The synthesis of neurotensin antagonist SR 48692 for prostate cancer research.

An improved synthesis of the molecule SR 48692 is presented and its use as a neurotensin antagonist biological probe for use in cancer research is described. The preparation includes an number of enhanced chemical conversions and strategies to overcome some of the limiting synthetic transformations in the original chemical route.

Characterization of the chloroplast genome sequence of oil palm (Elaeis guineensis Jacq.).

Oil palm (Elaeis guineensis Jacq.) is an economically important crop, which is grown for oil production. To better understand the molecular basis of oil palm chloroplasts, we characterized the complete chloroplast (cp) genome sequence obtained from 454 pyrosequencing. The oil palm cp genome is 156,973 bp in length consisting of a large single-copy region of 85,192 bp flanked on each side by inverted repeats of 27,071 bp with a small single-copy region of 17,639 bp joining the repeats. The genome contains 112 unique genes: 79 protein-coding genes, 4 ribosomal RNA genes and 29 tRNA genes. By aligning the cp genome sequence with oil palm cDNA sequences, we observed 18 non-silent and 10 silent RNA editing events among 19 cp protein-coding genes. Creation of an initiation codon by RNA editing in rpl2 has been reported in several monocots and was also found in the oil palm cp genome. Fifty common chloroplast protein-coding genes from 33 plant taxa were used to construct ML and MP phylogenetic trees. Their topologies are similar and strongly support for the position of E. guineensis as the sister of closely related species Phoenix dactylifera in Arecaceae (palm families) of monocot subtrees.

Mapping cerebellar abiotrophy in Australian Kelpies.

An autosomal recessive form of cerebellar abiotrophy occurs in Australian Kelpie dogs. Clinical signs range from mild ataxia with intention tremor to severe ataxia with seizures. A whole-genome mapping analysis was performed using Affymetrix Canine SNP array v2 on 11 affected and 19 control dogs, but there was no significant association with disease. A homozygosity analysis identified a three megabase region likely to contain the disease mutation. The region spans 29.8-33 Mb on chromosome 3, for which all affected dogs were homozygous for a common haplotype. Microsatellite markers were developed in the candidate region for linkage analysis that resulted in a logarithm of odds score suggestive of linkage. The candidate region contains 29 genes, none of which are known to cause ataxia.

Sport-specific power assessment for rock climbing.

AIM: The popularity of rock climbing has resulted in a growing research base for the sport. However, at present there is a lack of sport-specific measures of performance in the field. The aim of this study was to examine the use of the powerslap test as a sport specific power measure. METHODS: The participants in this study were categorised into four different ability groups (novice, intermediate, advanced and elite) based on self reported lead grade. Two separate experiments were conducted to determine validity and reliability. The powerslap test was conducted on a revolution board with two variations - wide and narrow grip, for both sides of the body. The test started with the climber hanging at full extension from two holds from which a pull up movement was made releasing one hand to slap a scaled score board above. RESULTS: There was a significant relationship between powerslap scores and climbing ability (Left Wide: r=0.7, P<0.0005; right wide: r=0.69, P<0.0005; left narrow: r=0.73, P<0.0005; right narrow: r =0.72, P<0.0005). Further to this, scores on the powerslap narrow test were significantly differentiated by climber ability (LEFT: F(3,37)=15.74, P<0.0005; right: F(3,37)=12.16, P<0.0005). Limits of agreement and intra-class correlation indicated that the powerslap test is a reliable performance measure. CONCLUSION: According to the present findings the narrow grip variation of the powerslap test is a useful sport-specific power test that is related to climbing performance.

Splenic artery aneurysm presenting with clinical features of a bleeding gastric gastrointestinal stromal tumour.

Gastrointestinal stromal tumours often present with insidious upper gastrointestinal symptoms. Initial definitive diagnosis can be difficult and therefore misdiagnosis is not infrequent. Here we report a case of upper GI bleeding caused by a splenic artery aneurysm that was misdiagnosed as a gastric GIST. This rare presentation of splenic artery aneurysm highlights the potential pitfalls of investigation in upper gastrointestinal disease.

The multifaceted proteins MvaT and MvaU, members of the H-NS family, control arginine metabolism, pyocyanin synthesis, and prophage activation in Pseudomonas aeruginosa PAO1.

The MvaT and MvaU proteins belonging to the H-NS family were identified as DNA-binding proteins that interact with the regulatory region of the aotJQMOP-argR operon for arginine uptake and regulation. Recombinant MvaT and MvaU proteins were purified, and binding of these purified proteins to the aotJ regulatory region was demonstrated using electromobility shift assays. Polyclonal antibodies against purified MvaT and MvaU were prepared and employed in supershift assays to support these observations. Knockout mutations resulting in a single lesion in mvaT or mvaU, as well as knockout mutations resulting in double lesions, were constructed using biparental conjugation, and the absence of MvaT and MvaU in the resulting mutants was confirmed by immunoblot analysis. Using measurements of the beta-galactosidase activities from aotJ::lacZ fusions in the mutants and the parental strain, it was found that MvaT and MvaU serve as repressors in control of aotJ expression. The effects of MvaT and MvaU on pyocyanin synthesis and CupA fimbrial expression in these mutants were also analyzed. Pyocyanin synthesis was induced in the single mutants but was completely abolished in the double mutant, suggesting that there is a complicated regulatory scheme in which MvaT and MvaU are essential elements. In comparison, MvaT had a more profound role than MvaU as a repressor of cupA expression; however, a combination of MvaT depletion and MvaU depletion had a strong synergistic effect on cupA. Moreover, prophage Pf4 integrated into the chromosome of Pseudomonas aeruginosa PAO1 was activated in an mvaT mvaU double mutant but not in a single mutant. These results were supported by purification and nucleotide sequencing of replicative-form DNA and by the release of phage particles in plaque assays. In summary, the mvaT mvaU double mutant was viable, and depletion of MvaT and MvaU had serious effects on a variety of physiological functions in P. aeruginosa.

Changes in physiological parameters in overtrained Standardbred racehorses.

Various changes in physiological parameters are associated with overtraining, which can be a serious problem for human and equine athletes. A 34 week longitudinal study was conducted to investigate the effects of an acute training overload on physiological parameters in 10 Standardbred racehorses. After 24 weeks of training, horses received 8 weeks of increased workload, followed by 2 weeks recovery. Horses performed a 2400 m time trial and a progressive submaximal exercise test on alternate weeks. By the end of the heavy training period, the average time for the final 1200 m of the time trial increased by 4.0% (95% probable range of true value 1.7-5.8) and peak velocity decreased by 6.9% (4.7-8.9), indicating that overtraining had occurred. Acute overtraining coincided with an increase in blood lactate concentration after the time trial and submaximal test. There were also substantial decreases in bodyweight, plasma cortisol concentration and packed cell volume after the time trial, and in the velocity at a heart rate of 200/min (V200). Parameters that showed no clear-cut change with overtraining included maximal and recovery heart rate, basal plasma cortisol, plasma and red cell volume, and markers of skeletal damage (plasma concentrations of creatine kinase and aspartate aminotransferase). Bodyweight, V200, postexercise blood lactate and plasma cortisol concentrations may all be useful for detecting acute overtraining in equine athletes.

Effect of tapered normal and interval training on performance of Standardbred pacers.

Human athletes taper or reduce their training load before a race to enhance performance, apparently because recovery from the effects of fatigue occurs faster than the loss of fitness from the reduced training. However, there appear to be no previous studies of tapering of equine athletes. Our aim in the present study was, therefore, to investigate the efficacy of tapering with Standardbred pacers. We determined the effect of repeated cycles of tapered training on performance of Standardbred pacers. After 8 weeks of jogging and 3 x 2 week cycles of pace work, 19 horses were randomised to a taper and a control group. The taper group completed 5 consecutive 2 week cycles, each incorporating a 7 day taper; some cycles included high-intensity interval training. The control group continued with 5 more cycles of pace work. All horses completed a 2400 m individual time trial after each cycle. Peak and mean speed of the taper group were faster than those of the control group in all cycles; the differences were clear-cut in all cycles for peak speed (overall 4.4%, 95% confidence interval 1.7 to 7.1%), but only in one of the interval-training cycles for mean speed (2.4%, 0.3 to 4.7%). Four horses in the taper group were injured during interval training. Repeated tapering produces a worthwhile enhancement of performance in Standardbreds, but the addition of interval training appears to increase the risk of injury.

Reliability of two 2,400-m time-trial protocols for assessing performance of Standardbred racehorses.

OBJECTIVE: To evaluate the reliability of 2 time-trial protocols, 1 that involved use of a pacemaker and 1 that did not, for assessing performance of Standardbred racehorses. ANIMALS: 3- to 7-year-old Standardbred maiden pacers. PROCEDURE: 11 Standardbred pacers were used to determine the reliability of a time-trial protocol that involved use of a galloping pacemaker. All tests were performed by a single driver. Horses were paced for an initial 1,600 m then raced and timed for the final 800 m. They were retested 9 days later. Twenty-two horses were used to determine the reliability of a time-trial protocol that did not involve use of a pacemaker. All tests were performed by a second driver. Horses were paced for the first 1,200 m then raced and timed for the final 1,200 m. They were retested 3 and 8 days later. RESULTS: Reliability of performance time, expressed as the coefficient of variation (ie, the typical percentage variation in a horse's time between trials), was similar for the 2 protocols (1.0% and 1.3%, respectively). CONCLUSIONS: The small variations in performance times for the 2 time-trial protocols were similar to those of the best comparable laboratory and field tests of human and equine performance. Both protocols would be suitable for investigating factors affecting performance of Standardbred racehorses.

A retrospective anonymous pilot study in screening newborns for HFE mutations in Scandinavian populations.

We have retrospectively analyzed 837 random anonymized dried blood spot (DBS) samples from neonatal screening programs in Scandinavia for mutations in HFE, the candidate gene for hemochromatosis. We have found C282Y allele frequencies of 2.3% (+2.0%) (-1.3%) in Greenland, 4.5%+/-1.9% in Iceland, 5.1%+/-2.3% in the Faeroe Islands, and 8.2%+/-2.7% in Denmark. The high prevalence of HFE mutations in Denmark suggests that population screening for the C282Y mutation could be highly advantageous in terms of preventive health care. Long-term follow-up evaluation of C282Y homozygotes and H63D/C282Y compound heterozygotes will give an indication of the penetrance of the mutations.

The effect of HFE mutations on serum ferritin and transferrin saturation in the Jersey population.

High frequencies of the haemochromatosis-related HFE C282Y mutation have been reported in North European populations, in which a high proportion of patients with the disease are homozygotes. However, the degree of penetrance of this genotype is unknown. We determined the HFE C282Y and H63D genotypes of 411 consenting volunteer blood donors on Jersey, and the serum ferritin and transferrin saturation levels of 204 of these volunteers. The C282Y allele frequency was found to be 8.3% in 822 chromosomes, indicating a homozygote frequency of 1/145. Consistent with this, four C282Y homozygotes were detected in 411 volunteers. As there are only 18 patients presently receiving treatment for haemochromatosis on Jersey, out of a total population of about 85000, there is a large discrepancy between the number of haemochromatosis patients and the number of C282Y homozygotes in this population. In a preliminary study of 204 consenting volunteers we found a correlation between transferrin saturation and HFE H63D/ C282Y genotype (P=0.017) and between serum ferritin and genotype (P = 0.056). We also observed elevated values of transferrin saturation in the two C282Y homozygotes assayed. These results suggest that a large proportion of the many undetected C282Y homozygotes on Jersey and in similar populations could be in the preclinical stages of haemochromatosis, and warrant investigation. However, there may be a wide variation in the expression of the condition, and a more extensive study of the level of disease penetrance encompassing a large number of hitherto undetected C282Y homozygotes is therefore imperative.

Rapid mapping of markers applying vectorette technology to YAC fragmentation allows easy assembly of a high-density STS bacterial clone contig spanning the markers D6S1260-D6S1918.

We have generated a detailed physical map of the 6p21.3/p22.1 boundary, using a combination of yeast artificial chromosome (YAC) fragmentation and high-resolution sequence tagged site (STS) content mapping. YACs from the CEPH, St. Louis, and ICRF libraries have been used to construct a 4.5-Mb contig spanning the markers D6S306 to D6S1571. YAC insert sizes were determined by pulsed field gel electrophoresis (PFGE). Chimerism of YACs was determined by fluorescent in situ hybridization (FISH), and their integrity was determined by fingerprinting with Alu-PCR. We have identified 10 new CA repeat loci in this region as well as over 50 novel STSs, several tRNA genes, a new histone H2B gene and the phospholipase D gene. Using these new markers, we have rapidly generated a bacterial clone contig of over 250 kb, spanning the markers D6S1260 to D6S1918 (WI-3111) with STSs spaced on average every 6 kb.

The HLA A1-B8 haplotype extends 6 Mb beyond HLA-A: associations between HLA-A, B, F and 15 microsatellite markers.

Allele frequencies for HLA-A, B and F and 15 microsatellite markers located from 100 kb telomeric to HLA-A to 6 Mb telomeric have been determined in a group of 60 blood donors. Linkage disequilibrium analysis revealed significant haplotype associations even after correction for the number of comparisons made. The HLA-A1, B8 haplotype extends as far as D6S276 (6.0 Mb telomeric to HLA-A). It is important to realize that this common haplotype extends beyond the HLA region, especially when evaluating haplotype associations with particular disorders.

A rapid non-invasive method for the detection of the haemochromatosis C282Y mutation.

We describe the rapid single-step detection, by mutagenically separated polymerase chain reaction (MS-PCR), of the HLE C282Y mutation, for which > 90% of haemochromatosis patients in the U.K. are homozygous. In addition to using purified DNA as a template, whole blood and lysed buccal cell extracts from mouthwash samples can be used. Therefore sample collection may be non-invasive and purification steps kept to a minimum.