An RFLP for probe 26-6 that is useful in linkage diagnosis for adult polycystic kidney disease

A previously unreported RFLP for probe 26-6, at 1.2-kb, has been recorded in two families after Southern blotting, where it has been useful in establishing phase. This allele is not detected in the polymerase chain reaction (PCR) assay for 26-6 where individuals are typed only as E₂.     

Forest-scale sap flux responses to rainfall in a dryland eucalyptus plantation

Dryland salinity is caused by rising saline water tables, the result of relatively recent landscape-scale clearance of deep-rooted vegetation. One obvious solution to this problem is the reintroduction of deep-rooted vegetation into these landscapes, most likely non-deciduous trees. Ideally, continually-transpiring deep-rooted trees would remove moisture from throughout the soil profile, increasing the capacity of the soil to store water, thus lowering water tables by effectively reducing the number of rainfall events that contribute to groundwater recharge. In this study, we examined how water use by a Eucalyptus sideroxylon A. Cunn. ex Woolls plantation, growing in a salinity-prone landscape, varied in response to rainfall events across four years of sap flux monitoring. Responses of the plantation were observed across multiple seasons, from above average to well below average rainfall. We observed that the plantation forest, while capable of continuous water use during drought, was also quite responsive to rainfall events. During the driest periods, during which shallow soil moisture was reduced to a stable minimum, the forest continued using water at around 1 mm/day. Generally we observed increases in forest water use following only 5 mm of rainfall, in contrast to 20 mm for neighbouring native vegetation. We compared a range of plausible empirical models for describing forest water use responses to rainfall. The best model demonstrated that rainfall size, post-rainfall PET and the interaction between rainfall size and antecedent soil moisture made significant contributions to variation in forest water use across rainfall events. Interestingly, the model showed that all else equal, higher antecedent soil moisture tended to reduce potential increases in forest water use in response to rainfall.     

Attenuation of radiation-induced genomic instability by free radical scavengers and cellular proliferation.

To investigate the mechanisms of radiation-induced chromosomal instability, cells were irradiated in the presence of the free radical scavengers DMSO, glycerol, or cysteamine, in the presence of DMSO while frozen, or held in confluence arrest post-irradiation to permit cells to repair potentially lethal DNA damage. Clones derived from single progenitor cells surviving each treatment were then analyzed for the subsequent development of chromosomal instability. The presence of scavengers (+/- freezing) during irradiation, and the recovery from potentially lethal damage after irradiation led to an increase in cell survival that was accompanied by a decrease in the initial yield of chromosomal rearrangements. Furthermore, analysis of over 400 clones and 80,000 metaphases indicates that these same treatments reduced the incidence of instability at equitoxic doses when compared to controls irradiated in the absence of scavengers at ambient temperature. Results suggest that preventing reactive species from damaging DNA, promoting chemical repair of ionized DNA intermediates, or allowing enzymatic removal of genetic lesions, represent measures that reduce the total burden of DNA damage and reduce the subsequent onset of radiation-induced genomic instability.     

Phylogenetic analysis of 1.5 Mbp and platypus EST data refute the Marsupionta hypothesis and unequivocally support Monotremata as sister group to Marsupialia/Placentalia

The extant mammalian groups Monotremata, Marsupialia and Placentalia are, according to the 'Theria' hypothesis, traditionally classified into two subclasses. The subclass Prototheria includes the monotremes and subclass Theria marsupials and placental mammals. Based on some morphological and molecular data, an alternative proposition, the Marsupionta hypothesis, favours a sister group relationship between monotremes and marsupials to the exclusion of placental mammals. Phylogenetic analyses of single genes and even multiple gene alignments have not yet been able to conclusively resolve this basal mammalian divergence. We have examined this problem using one data set composed of expressed sequence tags (EST) and another containing 1 510 509 nucleotide (nt) sites from 1358 inferred cDNA genomic sequences. All analyses of the concatenated sequences unambiguously supported the Theria hypothesis. The Marsupionta hypothesis was rejected with high statistical confidence from both data sets. In spite of the strong support for Theria, a non-negligible number of single genes supported either of the two alternative hypotheses. The divergence between monotremes and therian mammals was estimated to have taken place 168–178 Mya, a dating compatible with the fossil record. Considering the long common evolutionary branch of therians, it is surprising that sequence data from many thousand amino acid sites were needed to conclusively resolve their relationship to monotremes. This finding draws attention to other mammalian divergences that have been taken as unequivocally settled based on much smaller alignments. EST data provide a comprehensive random sample of protein coding sequences and an economic way to produce large amounts of data for phylogenetic analysis of species for which genomic sequences are not yet available.     

An unreported RFLP for probe 218 EP6 that is useful in linkage analysis of adult polycystic kidney disease

Probe 218 EP6 is known to recognise two restriction fragment length polymorphisms (RFLPs) after digestion of genomic DNA withPvuII. We report a rare allele that segregates in Mendelian fashion, in a family where adult polycystic kidney disease was being tracked using linked polymorphisms.     

Development of an Automated and Sensitive Microfluidic Device for Capturing and Characterizing Circulating Tumor Cells (CTCs) from Clinical Blood Samples

Current analysis of circulating tumor cells (CTCs) is hindered by sub-optimal sensitivity and specificity of devices or assays as well as lack of capability of characterization of CTCs with clinical biomarkers. Here, we validate a novel technology to enrich and characterize CTCs from blood samples of patients with metastatic breast, prostate and colorectal cancers using a microfluidic chip which is processed by using an automated staining and scanning system from sample preparation to image processing. The Celsee system allowed for the detection of CTCs with apparent high sensitivity and specificity (94% sensitivity and 100% specificity). Moreover, the system facilitated rapid capture of CTCs from blood samples and also allowed for downstream characterization of the captured cells by immunohistochemistry, DNA and mRNA fluorescence in-situ hybridization (FISH). In a subset of patients with prostate cancer we compared the technology with a FDA-approved CTC device, CellSearch and found a higher degree of sensitivity with the Celsee instrument. In conclusion, the integrated Celsee system represents a promising CTC technology for enumeration and molecular characterization.