns and water velocities inside the oral cavity of blackfish (Orthodon microlepidotus), made with a fiberoptic endoscope and thermistor flow probe, revealed that gill-arch structures act in blackfish as barriers that direct particle-laden water to the mucus-covered roof of the oral cavity, where particles are retained. Gill-arch structures have previously been assumed to be the site of particle retention in suspension-feeding fishes. Water does not pass between these structures in blackfish, and they do not serve as filters that separate particles from the water. These results emphasize the importance of directly assessing flow velocity and direction inside the oral cavity of vertebrate suspension feeders, particularly at the level of the filtering elements.     

A labile phosphodiester bond at the ligation junction in a circular intervening sequence RNA

The excised intervening sequence of the Tetrahymena ribosomal RNA precursor mediates its own covalent cyclization in the absence of any protein. The circular molecule undergoes slow reopening at a single phosphodiester bond, the one that was formed during cyclization. The resulting linear molecule has 5'-phosphate and 3'-hydroxyl termini; these are unusual products for RNA hydrolysis but are typical of the other reactions mediated by this molecule. The reopened circle retains cleavage-ligation activity, as evidenced by its ability to undergo another round of cyclization and reopening. The finding that an RNA molecule can be folded so that a specific phosphate can be strained or activated helps to explain how the activation energy is lowered for RNA self-splicing. The proposed mechanisms may be relevant to several other RNA cleavage reactions that are RNA-mediated.