bluuebunny/crossref_metadata_2025 huggingface dataset:
Direct tuber testing for Potato Y potyvirus by real‐time RT‐PCR and ELISA: reliable options for post‐harvest testing?*
Authors: Fox A. , Evans F. , Browning I.
Published: May 2005

The method currently used for testing potato tubers for viruses following harvest involves a growing‐on test. This takes up to 6 weeks to complete, and there is therefore a demand for more rapid test results. The sensitivity and reliability of direct tuber testing by DAS‐ELISA and real‐time RT‐PCR (TaqMan) were compared with the growing‐on test. In addition, the reliability of all three methods for the detection of Potato Y potyvirus (PVY) in tubers was compared over post‐harvest intervals of 6, 10, 14 and 18 weeks. The test material came from plots of tubers (cv. ‘Maris Piper’) containing a primary infection of strains PVYN and PVYO, following aphid transmission from marked infector plants grown during the 2003 season. Sample material was homogenized and divided, to provide comparative test material for detection of PVY by ELISA and real‐time RT‐PCR. Tuber eye‐plugs were then taken and subjected to the growing‐on test. The remainder of the tuber was also grown on and tested, to ensure infection was not missed as a consequence of an uneven distribution of virus throughout the tuber material. The results obtained using the two methods for direct testing of the tubers, and those results obtained from the traditional growing‐on test, are compared. The advantages and disadvantages of each method are discussed.
Comparison of a baiting method and PCR for the detection of Phytophthora fragariae var. rubi in certified raspberry stocks*
Authors: Schlenzig A. , Cooke D. E. L. , Chard J. M.
Published: May 2005

In winter 2002/2003, a total of 136 root samples from 57 different raspberry stocks in Scotland were examined for the presence of raspberry root rot caused by the fungus‐like pathogen Phytophthora fragariae var. rubi. All stocks had been planted as propagation material entered at different grades in the Scottish certification scheme or had applied for plant passports. For detection, a modified ‘Duncan bait test’ was compared to a nested PCR method. The two tests identified the same infected stocks: PCR detected 10 positive samples from four different stocks, while the bait test picked up two additional positive samples coming from the same four stocks. The two tests had a similar level of reliability in this examination and a recommendation for one or the other depends mainly on the technical equipment and skills available in the laboratory.
Identification of economically important Liriomyza species by PCR‐RFLP analysis*
Authors: Kox L. F. F. , Van Den Beld H. E. , Lindhout B. I. , De Goffau L. J. W.
Published: May 2005

Only adult males of Liriomyza bryoniae, L. huidobrensis, L. sativae and L. trifolii can be identified with certainty on basis of their genitalia. Female adults, pupae and larvae can only be identified on the level of groups of species (L. bryoniae and L. huidobrensis vs. L. sativae and L. trifolii). Species identification in all developmental stages is possible using molecular biological techniques. Our method is a PCR amplification of a 790‐bp fragment of mitochondrial cytochrome oxidase II (COII) DNA followed by RFLP analysis. The method was tested on single larvae, pupae and adults and proved to be applicable to these three life stages. The specificity of the assay was assessed by comparing the results of the PCR‐RFLP analysis with those of morphological analysis using 60 Liriomyza specimens. Molecular and morphological identification agreed for all specimens analysed. PCR‐RFLP is a powerful diagnostic tool for rapid and reliable identification of all life stages of economically important Liriomyza species.
International and regional standards for diagnosis of regulated pests*
Authors: Petter F. , Lof V. , Smith I. M.
Published: May 2005

For the last 8 years, EPPO has run a European Panel on diagnostics, which has developed regional standards on diagnostic protocols. Nearly 60 such standards have now been approved, and are in active use in EPPO countries. In 2004, the Interim Commission for Phytosanitary Measures (ICPM) of FAO, in reviewing global needs for International Standards for Phytosanitary Measures (ISPMs), recognized that there is a strong interest in developing diagnostic protocols for all IPPC contracting parties. Such protocols would support the harmonization of detection and identification procedures worldwide, contribute to greater transparency and comparability in the diagnostics of regulated pests, and assist in the resolution of disputes between trading partners. In addition, such protocols would constitute a very good material for technical assistance. In 2004, the ICPM adopted a mechanism for more rapid development of ISPMs in specific areas, suitable particularly for diagnostic protocols. A Technical Panel to develop protocols for specific pests was also established and met for the first time in September 2004. A format for international diagnostic protocols was prepared, together with a list of priority pests.
Detection of Xanthomonas fragariae in symptomless strawberry plants by nested PCR*
Authors: Moltmann E. , Zimmermann C.
Published: May 2005

Xanthomonas fragariae spreads in symptomlessly infected strawberry plantlets and a method for detection of latent infections is necessary. It is a very slow‐growing bacterium in culture and is easily overgrown by saprophytic bacteria. Therefore, plating is not a suitable method for detecting low numbers of bacteria in symptomless plants. In addition, selective media are not available. Serological assays like immunofluorescence are useful for testing in‐vitro plants, but they are not suitable for field‐grown plants, as cross reactions are common with the available antisera. For these plants, nested PCR with primers from Pooler (Pooler et al., 1996) and Zimmermann (Zimmermann et al., 2004) has proved to be a valuable method. The method was successfully applied for a survey of strawberry plants from fields in Germany and for testing imported plants (frigo and green plants).
Computer‐assisted identification and clustering for regulated phytopathogenic bacteria: construction of a reference database and development of a computer system*
Authors: Poliakoff F. , Fischer‐Le Saux M. , Hunault G. , François C. , Rivoal C. , Bonneau S. , Perret S. , Soubelet H. , Caffier D.
Published: May 2005

CABIQ is a specific computer‐assisted identification system for the reliable and rapid identification of the main regulated phytopathogenic bacteria. It is based on phenotypic and genomic properties of bacteria. About 500 reference strains have been used to initiate the database, including conventional phenotypic tests and the Biotype 100 (BioMérieux) galleries. The CABIQ system, with its database and reference matrices, is a guide on the tests to be done when identifying new isolates. The modules dealing with phenotypic and Biotype100 characteristics are already finalized. Results on repetitive PCR will soon be added to the system. This opens interesting perspectives for improving the reliability of computer‐assisted identification.
‘SSW to NNE’– North Atlantic Oscillation affects the progress of seasons across Europe
Authors: Menzel A. , Sparks T. H. , Estrella N. , Eckhardt S.
Published: May 2005

Three European plant phenological network datasets were analysed for latitudinal and longitudinal gradients of nine phenological ‘seasons’ spanning the entire year. The networks were: (1) the historical first European Phenological Network (1882–1941) by Hoffmann & Ihne, (2) the network of the International Phenological Gardens in Europe (1959–1998), founded by Schnelle & Volkert in 1957 and based on cloned plants, and (3) a dataset (1951–1998) that was recently collated during the EU Fifth Framework project POSITIVE, which included network data of seven Central and Eastern European countries. Our study is most likely the first, for over a century, to analyse average onset and year‐to‐year variability of the progress of seasons across a continent. For early, mid, and late spring seasons we found a marked progress of the seasonal onset from SW to NE throughout Europe, more precisely from WSW to ENE in early spring, then from SW to NE and finally from SSW to NNE in late spring, as exhibited by the relationship between latitudinal and longitudinal gradients. The movement of summer was more south to north directed, as the longitudinal gradient (west–east component) strongly declined or was even of opposite sign. Autumn, as shown by leaf colouring dates, arrived from NE to SW. Possible reasons for the differences among the three datasets are discussed. The annual variability of latitudinal and longitudinal gradients of the seasons across Europe was closely related to the North Atlantic Oscillation (NAO) index; in years with high NAO in both winter and spring, the west–east component of progress was more pronounced; in summer and autumn, the pattern of the seasons may be more uniform.
Tachykinins and tachykinin‐receptors in the rat pineal gland
Authors: Mukda S. , Chetsawang B. , Govitrapong P. , Schmidt P. T. , Hay‐Schmidt A. , Møller M.
Published: May 2005

High‐pressure liquid chromatography of extracts of rat pineal glands, followed by radio immunological analysis with antibodies against tachykinins, demonstrated the presence of substance P, neurokinin A and neurokinin B in the superficial rat pineal gland. Immunohistochemistry on perfusion‐fixed rat brain sections showed substance P and neurokinin A to be present in nerve fibers located both in the perivascular spaces as well as intraparenchymally between the pinealocytes. After extracting total RNA, followed by reverse transcription and polymerase chain reaction amplification with primers specific for NK1‐, NK2‐ and NK3‐receptors, agarose gel analysis of the reaction products showed the presence of mRNA encoding all three neurokinin receptors. Immunohistochemical analysis showed NK1 receptor to be located in the interstitial cells of the gland. This location was confirmed by use of in situ hybridization using radioactively labeled antisense oligonucleotide probes. Double immunohistochemical stainings showed that the NK1‐immunoreactive cells were not a part of the macrophages or antigen‐presenting cells of the gland. Our study suggests that tachykinins, after release from intrapineal nerve fibers, are involved in an up to now unknown function, different from that of melatonin synthesis.
Emotional and spatial learning in goldfish is dependent on different telencephalic pallial systems
Authors: Portavella Manuel , Vargas Juan P.
Published: May 2005

In mammals, the amygdala and the hippocampus are involved in different aspects of learning. Whereas the amygdala complex is involved in emotional learning, the hippocampus plays a critical role in spatial and contextual learning. In fish, it has been suggested that the medial and lateral region of the telencephalic pallia might be the homologous neural structure to the mammalian amygdala and hippocampus, respectively. Although there is evidence of the implication of medial and lateral pallium in several learning processes, it remains unclear whether both pallial areas are involved distinctively in different learning processes. To address this issue, we examined the effect of selective ablation of the medial and lateral pallium on both two‐way avoidance and reversal spatial learning in goldfish. The results showed that medial pallium lesions selectively impaired the two‐way avoidance task. In contrast, lateral pallium ablations impaired the spatial task without affecting the avoidance performance. These results indicate that the medial and lateral pallia in fish are functionally different and necessary for emotional and spatial learning, respectively. Present data could support the hypothesis that a sketch of these regions of the limbic system, and their associated functions, were present in the common ancestor of fish and terrestrial vertebrates 400 million years ago.
Phosphorylation of cysteine string protein in the brain: developmental, regional and synaptic specificity
Authors: Evans Gareth J. O. , Morgan Alan
Published: May 2005

Protein phosphorylation modulates regulated exocytosis in most cells, including neurons. Cysteine string protein (CSP) has been implicated in this process because its phosphorylation on Ser10 alters its interactions with syntaxin and synaptotagmin, and because the effect of CSP overexpression on exocytosis kinetics in chromaffin cells requires phosphorylatable Ser10. To characterize CSP phosphorylation in the brain, we raised phosphospecific antibodies to Ser10. Western blotting revealed that the proportion of phosphorylated CSP (P‐CSP) varies between distinct brain regions and also exhibits developmental regulation, with P‐CSP highest early in development. Immunohistochemical analysis of the cerebellar cortex revealed a novel pool of P‐CSP that did not colocalize with synaptic vesicle markers during early development. Strikingly, in the adult cerebellar granular layer P‐CSP was highly enriched in a subset of glutamatergic synapses but undetectable in neighbouring GABA‐ergic synapses. In view of the functional consequences of CSP phosphorylation, such differences could contribute to the synapse‐specific regulation of neurotransmitter release.
Hepatic vagotomy alters limbic and hypothalamic neuropeptide responses to insulin‐dependent diabetes and voluntary lard ingestion
Authors: La Fleur Susanne E. , Manalo Sotara L. , Roy Monica , Houshyar Hani , Dallman Mary F.
Published: May 2005

Hypothalamic anorexigenic [corticotropin‐releasing factor (CRF) and proopiomelanocortin] peptides decrease and the orexigen, neuropeptide Y, increases with diabetic hyperphagia. However, when diabetic rats are allowed to eat lard (saturated fat) as well as chow, both caloric intake and hypothalamic peptides normalize. These neuropeptide responses to lard require an intact hepatic vagus [la Fleur et al. (2003)Diabetes, 52, 2321–2330]. Here, we delineate temporal interactions after lard consumption ± hepatic vagotomy (HV) between feeding and brain neuropeptide expression in insulin‐dependent diabetic rats. CRF‐mRNA was reduced in the paraventricular nuclei (PVN) by 6 h after presentation of lard, before caloric intake increased in HV‐diabetic rats, and did not increase at 30 or 36 h, as it did in shamHV‐diabetic rats eating lard. CRF‐mRNA was increased in the bed nuclei of the stria terminalis of HV‐diabetic rats compared with shamHV‐diabetic rats only when caloric intake was high at 30 or 36 h. At 36 h, shamHV‐diabetic rats eating chow had increased CRF‐mRNA in the central amygdala but diabetic rats eating lard had decreased CRF‐mRNA, whereas HV‐diabetic rats eating chow or lard had normal CRF‐mRNA in the central amygdala. We conclude that eating lard restores peptide expression to normal in the hypothalamus of diabetic rats, and because decreased CRF‐mRNA in the PVN precedes the increase in caloric intake in HV‐diabetic rats eating lard, that the loss of a hepatic vagal signal to PVN may be responsible for increased intake; moreover, CRF‐mRNA in limbic structures is also sensitive to both HV and lard ingestion in diabetic rats.
Congenital anomaly rate in offspring of mothers with diabetes treated with insulin lispro during pregnancy
Authors: Wyatt J. W. , Frias J. L. , Hoyme H. E. , Jovanovic L. , Kaaja R. , Brown F. , Garg S. , Lee‐Parritz A. , Seely E. W. , Kerr L. , Mattoo V. , Tan M. ,   the IONS study group
Published: May 2005

Aim  To determine the rate of major congenital anomalies in offspring of a large group of women with diabetes mellitus treated with insulin lispro (Humalog®).

Methods  This multinational, multicentre, retrospective study included mothers with diabetes mellitus (diagnosed prior to conception) who were treated with insulin lispro for at least 1 month before conception and during at least the first trimester of pregnancy. Anomalies were assessed by two independent dysmorphologists not affiliated with the sponsor.

Results  The charts of 496 women were reviewed for 533 pregnancies resulting in 542 offspring (500 live births, 31 spontaneous and seven elective abortions, and four stillbirths). Mothers’ characteristics: mean (± sd) age was 29.9 (± 5.2) years, 85.6% were Caucasian and 97.2% had Type 1 diabetes mellitus. Insulin lispro continued to be the main mealtime insulin for more than 96% of the women during the second and third trimester. The dysmorphologists determined that 27 (5.4%) offspring had major congenital anomalies and 2 (0.4%) offspring had minor congenital anomalies.

Conclusions  The rate of major congenital anomalies was 5.4%[95% CI (3.45%, 7.44%)] for offspring of mothers with diabetes mellitus treated with insulin lispro before and during pregnancy. The current published rates of major anomalies in infants born to mothers with diabetes treated with insulin are between 2.1 and 10.9%. This suggests that the anomaly rate with insulin lispro treatment does not differ from the published major congenital anomaly rates for other insulin treatments.
In situ U–Pb dating of zircon formed from retrograde garnet breakdown during decompression in Rogaland, SW Norway
Authors: TOMKINS H. S. , WILLIAMS I. S. , ELLIS D. J.
Published: May 2005

Regionally metamorphosed metapelites from Rogaland, SW Norway, contain zircon formed during the decompression reaction garnet + sillimanite + quartz → cordierite. The zircon, which occurs as inclusions in cordierite coronas around garnet, is texturally, chemically and isotopically distinct from older zircon in other textural settings in the matrix. A SHRIMP U–Pb age of 955 ± 8 Ma based on analyses in thin section on the decompression zircon from the cordierite coronas, therefore dates a point on the retrograde path, estimated from garnet–cordierite equilibria to be 5.6 kbar, 710 °C. This population was under‐represented in conventional SHRIMP analyses of individual zircon in a mono‐mineralic grain mount and, in the absence of a textural context, its significance unknown. The dominant age identified from SHRIMP analyses of the grain mount, in combination with analyses from matrix zircon in thin section, was 1035 ± 9 Ma. Based on the lack of consistent textural relationships with any specific minerals in thin section, as well as rare earth element chemistry, the 1035‐Ma population is interpreted to represent zircon growth during incipient migmatization of the rocks at 6–8 kbar and c. 700 °C. This is consistent with previous estimates for the age of regional M1 metamorphism during the Sveconorwegian Orogeny. The most important outcome of this study is the successful analysis of zircon grains in a specific, well‐constrained reaction texture. Not only does this allow a precise point on the regional P–T path to be dated, but it also emphasizes the possibility of zircon formation during the retrograde component of a typical metamorphic cycle.
Oxide and sulphide isograds along a Late Archean, deep‐crustal profile in Tamil Nadu, south India
Authors: HARLOV D. E. , HANSEN E. C.
Published: May 2005

Oxide–sulphide–Fe–Mg–silicate and titanite–ilmenite textures as well as their mineral compositions have been studied in felsic and intermediate orthogneisses across an amphibolite (north) to granulite facies (south) traverse of lower Archean crust, Tamil Nadu, south India. Titanite is limited to the amphibolite facies terrane where it rims ilmenite or occurs as independent grains. Pyrite is widespread throughout the traverse increasing in abundance with increasing metamorphic grade. Pyrrhotite is confined to the high‐grade granulites. Ilmenite is widespread throughout the traverse increasing in abundance with increasing metamorphic grade and occurring primarily as hemo‐ilmenite in the high‐grade granulite facies rocks. Magnetite is widespread throughout the traverse and is commonly associated with ilmenite. It decreases in abundance with increasing metamorphic grade. In the granulite facies zone, reaction rims of magnetite + quartz occur along Fe–Mg silicate grain boundaries. Magnetite also commonly rims or is associated with pyrite. Both types of reaction rims represent an oxidation effect resulting from the partial subsolidus reduction of the hematite component in ilmenite to magnetite. This is confirmed by the presence of composite three oxide grains consisting of hematite, magnetite and ilmenite. Magnetite and magnetite–pyrite micro‐veins along silicate grain boundaries formed over a wide range of post‐peak metamorphic temperatures and pressures ranging from high‐grade SO2 to low‐grade H2S‐dominated conditions. Oxygen fugacities estimated from the orthopyroxene–magnetite–quartz, orthopyroxene–hematite–quartz, and magnetite–hematite buffers average 2.5 log units above QFM. It is proposed that the trends in mineral assemblages, textures and composition are the result of an external, infiltrating concentrated brine containing an oxidizing component such as CaSO4 during high‐grade metamorphism later acted upon by prograde and retrograde mineral reactions that do not involve an externally derived fluid phase.
Flow kinematics in the deeper part of a subduction channel, as inferred from inclusion trails in plagioclase porphyroblasts from the Sambagawa metamorphic rocks, south‐west Japan
Authors: TAKESHITA T. , ISHII K. , KANAGAWA K.
Published: May 2005

The subduction and exhumation of accretionary prism metasedimentary rocks are accompanied by large‐strain ductile deformations which may be recorded in microstructures. Porphyroblast microstructures have been a key to unravel the kinematics in such deformed belts. Shape‐preferred orientation (SPO) of epidote and amphibole inclusions that define S‐shaped trails in prograde cores of plagioclase porphyroblasts were analysed from the high‐P/T Sambagawa metamorphic rocks. Inclusions are found to be elongate parallel to the [010] and [001] directions, respectively, and their long‐axis orientations define an internal foliation Si (best‐fit great circle) and lineation Li (maximum on the Si). S‐shaped inclusion trails in the orthogonal sections do not exhibit the same geometries, but rather are grouped into two types, where the foliation intersection axes (FIAs) are nearly perpendicular and parallel to Li, respectively. These two types of S‐shaped inclusion trails are seen in the sections inclined at low and high angles to the Li, respectively. However, the latter type commonly consists of composite trails, where the Si is first rotated about an FIA perpendicular to the Li (i.e. unique axis), and then about an FIA parallel to the Li. The S‐shaped inclusion trails are interpreted to have formed by the successive overgrowth of matrix minerals and rotation of the plagioclase porphyroblast cores about a unique axis in non‐coaxial deformation. The rotation of Si about an FIA nearly parallel to the Li is perhaps an apparent rotation, caused by the deflection of foliation around the growing prismatic plagioclase grain prior to inclusion into the porphyroblast. This study has for the first time documented the 3‐D geometry of S‐shaped inclusion trails in porphyroblasts from accretionary prism metasedimentary rocks and identified their origin, which helps to understand the flow kinematics in the deeper part of a subduction channel.
Behavior and the limits of genomic plasticity: power and replicability in microarray analysis of honeybee brains
Authors: Cash A. C. , Whitfield C. W. , Ismail N. , Robinson G. E.
Published: May 2005

Transcription is slow relative to many post‐transcriptional processes in the brain. Using the rich system of division of labor in the honeybee (Apis mellifera), we found extreme differences in the extent to which behavioral occupations of different durations were associated with gene‐expression differences in the brain. Nursing and foraging, occupations lasting >1 week, were associated with significant expression differences for nearly one‐quarter of the genes tested (1208 of 5563 cDNAs tested; P < 0.01, anova), consistent with previous results. In contrast, transitional occupations, performed for 1–2 days after nursing and before the onset of foraging, were associated with either no differences (guards vs. undertakers; 19 cDNAs, fewer than the expectation of 56 false‐positives) or few differences (comb builders vs. guards and undertakers; 248 cDNAs), but extensive differences relative to both nursing and foraging (>500 cDNAs, all contrasts). Statistical power analysis indicated that expression differences of two‐, 1.5‐ and 1.25‐fold should have been detected in 100, 92 and 37% of cases, respectively. Replication of previous results at these magnitudes was 95, 71 and 51%, with no genes showing differences in the opposite direction. These results indicate that behavioral plasticity over different time‐scales may be associated with substantial differences in the extent of genomic plasticity in the brain.
Triggering Parent Company Liability Under United States Sanctions Regimes: The Troubling Implications of Prohibiting Approval and Facilitation
Authors: Lau Terence J.
Published: May 2005

[General Electric] conducts business in more than 100 countries around the world… . An important challenge for all of us is to understand how [laws of different countries] apply to our operations. GE, the parent company, is a corporation organized in the United States. The laws of the United States frequently extend to the operations of GE and its affiliates throughout the world as well as to the business activities of GE employees wherever they live and work. Other countries may also apply their own laws outside of their borders to their own citizens and to corporations that are organized under their laws, such as GE subsidiaries or other controlled affiliates… . In some instances, there may be a conflict between the applicable laws of two or more countries. When you encounter such a conflict, it is especially important to consult company legal counsel to understand how to resolve that conflict properly.1
Use of tracers and isotopes to evaluate vulnerability of water in domestic wells to septic waste
Authors: Verstraeten I.M. , Fetterman G.S. , Meyer M.T. , Bullen T. , Sebree S.K.
Published: May 2005

In Nebraska, a large number (>200) of shallow sand‐point and cased wells completed in coarse alluvial sediments along rivers and lakes still are used to obtain drinking water for human consumption, even though construction of sand‐point wells for consumptive uses has been banned since 1987. The quality of water from shallow domestic wells potentially vulnerable to seepage from septic systems was evaluated by analyzing for the presence of tracers and multiple isotopes. Samples were collected from 26 sand‐point and perforated, cased domestic wells and were analyzed for bacteria, coliphages, nitrogen species, nitrogen and boron isotopes, dissolved organic carbon (DOC), prescription and nonprescription drugs, or organic waste water contaminants. At least 13 of the 26 domestic well samples showed some evidence of septic system effects based on the results of several tracers including DOC, coliphages, NH4+, NO3−, N2, δ15N[NO3−] and boron isotopes, and antibiotics and other drugs. Sand‐point wells within 30 m of a septic system and <14 m deep in a shallow, thin aquifer had the most tracers detected and the highest values, indicating the greatest vulnerability to contamination from septic waste.
Trichlorofluoroethene: A reactive tracer for evaluating reductive dechlorination in large‐diameter permeable columns
Authors: Field Jennifer A. , Reed Ralph L. , Istok Jonathan D. , Semprini Lewis , Bennett Peter , Buscheck Timothy E.
Published: May 2005

Trichlorofluoroethene (TCFE) was used as a reactive tracer to determine the in situ rate of reductive dechlorination in treatment zones impacted by three large‐diameter permeable columns (LDPCs) that were installed at a trichloroethene (TCE)–contaminated site. The LDPCs were part of a pilot study to evaluate the effectiveness of hydrogen, lactate, and zero‐valent iron for remediating TCE‐contaminated ground water. The rate of TCFE reductive dechlorination was determined for each LDPC by means of push‐pull tests conducted in each treatment layer. In addition, the distribution of TCFE's lesser chlorinated transformation products was determined. The rates of TCFE reductive dechlorination ranged from 0.05/d to 0.20/d and corresponded to half‐lives ranging from 3.5 to 13.9 d. cis‐Dichlorofluoroethene was the dominant transformation product detected in all the tests, which is consistent with the findings from pilot tests conducted in the LDPCs prior to the TCFE push‐pull tests. cis‐Chlorofluoroethene (CFE) and 1,1‐CFE also were detected and indicate the potential for vinyl chloride to form under all treatment regimes. Significant production of fluoroethene (FE), the analog of ethene, was observed for only one of the hydrogen treatments. Unambiguous and sensitive detection of the lesser chlorinated products, such as CFE and FE, is possible because TCFE and its transformation products are not found in the background ground water at contaminated sites. Good agreement between the rates and transformation product profiles for TCFE and TCE in both field and laboratory experiments indicates the suitability of TCFE as a surrogate for predicting the rates of TCE reductive dechlorination.
Real‐time model parameter estimation for analyzing transport in porous media
Authors: Kim Juyoul , Park Yeonjeong , Harmon Thomas C.
Published: May 2005

This work describes the integration of data acquisition (DAQ) hardware and software for the purpose of acquiring not only data but real‐time transport model parameter estimates in the context of subsurface flow and transport problems. Integrated DAQ parameter estimation systems can be used to reduce data storage requirements, trigger event recognition and more detailed sampling actions, and otherwise enhance remote monitoring capabilities. The contaminant transport problem is posed here as the analogous heat transfer problem in a three‐dimensional, intermediate‐scale physical aquifer model. A constant source of warm water is fed into a sandy aquifer undergoing steady, unidirectional flow. The spatial distribution of temperature in the medium is monitored over time using 17 thermocouples embedded in the medium. These sensors log temperatures via conventional analog‐to‐digital conversion hardware driven by commercially available DAQ software (LabVIEW™). Parameter estimation routines programmed in MATLAB™‐based M‐files are embedded in the LabVIEW DAQ routine and access parameter estimation libraries, such as the descent method employed here, via the Internet. The integrated DAQ parameter estimation system is demonstrated for the estimation of (1) the thermal dispersion coefficients (analogous to mass dispersion coefficients), given a known heat source; and (2) the location of a heat source, given known thermal dispersion coefficients. In both cases, the parameter estimation procedure is executed repeatedly as the data are acquired. For the case of source location, the effect of the number of sensors on the parameter estimation procedure is also demonstrated. Reasonable parameter estimates are provided rapidly during both the transient and steady‐state phases of the experiments, with accuracy increasing with time and with the number of observations employed.
Cost‐optimal contaminant plume management with a combination of pump‐and‐treat and physical barrier systems
Authors: Bayer Peter , Finkel Michael , Teutsch Georg
Published: May 2005

The economic potential of combining hydraulic barriers and pump‐and‐treat systems to manage contaminant plumes is investigated by means of a comparative cost analysis. The analysis is based on a preceding study on the hydraulic performance of barrier‐supported and conventional pump‐and‐treat systems, which revealed the efficiency of accurately positioned barriers in reducing pumping rates for contaminant plume capture. In the present paper, we examine whether the application of physical barriers like slurry walls or sheet piles also leads to economic benefits, i.e., whether the reduction of the operational costs outweighs the capital costs associated with the construction of the barriers. An economic model is presented that quantifies remediation costs by the use of cost functions relating costs to site‐specific and system‐specific parameters. The model is applied to a hypothetical remediation scenario. The results clearly demonstrate that physical barriers may yield significant savings in the total costs, particularly if unit costs for on‐site treatment are high. Furthermore, as barriers tend to reduce the uncertainty about the pumping rate required to capture a plume, they smooth the trade‐off curves between total remediation costs and system reliability.
Phenformin and 5‐aminoimidazole‐4‐carboxamide‐1‐β‐D‐ribofuranoside (AICAR) activation of AMP‐activated protein kinase inhibits transepithelial Na+ transport across H441 lung cells
Authors: Woollhead Alison M. , Scott John W. , Hardie D. Grahame , Baines Deborah L.
Published: May 2005

Active re‐absorption of Na+ across the alveolar epithelium is essential to maintain lung fluid balance. Na+ entry at the luminal membrane is predominantly via the amiloride‐sensitive Na+ channel (ENaC) down its electrochemical gradient. This gradient is generated and maintained by basolateral Na+ extrusion via Na+,K+‐ATPase an energy‐dependent process. Several kinases and factors that activate them are known to regulate these processes; however, the role of AMP‐activated protein kinase (AMPK) in the lung is unknown. AMPK is an ultra‐sensitive cellular energy sensor that monitors energy consumption and down‐regulates ATP‐consuming processes when activated. The biguanide phenformin has been shown to independently decrease ion transport processes, influence cellular metabolism and activate AMPK. The AMP mimetic drug 5‐aminoimidazole‐4‐carboxamide‐1‐β‐d‐ribofuranoside (AICAR) also activates AMPK in intact cells. Western blotting revealed that both the α1 and α2 catalytic subunits of AMPK are present in Na+ transporting H441 human lung epithelial cells. Phenformin and AICAR increased AMPK activity in H441 cells in a dose‐dependent fashion, stimulating the kinase maximally at 5–10 mm (P= 0.001, n= 3) and 2 mm (P < 0.005, n= 3), respectively. Both agents significantly decreased basal ion transport (measured as short circuit current) across H441 monolayers by approximately 50% compared with that of controls (P < 0.05, n= 4). Neither treatment altered the resistance of the monolayers. Phenformin and AICAR significantly reduced amiloride‐sensitive transepithelial Na+ transport compared with controls (P < 0.05, n= 4). This was a result of both decreased Na+,K+‐ATPase activity and amiloride‐sensitive apical Na+ conductance. Transepithelial Na+ transport decreased with increasing concentrations of phenformin (0.1–10 mm) and showed a significant correlation with AMPK activity. Taken together, these results show that phenformin and AICAR suppress amiloride‐sensitive Na+ transport across H441 cells via a pathway that includes activation of AMPK and inhibition of both apical Na+ entry through ENaC and basolateral Na+ extrusion via the Na+,K+‐ATPase. These are the first studies to provide a cellular signalling mechanism for the action of phenformin on ion transport processes, and also the first studies showing AMPK as a regulator of Na+ absorption in the lung.
Visualization of transmitter release with zinc fluorescence detection at the mouse hippocampal mossy fibre synapse
Authors: Qian Jing , Noebels Jeffrey L.
Published: May 2005

Exocytosis of synaptic vesicle contents defines the quantal nature of neurotransmitter release. Here we developed a technique to directly assess exocytosis by measuring vesicular zinc release with the zinc‐sensitive dye FluoZin‐3 at the hippocampal mossy fibre (MF) synapse. Using a photodiode, we were able to clearly resolve the zinc fluorescence transient ([Zn2+]t) with a train of five action potentials in mouse hippocampal brain slices. The vesicular origin of [Zn2+]t was verified by the lack of zinc signal in vesicular zinc transporter Znt3‐deficient mice. Manipulating release probability with the application of neuromodulators such as DCG IV, 4‐aminopyridine and forskolin as well as a paired train stimulation protocol altered both the [Zn2+]t and the field excitatory postsynaptic potential (fEPSP) coordinately, strongly indicating that zinc is co‐released with glutamate during exocytosis. Since zinc ions colocalize with glutamate in small clear vesicles and modulate postsynaptic excitability at NMDA and GABA receptors, the findings establish zinc as a cotransmitter during physiological signalling at the mossy fibre synapse. The ability to directly visualize release dynamics with zinc imaging will facilitate the exploration of the molecular pharmacology and plasticity of exocytosis at MF synapses.
Inverted photocurrent responses from amphibian rod photoreceptors: role of membrane voltage in response recovery
Authors: Jarvinen Jaakko L. P. , Lamb T. D.
Published: May 2005

We recorded photocurrent responses of retinal rods isolated from cane toads Bufo marinus and clawed frogs Xenopus laevis. With the outer segment drawn part way into the suction pipette, presentation of flashes to the base of the outer segment (outside the pipette) elicited a slow inverted response. Stimulation of the same region, with the outer segment drawn fully in, gave a response of conventional polarity. For moderate to bright flashes a fast transient preceded the slow inverted response. Upon bleaching the tip of the outer segment, the slow inverted response was abolished but the fast initial transient remained, and we attribute this fast component to a capacitive current. Experiments employing simultaneous whole‐cell patch‐clamp and suction pipette recording revealed that both the fast and slow components of the inverted responses were absent in voltage‐clamped cells. In current‐clamped cells the slow inverted current response was delayed substantially with respect to the voltage response. We present a computational model for the slow component, in which hyperpolarization leads to increased activity of the Na+–Ca2+,K+ exchanger, hence lowering the cytoplasmic Ca2+ concentration, activating guanylyl cyclase, raising cyclic GMP concentration, opening cyclic nucleotide‐gated channels, and increasing circulating current in the unstimulated region. For the measured voltage response to stimulation of the base, we solve these equations to predict the photocurrent in the tip, and obtain an adequate explanation of the inverted responses. Our work suggests a novel role for membrane voltage in accelerating the inactivation phase of the response to light.
Role of phospholipase D and diacylglycerol in activating constitutive TRPC‐like cation channels in rabbit ear artery myocytes
Authors: Albert A. P. , Piper A. S. , Large W. A.
Published: May 2005

Previously we have described a constitutively active Ca2+‐permeable non‐selective cation channel in freshly dispersed rabbit ear artery myocytes that has similar properties to canonical transient receptor potential (TRPC) channel proteins. In the present study we have investigated the transduction pathways responsible for stimulating constitutive channel activity in these myocytes. Application of the pharmacological inhibitors of phosphatidylcholine‐phospholipase D (PC‐PLD), butan‐1‐ol and C2 ceramide, produced marked inhibition of constitutive channel activity in cell‐attached patches and also butan‐1‐ol produced pronounced suppression of resting membrane conductance measured with whole‐cell recording whereas the inactive isomer butan‐2‐ol had no effect on constitutive whole‐cell or channel activity. In addition butan‐1‐ol had no effect on channel activity evoked by the diacylglycerol (DAG) analogue 1‐oleoyl‐2‐acetyl‐sn‐glycerol (OAG). Inhibitors of PC‐phospholipase C (PC‐PLC) and phospholipase A2 (PLA2) had no effect on constitutive channel activity. Application of a purified PC‐PLD enzyme and its metabolite phosphatidic acid to inside‐out patches markedly increased channel activity. The phosphatidic acid phosphohydrolase (PAP) inhibitor dl‐propranolol also inhibited constitutive and phosphatidic acid‐induced increases in channel activity but had no effect on OAG‐evoked responses. The DAG lipase and DAG kinase inhibitors, RHC80267 and R59949 respectively, which inhibit DAG metabolism, produced transient increases in channel activity which were mimicked by relatively high concentrations (40 μm) of OAG. The protein kinase C (PKC) inhibitor chelerythrine did not prevent channel activation by OAG but blocked the secondary inhibitory response of OAG. It is proposed that endogenous DAG is involved in the activation of channel activity and that its effects on channel activity are concentration‐dependent with higher concentrations of DAG also inhibiting channel activity through activation of PKC. This study indicates that constitutive cation channel activity in ear artery myocytes is mediated by DAG which is generated by PC‐PLD via phosphatidic acid which represents a novel activation pathway of cation channels in vascular myocytes.
Is conceiving a child to benefit another against the interests of the new child?
Authors: Spriggs M
Published: May 2005

Conceiving a child by way of embryo selection and tissue matching to benefit a sick sibling is generally justified on the grounds that as well as the potential to save the sick child, there is a benefit for the new baby. The new baby is selected so he or she will not have the disease suffered by the first child. It is not possible, however, to select against conditions for which there is no test and Jamie Whitaker’s birth is a case where the process of in vitro fertilisation with tissue matching is viewed as being of benefit only to a third party—the sick child. Some people object to using the technology for this purpose. There are also good reasons to argue that the technology should be used to save a sick child, and that it would be morally remiss for Jamie’s parents not to consent to the use of his cord blood.
HIV exceptionalism, CD4+ cell testing, and conscientious subversion
Authors: Jansen L A
Published: May 2005

In recent years, many states in the United States have passed legislation requiring laboratories to report the names of patients with low CD4 cell counts to their state Departments of Health. This name reporting is an integral part of the growing number of “HIV Reporting and Partner Notification Laws” which have emerged in response to recently revised guidelines suggested by the National Centers for Disease Control (CDC). Name reporting for patients with low CD4 cell counts allows for a more accurate tracking of the natural history of HIV disease. However, given that this test is now considered to be an “indicator” of HIV, should it be subject to the same strict consent required for HIV testing? While the CDC has recommended that each state develop its own consent requirements for CD4 cell testing, most states have continued to rely on the presumed consent standards for CD4 cell testing that were in place before the passage of name reporting statutes. This allows physicians who treat patients who refuse HIV testing to order a CD4 cell blood analysis to gather information that is indicative of their patient’s HIV status. This paper examines the ethical and legal issues associated with the practice of “conscientious subversion” as it arises when clinicians use CD4 cell counts as a surrogate for HIV testing.
The current state of clinical ethics and healthcare ethics committees in Belgium: Table 1
Authors: Meulenbergs T , Vermylen J , Schotsmans P T
Published: May 2005

Ethics committees are the most important practical instrument of clinical ethics in Belgium and fulfil three tasks: the ethical review of experimental protocols, advising on the ethical aspects of healthcare practice, and ethics consultation. In this article the authors examine the current situation of ethics committees in Belgium from the perspective of clinical ethics. Firstly, the most important steps which thus far have been taken in Belgium are examined. Secondly, recent opinion by the Belgian Advisory Committee on Bioethics with regard to ethics committees is presented and the activities of Belgian ethics committees are discussed. Finally, the option to bring research ethics and clinical ethics under the roof of just one committee is criticised using a pragmatic and a methodological argument. Concomitantly, the authors build an argument in favour of the further development of ethics consultation.
How medical ethical principles are applied in treatment with artificial insemination by donors (AID) in Hunan, China: effective practice at the Reproductive and Genetic Hospital of CITIC-Xiangya
Authors: Li L J , Lu G X
Published: May 2005

This paper investigates the efficiency of application of medical ethics principles in the practice of artificial insemination by donors (AID) in China, in a culture characterised by traditional ethical values and disapproval of AID. The paper presents the ethical approach to AID treatment as established by the Reproduction and Genetics Hospital of CITIC-Xiangya (CITIC Hunan-Yale Approach) in the central southern area of China against the social ethical background of China and describes its general features. The CITIC-Xiangya Approach facilitates the implementation of ethical relations between clinicians and patients participating in AID treatment procedures in Hunan-Yale.
The acceptability of ending a patient’s life
Authors: Guedj M , Gibert M , Maudet A , Muñoz Sastre M T , Mullet E , Sorum P C
Published: May 2005

Objectives:To clarify how lay people and health professionals judge the acceptability of ending the life of a terminally ill patient.

Design:Participants judged this acceptability in a set of 16 scenarios that combined four factors: the identity of the actor (patient or physician), the patient’s statement or not of a desire to have his life ended, the nature of the action as relatively active (injecting a toxin) or passive (disconnecting life support), and the type of suffering (intractable physical pain, complete dependence, or severe psychiatric illness).

Participants:115 lay people and 72 health professionals (22 nurse’s aides, 44 nurses, six physicians) in Toulouse, France.

Main measurements:Mean acceptability ratings for each scenario for each group.

Results:Life ending interventions are more acceptable to lay people than to the health professionals. For both, acceptability is highest for intractable physical suffering; is higher when patients end their own lives than when physicians do so; and, when physicians are the actors, is higher when patients have expressed a desire to die (voluntary euthanasia) than when they have not (involuntary euthanasia). In contrast, when patients perform the action, acceptability for the lay people and nurse’s aides does not depend on whether the patient has expressed a desire to die, while for the nurses and physicians unassisted suicide is more acceptable than physician assisted suicide.

Conclusions:Lay participants judge the acceptability of life ending actions in largely the same way as do healthcare professionals.
A model for scoring and grading willingness of a potential living related donor: Table 1
Authors: Al-Khader A A
Published: May 2005

There are few examples in the literature of objective measures for the assessment of donor willingness. The author describes the scoring system in use at his own renal transplant unit which has brought objectivity to the process of determining the willingness of living related donors. In this system, a total score to determine the degree of willingness or unwillingness is calculated based on responses to a series of questions. The author believes that with minor modifications this system could be implemented by transplant units in different countries and cultures to screen out donors who are acting under duress.