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Acute kidney injury (AKI) is a global clinical problem characterised by a sudden decline in renal function and mortality as high as 60%. Current AKI biomarkers have limited ability to classify disease progression and identify underlying pathological mechanisms. Here we hypothesised that alterations in urinary microRNA profiles could predict AKI recovery/nonrecovery after 90 days, and that injury-specific changes would signify microRNA mediators of AKI pathology. Comparison of urinary microRNA profiles from AKI patients with controls detected significant injury-specific increases in miR-21, miR-126 and miR-141 (p . Forced miR-141 expression in the presence of H2O2 increased PTEC death and decreased cell viability. Of nine messenger RNA targets with two or more miR-141 3’-untranslated region binding sites, we confirmed protein tyrosine phosphatase receptor type G (PTPRG) as a direct miR-141 target in PTECs. PTPRG-specific siRNA knockdown under oxidative stress increased PTEC death and decreased cell viability. In conclusion, we detected significant alterations in five urinary microRNAs following AKI, and identified proximal tubular cell PTPRG as a putative novel therapeutic target.

This paper describes a straightforward electrochemical method for rapid and robust urinary microRNA (miRNA) quantification using disposable biosensors that can discriminate between urine from diabetic kidney disease (DKD) patients and control subjects. Aberrant miRNA expression has been observed in several major human disorders, and we have identified a urinary miRNA signature for DKD. MiRNAs therefore have considerable promise as disease biomarkers, and techniques to quantify these transcripts from clinical samples have significant clinical and commercial potential. Current RT-qPCR-based methods require technical expertise, and more straightforward methods such as electrochemical detection offer attractive alternatives. We describe a method to detect urinary miRNAs using diazo sulfonamide-modified screen printed carbon electrode-based biosensors that is amenable to parallel analysis. These sensors showed a linear response to buffered miR-21, with a 17 fM limit of detection, and successfully discriminated between urine samples (n = 6) from DKD patients and unaffected control subjects (n = 6) by differential miR-192 detection. Our technique for quantitative miRNA detection in liquid biopsies has potential for development as a platform for non-invasive high-throughput screening and/or to complement existing diagnostic procedures in disorders such as DKD., In this study we have developed an electrochemical microRNA biosensor sensitive to 17 fM and capable of detecting an established downregulation of urinary miR-192 in diabetic kidney disease patients.

The Arabidopsis thaliana genome encodes a family of 28 proteins whose members have been associated with the transport of monovalent cations across membranes. Experiments have been performed to elucidate the biochemical function and the role in plant development of two closely related members of this CHX family. A genotype carrying a knockout of the AtCHX23 gene (At1g05580) showed no phenotype when grown in glasshouse conditions. In particular, it did not exhibit the reduced root growth phenotype observed for a knockout of its homologue AtCHX21 when exposed to elevated sodium concentration. However, it was not possible to produce plants that were homozygous knockout for both AtCHX21 and AtCHX23. Reverse transcription-PCR (RT-PCR) experiments revealed that both genes are highly expressed in flower buds, flowers, and pollen. However, examination of pollen grain viability and pollen tube growth through excised styles did not reveal a phenotypic difference between the chx21–chx23– condition and other haplotypes. Crosses between selected mutants and wild-type plants in which the chx21–chx23– haplotype was produced by either the male or female parent demonstrated unequivocally that the chx21–chx23– haplotype could not pass through the female line. This suggests that the genes share a critical function in the development and/or function of the female gametophyte and that this function cannot be provided by other members of the AtCHX gene family. Experiments were carried out using the heterologous expression of AtCHX23 in Saccharomyces cerevisiae genotypes carrying combinations of deletions of genes involved in the transport of sodium or potassium across membranes. The results show that CHX23 would only complement the poor colony growth phenotype associated with the deletion of the yeast gene kha1. The conclusion is that both AtCHX21 and AtCHX23 act in potassium homeostasis within the female gametophyte and this is discussed in terms of the diversification of gene sequence and function within the CHX gene family.

Urinary microRNAs show promise as noninvasive biomarkers in renal disease. Here, we describe a detailed protocol for the column-based extraction and quantification of miRNAs by reverse-transcription quantitative polymerase chain reaction (RT-qPCR) from urine samples.

Background Information is scarce regarding effects of antihypertensive medication on blood pressure variability ( BPV ) and associated clinical outcomes. We examined whether antihypertensive treatment changes BPV over time and whether such change (decline or increase) has any association with long‐term mortality in an elderly hypertensive population. Methods and Results We used data from a subset of participants in the Second Australian National Blood Pressure study (n=496) aged ≥65 years who had 24‐hour ambulatory blood pressure recordings at study entry (baseline) and then after a median of 2 years while on treatment (follow‐up). Weighted day‐night systolic BPV was calculated for both baseline and follow‐up as a weighted mean of daytime and nighttime blood pressure standard deviations. The annual rate of change in BPV over time was calculated from these BPV estimates. Furthermore, we classified both BPV estimates as high and low based on the baseline median BPV value and then classified BPV changes into stable: low BPV , stable: high BPV , decline: high to low , and increase: low to high . We observed an annual decline (mean± SD : −0.37±1.95; 95% CI, −0.54 to −0.19; P BPV between baseline and follow‐up. Having constant stable: high BPV was associated with an increase in all‐cause mortality (hazard ratio: 3.03; 95% CI, 1.67–5.52) and cardiovascular mortality (hazard ratio: 3.70; 95% CI, 1.62–8.47) in relation to the stable: low BPV group over a median 8.6 years after the follow‐up ambulatory blood pressure monitoring. Similarly, higher risk was observed in the decline: high to low group. Conclusions Our results demonstrate that in elderly hypertensive patients, average BPV declined over 2 years of follow‐up after initiation of antihypertensive therapy, and having higher BPV (regardless of any change) was associated with increased long‐term mortality.

Chronic Kidney Disease is a growing problem across the world and can lead to end-stage kidney disease and cardiovascular disease. Fibrosis is the underlying mechanism that leads to organ dysfunction, but as yet we have no therapeutics that can influence this process. Ras monomeric GTPases are master regulators that direct many of the cytokines known to drive fibrosis to downstream effector cascades. We have previously shown that K-Ras is a key isoform that drives fibrosis in the kidney. Here we demonstrate that K-Ras expression and activation are increased in rodent models of CKD. By knocking down expression of K-Ras using antisense oligonucleotides in a mouse model of chronic folic acid nephropathy we can reduce fibrosis by 50% and prevent the loss of renal function over 3 months. In addition, we have demonstrated in vitro and in vivo that reduction of K-Ras expression is associated with a reduction in Jag1 expression; we hypothesise this is the mechanism by which targeting K-Ras has therapeutic benefit. In conclusion, targeting K-Ras expression with antisense oligonucleotides in a mouse model of CKD prevents fibrosis and protects against renal dysfunction.

Predicting immediate and subsequent graft function is important in clinical decision-making around kidney transplantation, but is difficult using available approaches. Here we have evaluated urinary microRNAs as biomarkers in this context. Profiling of 377 microRNAs in the first urine passed post-transplantation identified 6 microRNAs, confirmed to be upregulated by RT-qPCR in an expanded cohort (miR-9, -10a, -21, -29a, -221, and -429, n = 33, P

A pressing need for new chronic kidney disease (CKD) biomarkers persists. MicroRNAs (miRNAs) are emerging as a novel class of disease biomarkers in body fluids, but mechanisms conferring their stability in urine have not been fully elucidated. Here we investigated stabilization in human urine of ubiquitously expressed miR-16, and miR-192, which we have shown previously to be downregulated in renal fibrosis, by association with extracellular vesicles and with argonaute protein (AGO) 2. Endogenous urinary miR-16 was significantly more resistant to RNase-mediated degradation than exogenous, spiked-in, Caenorhabditis elegans cel-miR-39. We used our previously optimized high-resolution exosome isolation protocol with sucrose gradient ultracentrifugation to sub-fractionate the primary extracellular vesicle-rich urinary pellet. MiR-16 and miR-192 were enriched in exosomal sucrose gradient fractions, but were also detected in all other fractions. This suggested association of urinary miRNAs with other urinary extracellular vesicles and/or pellet components, complicating previous estimates of miRNA:exosome stoichiometry. Proteinase K digestion destabilized urinary miR-16 and we showed, for the first time, RNA-immunoprecipitation of urinary miR-16:AGO2 and miR-192:AGO2 complexes. Association with exosomes and AGO2 stabilized urinary miR-16 and miR-192, suggesting quantitative urinary miRNA analysis has the potential to identify novel, non-invasive CKD biomarkers.

Hyaluronan (HA) is a ubiquitous extracellular matrix glycosaminoglycan composed of repeated disaccharide units of alternating D-glucuronic acid and D-N-acetylglucosamine residues linked via alternatingβ-1,4 andβ-1,3 glycosidic bonds. HA is synthesized in humans by HA synthase (HAS) enzymes 1, 2, and 3, which are encoded by the correspondingHASgenes. Previous in vitro studies have shown characteristic changes in HAS expression and increased HA synthesis in response to wounding and proinflammatory cytokines in human peritoneal mesothelial cells. In addition, in vivo models and human peritoneal biopsy samples have provided evidence of changes in HA metabolism in the fibrosis that at present accompanies peritoneal dialysis treatment. This review discusses these published observations and how they might contribute to improvement in peritoneal dialysis.

Effective diabetic kidney disease (DKD) biomarkers remain elusive, and urinary miRNAs represent a potential source of novel noninvasive disease sentinels. We profiled 754 miRNAs in pooled urine samples from DKD patients (n = 20), detecting significantly increased miR-126, miR-155, and miR-29b compared with controls (n = 20). These results were confirmed in an independent cohort of 89 DKD patients, 62 diabetic patients without DKD, and 41 controls: miR-126 (2.8-fold increase; P < 0.0001), miR-155 (1.8-fold increase; P < 0.001), and miR-29b (4.6-fold increase; P = 0.024). Combined receiver operating characteristic curve analysis resulted in an area under the curve of 0.8. A relative quantification threshold equivalent to 80% sensitivity for each miRNA gave a positive signal for 48% of DKD patients compared with 3.6% of diabetic patients without DKD. Laser-capture microdissection of renal biopsy specimens, followed by quantitative RT-PCR, detected miR-155 in glomeruli and proximal and distal tubules, whereas miR-126 and miR-29b were most abundant in glomerular extracts. Subsequent experiments showed miR-126 and miR-29b enrichment in glomerular endothelial cells (GEnCs) compared with podocytes, proximal tubular epithelial cells, and fibroblasts. Significantly increased miR-126 and miR-29b were detected in GEnC conditioned medium in response to tumor necrosis factor-α and transforming growth factor-β1, respectively. Our data reveal an altered urinary miRNA profile associated with DKD and link these variations to miRNA release from GEnCs.

Elucidating genetic causes of cholestasis has proved to be important in understanding the physiology and pathophysiology of the liver. Here we show that protein-truncating mutations in the tight junction protein 2 gene (TJP2) cause failure of protein localization and disruption of tight-junction structure, leading to severe cholestatic liver disease. These findings contrast with those in the embryonic-lethal knockout mouse, highlighting differences in redundancy in junctional complexes between organs and species.

The electrical behavior of SiGe and pure Ge channel p-FET devices will be presented and discussed in this paper. By fabricating short channel SiGe, relaxed Ge and strained Ge channel devices with scaled high-κ/metal gate stacks, we demonstrate three main advantages of using these types of devices: i) more aggressive TINV scaling of the gate dielectric due to lower temperature processing, ii) band gap engineering to allow better control of threshold voltage and possibly iii) lower external resistance due to the higher B activation in these materials. The main drawback inherent to relaxed Ge and biaxially strained Ge and SiGe channels is that the ability to have improved transport properties without the need for external stressors always seems to come with a large penalty in off state leakage. The ION and IOFF in these devices are compared to recent work by others and a general trend is observed.

The aim of this study was to determine whichArabidopsis thaliana(L.) genes had significantly altered expression following 2–36 h of infestation by the aphidMyzus persicae(Sulzer). Six biological replicates were performed for both control and treatment at each time point, allowing rigorous statistical analysis of any changes. Only two genes showed altered expression after 2 h (one up- and one down-regulated) while two were down-regulated and twenty three were up-regulated at 36 h. The transcript annotation allowed classification of the significantly altered genes into a number of classes, including those involved in cell wall modification, carbon metabolism and signalling. Additionally, a number of genes were implicated in oxidative stress and defence against other pathogens. Five genes could not currently be assigned any function. The changes in gene expression are discussed in relation to current models of plant-insect interactions.

Summary 1 The consequences of the movement of transgenes from genetically modified (GM) crops into wild populations of plants continues to be of concern to ecologists and conservationists because of the possible threat posed to those populations in terms of their continued survival and because of the further knock-on effects that might occur to habitats in which they occur. 2 We examined five UK sea beet Beta vulgaris ssp. maritima populations from each of two major habitat types, cliff top and drift line. We assessed population genetic parameters, genetic diversity, gene flow, population differentiation and isolation by distance, to enable determination of the likelihood and consequences of spread to wild populations of genes from cultivated sugar beet group Beta vulgaris ssp. vulgaris, which could in the future be transgenic. 3 Drift line populations were more diverse than cliff top populations and also showed greater levels of gene flow. 4 Isolation by distance was identified in both habitats, but the relationship between genetic and geographical distance was detectable over longer distances for drift line populations. However, clear indications of vicariance (the subdivision of a population into distinct taxa by the appearance of a geological barrier) between cliff and drift line populations were also evident, because of the restriction of gene flow between the two habitats occurring more in one direction than the other. 5 Synthesis and applications. The likelihood of transgene spread from crop to wild populations is habitat dependent and conservation management decisions could therefore vary from one population to another, for example water courses were found to facilitate seed dispersal. This should be taken into account when estimating isolation distances for GM beet, and when predicting transgene frequencies (exposure estimates) for environmental risk assessments of GM beet.

The aim of this study was to identify genes that are expressed in the phloem. Increased knowledge of phloem regulation will contribute to our understanding of its many roles, from transport of solutes to information about interactions with pathogens. A cDNA library constructed from phloem-enriched sap exuding from cut Ricinus communis (L.) hypocotyls was sequenced. To assess contamination from other tissues, two libraries were constructed: one using the first 15 min of exudation and the other from sap collected after 120 min of exudation had elapsed. Of 1012 clones sequenced, 158 unique transcripts were identified. The presence of marker molecules such as profilin, the low occurrence of chloroplast-related mRNAs, and the sieve element localization of constituent mRNA using in situ hybridization were consistent with a phloem origin of the sap. Functional analysis of the cDNAs revealed classifications including ribosomal function, interaction with the environment, transport, DNA/RNA binding, and protein turnover. An analysis of the closest Arabidopsis thaliana (L.) homologue for each clone indicated that genes involved in cell localization, protein synthesis, tissue localization, organ localization, organ differentiation, and cell fate were represented at twice the level occurring in the whole Arabidopsis genome. The transcripts found in this phloem-enriched library are discussed in the context of phloem function and the relationship between the companion cell and sieve element.

We have developed an Arabidopsis thaliana/Myzus persicae model system to allow the dissection of plant/insect interactions at a molecular genetic level. This allows the examination of the role of single plant genes in the interaction between the plant and an aphid. Our initial studies have exploited an Arabidopsis genotype in which the function of the amino acid transporter ANT1 has been abolished. This mutation results in a change in the proportions of several amino acids within the phloem sieve elements (SEs) resulting in an increase in the proportion of essential amino acids. This has been measured using aphid stylectomy to collect SE samples, followed by a novel micellar electrokinetic chromatography method for amino acid analysis. The SE content represents the aphid's diet, and use of electrical penetration graph technology and honeydew clocks have demonstrated that this altered diet results in a change in the feeding rate of the aphid. Balance sheets can be produced to show the amount (nmoles/24 h) of each of 18 amino acids taken up and excreted by aphids feeding on wild type and ant1 mutant plants. The data show that aphids feeding on the ant1 mutant take up larger amounts of amino acids. However, we could not detect any effect on the reproductive rate of the aphids. The results show that, under experimental conditions, this model system can be used to identify plant genes that control the behaviour and fecundity of an insect pest.

The production of transgenic broccoli (Brassica oleracea) with increased shelf-life using an Agrobacterium rhizogenes-mediated co-transformation protocol is reported. An Agrobacterium rhizogenes Ri vector, pRi1855:GFP was constructed to allow expression of the green fluorescent protein to identify insertion of Ri TL-DNA into plant cells. The Brassica oleracea ACC synthase 1 and ACC oxidase 1 and 2 cDNAs in sense and antisense orientations were co-transformed into GDDH33, a doubled haploid calabrese-broccoli cultivar. Transformation efficiency was 3.26%, producing 150 transgenic root lines, of which 18 were regenerated into mature plants. The floral buds from T0 broccoli heads were assayed for post-harvest production of ethylene and chlorophyll levels. Buds from T0 lines transformed with ACC oxidase 1 and 2 constructs produced significantly less post-harvest ethylene at 20 °C than the untransformed plants and chlorophyll loss was significantly reduced over a 96 h post-harvest period. The T0 plants transformed with sense and antisense ACC synthase 1 had a significantly reduced 24 h post-harvest ethylene peak and delayed chlorophyll loss. A positive correlation between post-harvest bud ethylene production and chlorophyll loss was described by a regression. This demonstrates that the shelf-life of a very perishable vegetable may be increased up to 2 days at 20 °C by reducing post-harvest ethylene production.

Contact resistances are directly measured for contacts with sizes from 25 to 330 nm using e-beam based nano-TLM devices. Record low contact resistivities ~1.5 × 10-9 Ω· cm2 are extracted from Ni(Pt) silicide contacts on in situ boron-doped Si0.7Ge0.3 with a chemical boron-doping density of 2 × 1021/cm3. This is very promising for pMOS applications beyond the 10-nm node. A clear dependence of contact resistance on the silicide thickness has also been found.

Setting goals and assessing outcomes are essential elements in palliative care. This paper describes a multiprofessional project, conducted under the auspices of clinical audit, which attempted to evaluate important outcomes of care. Over a six-month period there were 123 consecutive admissions to the hospice. These patients and their carers, as well as the staff, were encouraged to set explicit goals for, and evaluate outcomes of, their care. As anticipated, we encountered many difficulties in this, but there were benefits. We were able to record goals of admission from the patient in 97 cases (79%), their main carer in 74 cases (63%) and from hospice staff in 120 cases (98%). Patient and carer goals were often more functional and specific whereas the staff goals tended to be more problem or symptom focused. The achievement of these goals was evaluated by patients, carers and staff at discharge or death (where possible) with the majority being fully or partially met. Only 15 patients and 9 carers thought that some or all of their goals had not been achieved with just 4 recording that their goals had changed. Overall, it was a worthwhile (although time-consuming) exercise and, as a result of the ‘audit’, clear goals and outcomes from patient, carer and staff perspectives are now routinely recorded for all admissions to the hospice and are used to focus multiprofessional patient review. Having analysed the process as well as the results of the ‘audit’, we would encourage others not to be daunted from undertaking similar projects.

A planar self-aligned double-gate MOSFET process has been implemented where a unique sidewall source/drain structure (S/D) permits self-aligned patterning of the back-gate layer after the S/D structure is in place. This allows coupling the silicon thickness control inherent in a planar, unpatterned layer with VLSI self-alignment techniques and also gives independently controlled front and back gates. The demanding structure led to process innovations primarily in front-end CMP, where planarity within 5 nm was achieved on an 8-in diameter wafer as well as in silicided silicon source/drain sidewalls, with minimal encroachment of the silicide. Double-gate FET (DGFET) operation is demonstrated, with good transport at both interfaces. Dense circuit layouts are achieved with multifinger devices, and logic inverters with back-gate-controlled load current as well as NOR logic using the two gates of a single transistor as inputs are demonstrated.

We have identified quantitative trait loci (QTL) for transgenic and adventitious root production using an Agrobacterium rhizogenes-mediated co-transformation system in conjunction with a Brassica oleracea double haploid (DH) mapping population. Three QTL for green fluorescent protein (GFP)-fluorescent root production and four QTL for adventitious root production were identified as accounting for 26% and 32% of the genetic variation in the population, respectively. Two of the QTL regions identified were common to both transgenic and adventitious root production. Two different methods of QTL analysis were employed (marker regression and interval mapping) and with the exception of one region on linkage group O7 for transgenic root production, both techniques detected the same regions of the genome. The regions we identified to be associated with the control of transgenic root production following A. rhizogenes-mediated transformation are the first to be detected using a QTL mapping approach. In addition, this is the first study to identify genetic regions that co-regulate both transgenic and adventitious root production within the constraints of an A. rhizogenes-mediated transformation process. We have identified plant genotypes that do not produce any transgenic roots that may be deficient for T-DNA integration via illegitimate recombination and that may also be potentially important for the development of homologous recombination protocols. Conversely, we have also identified plant genotypes with high rates of transgenic root production that will be critical in the development of high throughput transformation systems.

The production of transgenic roots was scored for eight Brassica oleracea cultivars from broccoli, cabbage, cauliflower and kale following inoculation with an Agrobacterium rhizogenes cell line carrying a binary plasmid bearing the green fluorescence protein (gfp) gene in the T-DNA. Significant differences in the numbers of explants producing transgenic roots were observed between cultivars, ranging from 1.4% for Marathon F1 to 57.8% for the Green Duke F1. Three F1 cultivars were subjected to anther culture, and doubled-haploid (DH) lines were used for transformation. The DH lines produced showed considerable variation for transgenic root production with some lines showing increased efficiency compared to the parental F1 cultivar. Grouping of the DH lines into response classes with respect to transgenic root production allowed the development of potential genetic models to explain the variation in performance released from each F1 cultivar. No apparent segregation distortion for transgenic root production was observed in the DH lines following anther culture.

Understanding the extent and distribution of genetic diversity within a species is essential for the development of effective conservation strategies. The objective of this study was to assess genetic variation using amplified fragment length polymorphisms (AFLP) in two species of the tropical legume genus Stylosanthes Sw. Annual, S. humilis (2n = 20) and perennial, S. viscosa (2n = 20) are found throughout tropical America, and are sympatric for much of their range of distribution. One hundred and eleven accessions, covering a wide geographical range, were selected for AFLP analysis. Binary data matrices derived from DNA banding patterns were analysed using the software programs NTSYS-PC and ARLEQUIN. Several accessions were found to be misidentified. Of the S. humilis accessions, the overall average similarity value was (0.72) slightly higher than the value obtained for S. viscosa (0.67). Cluster analysis and principal coordinate analysis grouped accessions from both species by geographical origin, with a few exceptions. Analysis of molecular variance (AMOVA) in S. humilis revealed 59.4% of the variation among groups formed from the cluster analysis. This was highly significant (P < 0.001). For S. viscosa AMOVA also revealed more variation among than within groups (66.5%). This was also highly significant (P< 0.001). The majority of accessions of both species conserved ex situ are of Brazilian and Venezuelan origin. This study has identified areas in Central America and Mexico for which novel genetic variation may be found and where conservation activities should be focused.

This paper addresses the potential applications of multi-protocol routing for automatic remote meter reading (ARMR). The paper concentrates on the special needs for these applications over the lower line, in terms of: the data on a lower line medium; transmitting and receiving data from any point on the low voltage distribution network; achieving reliable communications from any point on the low voltage distribution network to any other internal and external point; and the ability to increase system performance. Other communications media such as radio and telephone line are not ignored. The intention is to seek out a protocol or multiprotocol routing architecture, which could be used for power line carrier systems but equally could also he applied to these other media for the communication of metering data, and other services.

An extremely low contact resistivity of 6-7 × 10-9 Ω·cm2 between Ni0.9Pt0.1Si and heavily doped Si is achieved through Schottky barrier engineering by dopant segregation. In this scheme, the implantation of B or As is performed into silicide followed by a low-temperature drive-in anneal. Reduction of effective Schottky barrier height is manifested in the elimination of nonlinearities in IV characteristics.

Genetic diversity within populations of organisms and species is commonly measured using molecular-marker data. It has been claimed that more reliable diversity measurements can be obtained using selected genetically mapped markers to ensure that all regions of the genome are represented in the data sets employed. However, this has not been tested. In the present study, using rice (Oryza sativa L.) as a model species, we have shown that the use of unmapped AFLP markers reveals a pattern of diversity that is very similar to that obtained using a range of other marker types and which reflects the known crossability groups within this species. In contrast, we show that use of mapped-marker data can, in some cases, result in highly misleading patterns of diversity; the results obtained are critically related to the choice ofparents used in the cross from which the mapping population was produced. For diversity analyses, we propose that it is appropriate to use unmapped markers provided that the marker-type has been shown to have a wide distribution over the genome.

We have examined the effectiveness of similar numbers of markers from four molecular marker systems (AFLP, isozymes, ISSR and RAPD) for revealing genetic diversity and discriminating between infraspecific groups of Oryza sativa germplasm. Each marker system classifies the germplasm into three major groups (most effectively with isozymes and AFLPs), but with differences (primarily with ISSR) between the precise classifications generated. However, at the highest levels of genetic similarity there was only partial agreement as to relationships between individual accessions when different markers were used. When variance was partitioned among and within the three subspecific groups, although the differences were not significant, greater variation was found among than within groups using AFLP and isozymes, with the reverse for RAPD and ISSR. Measurement of polymorphism using average heterozygosity and effective number of alleles gave similar results for each marker system. These results are discussed in relation to various genetic resources conservation activities, and the advisability of extrapolating to other sets of germplasm particularly of other crop species.

An ecogeographic survey is required for the development of a strategy for the conservation and use of plant genetic resources. An herbarium-based ecogeographic survey is presented, supplemented by detailed field surveys of wildCoffea species in the Mascarene Islands. The geographical and ecological distribution of the different Coffea species in the Mascarene Islands, principally in Mauritius, is determined, and an assessment is made of the IUCN conservation status of native Coffea species. Ecogeographic data were collated from herbarium specimens from four major herbaria holding collections of Mascarene Coffea. The geography, ecology, phenology and conservation status of the Coffea species from Mauritius are discussed.

Plants were regenerated by somatic embryogenesis from long-term callus cultures derived from five garlic (Allium sativum L.) cultivars. Thirty-five of these plants were subjected to RAPD analysis. The frequency of variation was found to be cultivar dependent: approximately 1% in the two clones Solent White and California Late and around 0.35% in another three clones, Chinese, Long Keeper and Madena. Certain band changes were found in regenerants of different cultivars, suggesting the existence of a mutation-sensitive part of the garlic genome. The karyotypes of another 75 regenerants derived from the same callus cultures of three parental garlic clones were examined. Of these plants, 9.3% were found to be tetraploids, 4% aneuploid and 2.6% showed a change in the position of the secondary constriction. No association could be shown between the rate of variation for molecular and cytological characters either by comparing cultivars or examining individual regenerants.

The concept of using the power line in conjunction with the broadband integrated services network (BISDN) for a range of value added service is proposed. The use of the power line in the low voltage distribution network, as a local loop in communications for a range of customer services in the residential and industrial area is detailed. The integration of this local loop residential service into a BISDN network provides the potential for integrating many thousand of homes into a wide area network (WAN) architecture. The potential characteristics of this type of communication architecture are established for efficient transport of data services.

This paper addresses the potential applications of power-line-carrier (PLC) communications technology within the field of industrial plant/equipment control. The special needs and requirements for this application (in terms of such factors as system integrity and response times) are considered in comparison with those for other types of PLC application. Existing PLC technologies are discussed and their suitability (or otherwise) for this application considered.

The Internet was born approximately 20 years ago out of a project to integrate a US Defence Department Network called ARPAnet. This integration involved radio satellite voice and data networks. The Internet has completely revolutionised public communication networks. The concept of using the Internet for a metering communications service is proposed. The use of the Internet for a range of customer services in the residential and industrial area is also detailed. The Internet provides the potential for integrating many thousand of homes into a wide area metering communications network (WAMCN) architecture. The potential characteristics of this type of communication architecture are established for efficient transport of data services.

The sequences of ITS1 of the internal transcribed spacer regions of nuclear ribosomal DNA from 11 species or subspecies in four sections of the genus Beta were compared. Phylogeny of these wild beet taxa was inferred from the sequence data using phenetic and phylogenetic analyses. Multiple accessions from the same 11 taxa were subjected to random amplified polymorphic DNA (RAPD) analysis, and the data were analysed phenetically. With both molecular techniques and each analysis, three distinctive groups were formed: species from section Beta formed one group; species from section Procumbentes formed a very distinct group; and species from both section Nanae and section Corollinae clustered together forming the third group, which is closer to Beta than to Procumbentes. The RAPD data revealed within-section interspecies relationships that are consistent with those reported previously; this was not always the case using the single-locus sequence data.

Seven years have passed since the introduction of the liberalisation of the electricity supply industry (ESI) in the UK. Many diverse benefits and applications have been suggested which depend upon efficient bidirectional communications between the utility and its customer base. These applications have included automatic meter reading, distribution automation, load management, remote meter calibration, tokenless prepayment and remote tariff management. However the ongoing development of competition in the ESI in the UK may slow implementation, or even prevent these communications becoming a reality.

Lentil landraces from South Asia exhibit a low diversity and discordance with landraces from other countries according to a combination of qualitative and quantitative agromorphological characters. They exhibit specific phenological adaptation to the South Asian environment which precludes the direct use of alien germplasm in breeding programs in South Asia. An understanding of the genetic relationships and diversity of South Asian lentil landraces, in relation to landraces from other countries, is important in attempting to widen the genetic base of germplasm in the region. The objectives of this study were to investigate the genetic relationships between lentil landraces from 3 South Asian countries (India, Nepal and Pakistan) and those from 13 other countries and to determine their relative genetic diversities, using both isozyme electrophoresis and random amplified polymorphic DNA (RAPD) analysis. Polymorphisms were observed for 7 isozyme loci (16 alleles) and 22 RAPD loci. According to Nei's genetic distance, germplasm from Afghanistan clustered with that from the South Asian countries. The germplasm from these countries was striking different to that from the other countries studied. Based on genetic distance estimates from RAPD analysis, the countries with the lowest diversity were Pakistan, Afghanistan and Nepal. These data support evidence at the morphological level of a genetic bottleneck in lentil landraces from South Asia. Genetic relationships between countries outside the South Asian group are discussed. Classification into macrosperma and microsperma types did not reflect overall country relationships.

The key parameters to provide effective and transparent communications using the low voltage mains signalling networks are identified. The magnitude and quantitative assessment of the key parameters are discussed with respect to providing seamless communications in the utility to home and in building communications network.

No single conservation technique can adequately conserve the full range of genetic diversity of a target species or genepool. To optimize the efficiency of germplasm conservation, a range of conservation techniques needs to be applied simultaneously. In this paper we review the progress made in the application of both in situ and ex situ complementary conservation strategies and techniques to the conservation of the genus Coffea and identify areas where more research work is required. Traditionally, Coffea species have been conserved ex situ as living plants in field genebanks due to their intermediate seed storage behaviour. However, progress made in the field of biotechnology in recent years have made it possible to envisage the use of in vitro, slow growth and cryopreservation for medium- and long-term conservation of Coffea germplasm, respectively. In situ conservation is also an important option for long-term conservation, but there are no genetic reserves specifically set up for conserving Coffea. Protected areas and managed reserves offer the best opportunities for conserving Coffea. A case study for Mascarene Coffea in Mauritius is presented and a strategy for their conservation is discussed.

This paper addresses electricity supply industry deregulation in the United Kingdom and the key issues effecting the future of Regional Electricity Companies in this new environment. The development of services and communications architecture for monitoring the power distribution network in terms of power regulations are discussed. New potential services to customers are considered together with their benefits and efficiency.

This paper reviews the current standards, both from the European and international perspective, for the development of efficient methods for producing and controlling electrical energy. The formation of the different committees, their aim together with applications to control energy are discussed. The interaction of European and international bodies to provide a future unified policy for energy control is reviewed. Applications to the different potential services required in a power communication system for the future are identified.

RAPD analysis was carried out using 93 accessions held within the Oryza collection in the Genetic Resource Center at IRRI. These accessions had been designated as O. meridionalis, O. glumaepatula, O. nivara or O. rufipogon on the basis of the identification of the original collector although in some cases these had been subjected to subsequent taxonomic revision. Following numerical analysis of the RAPD data, we propose that five of the forty accessions designated as O. meridionalis and four of the 22 accessions designated as O. glumaepatula have been mis-identified. The relationship between accessions designated as O. nivara and O. rufipogon is complex, although it appears that some mis-identification has also occurred for these two taxa. The results indicate that RAPD technology can be used as a fast and accurate method to assist in the validation of the identification of wild Oryza species.

We present results from gate-all-around (GAA) silicon nanowire (SiNW) MOSFETs fabricated using a process flow capable of achieving a nanowire pitch of 30 nm and a scaled gate pitch of 60 nm. We demonstrate for the first time that GAA SiNW devices can be integrated to density targets commensurate with CMOS scaling needs of the 10 nm node and beyond. In addition, this work achieves the highest performance for GAA SiNW NFETs at a gate pitch below 100 nm.

In response to deregulation, many UK regional electricity companies (RECs) are considering redesigning their communication to meet this and other business requirements. This paper presents a proposed communication infrastructure for South Western Electricity plc. The company services a wide variety of customers in the south west of England. The supporting technology, REC and customer benefits, together with valued added services (VAS) are addressed. The SWEB communication system consists of a microwave radio network which is used for telephony, VHF radio, and data traffic. The applications of the communication system include automated meter reading, load management, quality monitoring and billing.

With an increasing competitive environment, electric utility companies face a window of both necessity and opportunity. The necessity to investigate and assess the information and telecommunications capabilities they will need to be competitive in their main operations, and the opportunity to consider new sources of revenue that such capabilities may make possible. Irrespective of approach a utility takes to integrating its main activities with the emerging information superhighway, only a confluence of energy and information technologies can meet the national challenge of efficiently managing energy demand, supply and transmission. Utilities will be driven to become more competitive in deploying both supply-side and demand side energy information services in order to control access to the customer and prevent erosion of their customer base. However at the same time, utilities have the potential to become significant players in the communications field especially towards the superhighway by providing nonenergy value added services through telecommunications infrastructures. Structuring a telecommunications architecture and strategy for a particular utility is difficult because it involves many interconnected technological, economic, competitive and financial considerations. This considers these key points including telecommunications networks, supply side information services and energy information services. The changes in architecture and technologies of communications systems leading to new services plus additional attributes to current services is covered.

The privatisation of the water, gas and electricity utilities under United Kingdom Government legislation has led to new developments of these industries in a new competitive market. Metering systems for all these utilities has developed into a very innovatory market with all three utilities investigating novel and cost effective metering communications systems. This paper discusses current field trials within the London Electricity Board in conjunction with Thorn-EMI for providing metering services to the residential customer.

Diverse Asian rice (Oryza sativa) germplasm has been used to identify associations between various quantitative traits and RAPD molecular markers using multiple regression analysis. This has allowed us to predict for other samples of germplasm their performance for traits such as culm length and number, days to flowering, grain width, and panicle and leaf length using only RAPD marker data. Such predictive capability is possible because of the availability of extensive diversity held in genebanks, and can be used in the future to facilitate the exploitation of that biodiversity. More specifically the methodology could facilitate crop improvement by rapid ideotype prediction. For the mapping and isolation of QTLs (genes controlling quantitative traits) the method would provide information to guide the selection of parental material for hybridization and markers expected to show linkage to QTLs. It may also be possible that these associations could lead the way towards marker-assisted selection during breeding programs. In the future, this demonstration of association between markers and easily measured traits could also be extended to the study of important adaptive traits, such as stress tolerance, found either within germplasm collections or in natural populations.

Annual beets in the genus Beta section Beta represent an important genetic resource. Representative accessions of annual beets from a beet germplasm collection were analysed using RAPD to assess the patterns of variation and relationships among them. Using arbitrary primers, markers showing variation across accessions were identified. A dendrogram of similarity was produced using these molecular markers. All the accessions analysed were classified into three major groups corresponding to species or subspecies macrocarpa, adanensis and maritima. Macrocarpa was shown to be the most divergent group in this section. Using RAPD molecular markers, it was possible to ascribe an accession to one of three taxonomic groups and overcome much of the confusion encountered when morphological traits are used for identification. The group of maritima was found to be more polymorphic than either the group of macrocarpa or adanensis at both accession and subspecies levels.

A set of accessions of Oryza sativa from the International Rice Research Institute (Philippines) that included known and suspected duplicates as well as closely related germplasm has been subjected to RAPD analysis. The number of primers, the number of polymorphic bands and the total number of bands were determined that will allow the accurate discrimination of these categories of accessions, including the identification of true and suspected duplicates. Two procedures have been described that could be employed on a more general basis for identifying duplicates in genetic resources collections, and further discussion on the values of such activities is presented.