Reversal of dyskinesis by increased end diastolic segment length in ischaemic-reperfused myocardium.

Persistent dyskinesis is universally observed after reperfusion of a severely ischaemic segment. Although inotropic stimulation shows a latent contractile reserve, it is not known whether this reserve can be recruited by increasing end diastolic segment length (local length-tension relation). To investigate this, six anaesthetised open chest dogs were placed on right heart bypass to increase end diastolic segment length independently of mean arterial pressure. Instantaneous left ventricular pressure-segment length relations and fractional systolic shortening were determined by sonomicrometry in the centre of the region perfused by the left anterior descending coronary artery during sequential increases in end diastolic segment length. Measurements were made before occlusion of the left anterior descending coronary artery, during 1 h of occlusion, and after 2 h of reperfusion. Before ischaemia, segmental shortening increased from 11.0(SEM 1.6)% to 23.5(1.5)% (p less than 0.05) as end diastolic segment length increased. Dyskinesis developed during occlusion of the left anterior descending coronary artery [12.1(2.6)% control v -7.2(1.6)% occlusion, p less than 0.05] and was present over the entire range of end diastolic segment lengths. Following reperfusion, segmental dyskinesis [-2.5(2.4)%] persisted at the lower end of the range of end diastolic segment length, but was progressively replaced by active shortening, averaging 7.3(3.2)% (p less than 0.05) as end diastolic segment length was sequentially increased. We conclude that segmental function following reperfusion is sensitive to changes in end diastolic segment length, and that active shortening is recruited from an apparently dyskinetic segment as end diastolic segment length progressively increases.

Ruthenium-Caged Antisense Morpholinos for Regulating Gene Expression in Zebrafish Embryos.

Photochemical approaches afford high spatiotemporal control over molecular structure and function, for broad applications in materials and biological science. Here, we present the first example of a visible light responsive ruthenium-based photolinker, Ru(bipyridine)2(3-ethynylpyridine)2 (RuBEP), which was reacted stoichiometrically with a 25mer DNA or morpholino (MO) oligonucleotide functionalized with 3' and 5' terminal azides, via Cu(I)-mediated [3+2] Huisgen cycloaddition reactions. RuBEP-caged circular morpholinos (Ru-MOs) targeting two early developmental zebrafish genes, chordin and notail, were synthesized and tested in vivo. One-cell-stage zebrafish embryos microinjected with Ru-MO and incubated in the dark for 24 h developed normally, consistent with caging, whereas irradiation at 450 nm dissociated one 3-ethynylpyridine ligand (ϕ = 0.33) and uncaged the MO to achieve gene knockdown. As demonstrated, Ru photolinkers provide a versatile method for controlling structure and function of biopolymers.

Conformational studies of muscarone analogues: X-ray analysis and molecular mechanics calculations.

The X-ray structure of muscarone analogues 3 and 4 was determined and compared with that of muscarone (1, iodide and picrate salts), muscarine 2, dioxolane 5, oxathiolane 6, and tetrahydrofuran 7. In order to better define the pharmacological stereoselectivity of muscarone, the conformational profiles of compounds 1, 2, 3, and 5 were analyzed using Allinger's MM2(85) program or, in the case of 4, by 1H NMR spectroscopy. The conformation of the ring in 1 proved similar to that of the other derivatives. MM2 calculations predicted a preferred gauche arrangement of the side chain for 1 and its analogues; such an arrangement was also observed in the solid state of muscarone picrate. Thus, the antiperiplanar arrangement reported for crystalline muscarone iodide appears to be due to crystallographic packing forces. As a consequence, the rationalization of the pharmacological profile of 1 based on the antiperiplanar arrangement is now highly questionable. The lack of stereoselectivity of 4 can be attributed to the absence of a stereocenter at C-2 whereas, in our opinion, there are currently no sound explanations for the low values of eudismic ratios for the muscarone enantiomers.

The social hour for geropsychiatric patients.

This study was an attempt to evaluate the effects of a controlled alcohol intake in the socialization of a group of geropsychiatric patients in a clinical setting. Although 47 patients of the Geriatric Unit were involved at various times, 16 remained in the study for the entire eight weeks. The Social Hour consisted of a half-hour session every weekday. The alcohol served was whisky, gin, vodka, and wine. Soft drinks were available for those who preferred them. Each patient was rated on a 5-point scale with regard to ward and off-ward activities. A planned four-week placebo period met with so much negativism that it had to be shortened to three days. According to our observations, the socialization process was increased significantly for these mentally ill geriatric patients during this type of Social Hour.

Progress toward the total synthesis of callipeltin A (I): asymmetric synthesis of (3S,4R)-3,4-dimethylglutamine.

[reaction:see text] During the total synthesis of the novel cyclic depsipeptide callipeltin A (1), the unit (3S,4R)-3,4-dimethylglutamine, was successfully synthesized by asymmetric Michael addition and subsequent electrophilic azidation. The key feature of this approach is the generation of three adjacent stereogenic centers using the same camphorsultam chiral auxiliary.

Snapshots of titanium BINOLate complexes with diverse solid state structures.

[structure: see text]. Many important asymmetric reactions are catalyzed by (BINOLate)Ti species with unknown structures. Reported here are three structures of BINOLate titanium complexes that show an interesting aggregation of (BINOLate)Ti(OiPr)2 with itself and with titanium tetraisopropoxide. These complexes are potential intermediates in the asymmetric addition of alkyl groups to aldehydes.

Optimization of asymmetric catalysts using achiral ligands: metal geometry-induced ligand asymmetry.

[reaction: see text] Traditionally, asymmetric catalysts have been optimized by modification of resolved chiral ligands. In this Letter, we optimize the asymmetric addition of diethylzinc to aldehydes by modification of achiral methylene bis(phenol) ligands. Upon coordination of the substrate, the achiral ligand becomes asymmetric, a concept termed Metal Geometry-Induced Ligand Asymmetry. The enantioselectivity of the catalyst formed from a single resolved ligand and several achiral ligands ranged from 9% (R) to 83% (S).

Detection of entero- and verocyto-toxin genes in Escherichia coli from diarrhoeal disease in animals using the polymerase chain reaction.

Oligonucleotide primers were designed for the specific polymerase chain reaction (PCR) amplification of the enterotoxins STIa and LTI and of the verocytotoxins VT1 and VT2. All of 184 E. coli isolates from cases of diarrhoea from pigs, cattle and sheep gave identical toxin profiles by PCR and gene probe. Differentiation between VT2 and VT2v was achieved using two oligonucleotide primers pairs in PCR and showed that all of 34 VT2+ porcine isolates, of which 23 were 0138:K1, harboured VT2v whereas 20 VT2+ bovine and ovine isolates harboured VT2. No isolate harboured both VT2 polymorphs. Simplified methods for sample preparation for PCR were examined and showed that PCR was not inhibited by direct addition of broth culture to the reaction mixture.

Reality discrimination, reality monitoring and disposition towards hallucination.

This investigation tested the hypotheses that individuals disposed towards hallucination are deficient in the meta-cognitive skills of reality discrimination and reality monitoring, and that there is a strong correlation between the measures of these skills. Normal subjects scoring high or low on a scale measuring predisposition towards hallucination were tested on an auditory signal detection task. High scorers on the scale were found to differ from low scorers on a measure of perceptual bias but not on a measure of sensitivity. The same groups of subjects were tested on a reality monitoring measure in which they were required to monitor two different types of word: words they were required to listen to, and words they were required to listen to on some trials and imagine on others. On a surprise frequency test it was found that high scorers did not differ from low scorers on their ability to accurately count the memories of words they had heard. It was also found that both high scorers and low scorers overestimated the number of times they had heard words when they had also been required to imagine these words on tests; however, high scorers made this error to a significantly greater extent. A strong correlation was found between reality discrimination and reality monitoring measures for high scorers, but no correlation was found for low scorers.

Enhanced expression of 987P (F6) fimbrial adhesin by cultured enterotoxigenic Escherichia coli.

The expression of the 987P fimbrial adhesin by strains of enterotoxigenic Escherichia coli was enhanced and, in some cases, restored by passaging the organisms through Craigie's tubes. In contrast, the fimbrial adhesins K99, K88 and F41 were not optimally expressed in this medium. The results suggest that Craigie's tubes should be used for the optimum expression of 987P.

Experimental infection of neonatal pigs with CNF toxin-producing strains of Escherichia coli.

To examine and compare the pathogenicity of cytotoxic necrotising factor (CNF)-producing Escherichia coli, two litters of piglets were infected orally with 10(10) E coli O88 or 10(10) E coli O32 strains. Of the six piglets infected with E coli O88, two died within 24 hours and three developed blood-stained diarrhoea. The other piglets were killed one, five, six and eight days after infection, when bacterial cultures indicated an overwhelming bacteraemic infection with E coli O88 in the early stages followed by clearance through the large intestine. The pathological changes consisted of an early enteritis, progressing to enterocolitis and a bacteraemic spread to the lungs. The histopathological changes were characteristic of toxaemic effects in brain, heart, liver and kidney, and characterised by congestion, oedema and exudation. Infection with E coli O32 produced a milder but similar enterocolitis, also with bacterial colonisation in the lungs. The histopathological findings again reflected a toxaemia. The enteritis was more persistent after E coli O32 infection and the strain persisted in large numbers in the intestine. No evidence of bacterial adherence to the intestinal mucosa was found with either strain. Enteroinvasion was only evident in one E coli O88-infected piglet, but the consistent occurrence of interstitial pneumonia showed the predilection of these organisms for the lung. The results confirm the toxigenic properties of CNF+ E coli and suggest an important role for this organism in enteric infection of young pigs.

Escherichia coli isolated from farm animals in England and Wales between 1986 and 1991.

Escherichia coli isolated from farm animals in England and Wales between 1986 and 1991 were examined for their serogroup, fimbrial adhesin and toxin production, and resistance to antimicrobial drugs. Among the 3595 porcine isolates the more common serogroups were O149, O8, O138, O147 and O157. The F4(K88) adhesin was detected in 21 per cent of the cultures, whereas the other adhesins were never present in more than 3 per cent during any year, although there was an upward trend in the prevalence of the F6(987P) antigen. The heat labile enterotoxin was produced by more than 17 per cent of the cultures usually in association with the F4 antigen. The other toxins, heat stable (STa), verocytotoxin (VT) and cytotoxic necrotising factor (CNF) were produced by 5.3, 4.7 and 6.8 per cent of the cultures, respectively. Among the 1383 bovine cultures the more common serogroups were O8, O101 and O17. The F5(K99) adhesin was detected in 9.1 per cent of the cultures, usually in association with STa production. The toxins STa, VT and CNF were produced by 4.4, 2.8 and 8.3 per cent of the cultures, respectively. Among the 407 ovine isolates the more common serogroups were O8, O101 and O78, and 11 per cent of the cultures produced the F5(K99) antigen. The toxins STa, VT and CNF were produced by 1.5, 6.1 and 9.1 per cent of the cultures, respectively. In the case of the 402 poultry isolates the more common serogroups were O78, O2 and O8, which are usually associated with colisepticaemia. Resistance to antibiotics was common, particularly among the bovine isolates.

Diagnostic aspects of cerebrocortical necrosis.

Specimens from cattle and sheep suspected of having cerebrocortical necrosis (CCN) were studied. Rumenal contents were examined for thiaminase-producing bacteria. Thiaminase activity was assessed in rumenal contents. The thiamine concentration of liver, brain and heart was determined and erythrocyte transketolase assessed. Diagnosis in each case, whether positive or negative for CCN, was decided by histopathological examination. There was a substantial agreement between the biochemical findings and the histological diagnosis indicating that a provisional diagnosis may be made on clinical and biochemical data alone. The findings are discussed in relation to other diseases which have the same neuropathological features. Attempts to isolate thiaminase-producing bacteria, which may be implicated in the aetiology of CCN, were inconclusive.

A durable recording and feedback system.

A durable low-cost data-recording and feedback system is described. Data transcribed with a special pen onto a plastic sheet can be removed only with two solvents. These data can be displayed for feedback purposes without fear of destruction.