Growth restriction in the rat alters expression of cardiac JAK/STAT genes in a sex-specific manner.

Uteroplacental insufficiency resulting in intrauterine growth restriction has been associated with the development of cardiovascular disease, coronary heart disease and increased blood pressure, particularly in males. The molecular mechanisms that result in the programming of these phenotypes are not clear. This study investigated the expression of cardiac JAK/STAT signalling genes in growth restricted offspring born small due to uteroplacental insufficiency. Bilateral uterine vessel ligation was performed on day 18 of pregnancy to induce growth restriction (Restricted) or sham surgery (Control). Cardiac tissue at embryonic day (E) 20, postnatal day (PN) 1, PN7 and PN35 in male and female Wistar (WKY) rats (n=7-10 per group per age) was isolated and mRNA extracted. In the heart, there was an effect of age for males for all genes examined there was a decrease in expression after PN1. With females, JAK2 expression was significantly reduced after E20, while PI3K in females was increased at E30 and PN35. Further, mRNA expression was significantly altered in JAK/STAT signalling targets in Restricteds in a sex-specific manner. Compared with Controls, in males, JAK2 and STAT3 were significantly reduced in the Restricted, while in females SOCS3 was significantly increased and PI3K significantly decreased in the Restricted offspring. Finally, there were specific differences in the levels of gene expression within the JAK/STAT pathway when comparing males to females. Thus, growth restriction alters specific targets in the JAK/STAT signalling pathway, with altered JAK2 and STAT3 potentially contributing to the increased risk of cardiovascular disease in the growth restricted males.

beta-1,3-Glucuronyltransferase-1 gene implicated as a candidate for a schizophrenia-like psychosis through molecular analysis of a balanced translocation.

We have mapped and sequenced both chromosome breakpoints of a balanced t(6;11)(q14.2;q25) chromosome translocation that segregates with a schizophrenia-like psychosis. Bioinformatics analysis of the regions revealed a number of confirmed and predicted transcripts. No confirmed transcripts are disrupted by either breakpoint. The chromosome 6 breakpoint region is gene poor, the closest transcript being the serotonin receptor 1E (HTR1E) at 625 kb telomeric to the breakpoint. The chromosome 11 breakpoint is situated close to the telomere. The closest gene, beta-1,3-glucuronyltransferase (B3GAT1 or GlcAT-P), is 299 kb centromeric to the breakpoint. B3GAT1 is the key enzyme during the biosynthesis of the carbohydrate epitope HNK-1, which is present on a number of cell adhesion molecules important in neurodevelopment. Mice deleted for the B3GAT1 gene show defects in hippocampal long-term potentiation and in spatial memory formation. We propose that the translocation causes a positional effect on B3GAT1, affecting expression levels and making it a plausible candidate for the psychosis found in this family. More generally, regions close to telomeres are highly polymorphic in both sequence and length in the general population and several studies have implicated subtelomeric deletions as a common cause of idiopathic mental retardation. This leads us to the hypothesis that polymorphic or other variation of the 11q telomere may affect the activity of B3GAT1 and be a risk factor for schizophrenia and related psychoses in the general population.

Phylogenetic analysis of coccidia based on 18S rDNA sequence comparison indicates that Isospora is most closely related to Toxoplasma and Neospora.

The phylogenetic relationships and taxonomic affinities of coccidia with isosporan-type oocysts have been unclear as overlapping characters, recently discovered life cycle features, and even recently discovered taxa, continue to be incorporated into biological classifications of the group. We determined the full or partial 18S ribosomal RNA gene sequences of three mammalian Isospora spp., Isospora felis, Isospora ohioensis and Isospora suis, and a Sarcocystis sp. of a rattlesnake, and used these sequences for a phylogenetic analysis of the genus Isospora and the cyst-forming coccidia. Various alveolate 18S rDNA sequences were aligned and analyzed using maximum parsimony to obtain a phylogenetic hypothesis for the group. The three Isospora spp. were found to be most closely related to Toxoplasma gondii and Neospora caninum. This clade in turn formed the sister group to the Sarcocystis spp. included in the analysis. The results confirm that the genus Isospora does not belong to the family Eimeriidae, but should be classified together with the cyst-forming coccidia in the family Sarcocystidae. Furthermore, there appear to be two lineages within the Sarcocystidae. One lineage comprises Isospora and the Toxoplasma/Neospora clade which share the characters of having a proliferative phase of development preceding gamogony in the definitive host and an exogenous phase of sporogony. The other lineage comprises the Sarcocystis spp. which have no proliferative phase in the definitive host and an endogenous phase of sporogony.

Identification of synapomorphic characters in the genus Sarcocystis based on 18S rDNA sequence comparison.

In order to further investigate synapomorphic characters in the genus Sarcocystis, the small subunit ribosomal RNA gene sequences of Sarcocystis capracanis and Sarcocystis moulei were determined and used to infer the phylogenetic position of these two organisms within the cyst-forming coccidia. Phylogenies derived using distance, maximum parsimony and maximum likelihood methods demonstrated that S. capracanis groups with Sarcocystis tenella and Sarcocystis arieticanis as a clade that shares the characteristic of using canids as their definitive host. S. moulei was shown to group with Sarcocystis gigantea and Sarcocystis fusiformis as a clade that shares the characteristic of using fields as their definitive host.

Genetic diversity in Sarcocystis gigantea assessed by RFLP analysis of the ITS1 region.

The genetic diversity among Sarcocystis gigantea isolates derived from individual cysts within a given infected animal at two abattoirs in Australia and one abattoir in Germany was studied using Restriction Fragment Length Polymorphism (RFLP) analysis of the intergenic transcribed spacer region 1 (ITS 1) of the ribosomal RNA gene operon. S. gigantea isolates were obtained from infected sheep from Blayney (New South Wales), Katanning (Western Australia), and Detmold (North-Rhine Westfalia, Germany) in order to assess the level of diversity among isolates from different geographic locations. Polymerase chain reaction amplification and RFLP analysis of the ITSI region with the restriction enzymes HaeIII, NlaIII, and Sau3AI found no genetic variation among the isolates within one animal or among animals in the same or different locations. To our knowledge, such a field study has not yet been performed on a species in the genus Sarcocystis.

A RAPD-PCR derived marker can differentiate between pathogenic and non-pathogenic Sarcocystis species of sheep.

Random amplified polymorphic DNA (RAPD)-PCR was used to differentiate among four cyst-forming coccidia of sheep, Sarcocystis tenella, Sarcocystis gigantea, Sarcocystis arieticanis, and Toxoplasma gondii. Genomic DNA of the four parasite species was amplified using RAPD-PCR and the DNA fragments were separated on agarose gels. A RAPD-PCR band derived from S. tenella was isolated from the gel and subcloned into pUC18. The insert was sequenced and found to be 1278 nucleotides long. This sequence appeared to be cryptic in nature as it showed no significant sequence peculiarities or similarity with any other known sequences either at the nucleotide or derived amino acid levels. The recombinant plasmid was radiolabelled and used as a probe in Southern hybridization. This probe, termed pSTF10, hybridised to Mbo I restricted genomic DNA of S. tenella and S. arieticanis, but not to DNA of S. gigantea, T. gondii, mouse, or sheep. It is likely that STF10 will become a valuable diagnostic tool for Sarcocystis infections in sheep to differentiate between pathogenic species of this genus and S. gigantea or T. gondii.

Wear patterns of the articular cartilage and triangular fibrocartilaginous complex of the wrist: a cadaveric study.

The incidence and patterns of degenerative changes within the radio-carpal joint were studied in 138 specimens of elderly cadaveric wrists. Articular cartilage wear of varying severity was seen on the distal radial and ulnar articular surfaces in 27% of cases and on the proximal row articular surfaces in 54%. Wear was most commonly seen on the radial styloid and corresponding area of the scaphoid. The triangular fibrocartilaginous complex (TFCC) was found to be degenerate or torn in 24%. Central degenerative perforation was commonly associated with articular cartilage wear on the ulnar head and the ulnar half of the lunate. No significant wear pattern was seen in those wrists with peripheral linear (i.e. traumatic) TFCC tears. Interosseous scapho-lunate and luno-triquetral ligament disruptions were found in less than 10%, suggesting that disruption of these ligaments is usually traumatic and not degenerative.

Radiation dose rates from adult patients undergoing nuclear medicine investigations.

Adult patients undergoing nuclear medicine investigations may subsequently come into close contact with members of the public and hospital staff. In order to expand the available dosimetry and derive appropriate recommendations, dose rates were measured at 0.1, 0.5 and 1.0 m from 80 adult patients just before they left the nuclear medicine department after undergoing one of eight 99Tcm studies, an 123I thyroid, an 111In leucocyte or a 201Tl cardiac scan. The maximum departure dose rates at these distances of 150, 30 and 7.3 microSv h-1 were greater than those found in similar published studies of adult and paediatric patients. To limit the dose to an infant to less than 1 mSv, an 111In leucocyte scan is the only investigation for which it may be necessary to restrict close contact between the infant and a radioactive parent, depending on the dose rate near the surface of the patient, the parent's habits and how fretful is the infant. It is unlikely that a ward nurse will receive a dose of 60 microSv in a working day if caring for just one radioactive adult patient, unless the patient is classified as totally helpless and has undergone a 99Tcm marrow, bone or brain scan. The data and revised calculations of effective exposure times based on a total close contact time of 9 h in every 24 h period should allow worst case estimates of radiation dose to be made and recommendations to be formulated for other circumstances, including any future legislative changes in dose limits or derived levels.

An optimized assay for adenosine deaminase using reverse phase high pressure liquid chromatography.

A sensitive, optimized assay for adenosine deaminase (E.C. 3.5.4.4), is presented which is based on a reverse phase HPLC analysis of adenosine. In this method a sample of erythrocytes was incubated with adenosine and the decrease in the adenosine concentration with time was analyzed by HPLC. The precision of the method averaged approximately 5% RSD with a sensitivity of about 0.1 U/ml of packed erythrocytes. Comparison with other literature values showed similar activities for adenosine deaminase in erythrocytes (0.229 +/- 0.025) U/ml (alpha = 0.05). The optimization included studies on the ionic strength, pH, enzyme and substrate concentration, and reaction time. The Km for adenosine deaminase was found to be (0.178 +/- 0.018) mM (alpha = 0.05). The method offers several advantages over other assay methods, including an improved capability to discern competing side reactions from other enzymes.