bluuebunny/crossref_metadata_2025 huggingface dataset:
Severe HDL deficiency due to novel defects in the ABCA1 transporter
Authors: Pisciotta L. , Bocchi L. , Candini C. , Sallo R. , Zanotti I. , Fasano T. , Chakrapani A. , Bates T. , Bonardi R. , Cantafora A. , Ball S. , Watts G. , Bernini F. , Calandra S. , Bertolini S.
Published: February 2009

Objectives.  The objective was the identification and functional characterization of mutations in the ABCA1 gene in four patients with severe HDL deficiency.

Subjects.  Patients were referred to the clinic because of almost complete HDL deficiency.

Methods.  The ABCA1 gene was sequenced directly. The analysis of the ABCA1 protein, ABCA1 mRNA and ABCA1‐mediated cholesterol efflux was performed in cultured fibroblasts. Intracellular localization of ABCA1 mutants was investigated in transfected HEK293 cells.

Results.  Two patients were homozygous for mutations in the coding region of the ABCA1 gene, resulting in an amino acid substitution (p.A1046D) and a truncated protein (p.I74YFsX76). The third patient was homozygous for a splice site mutation in intron 35 (c.4773 + 1g>a), resulting in an in‐frame deletion of 25 amino acids (del p.D1567_K1591) in ABCA1. These patients had clinical manifestations of accumulation of cholesterol in the reticulo‐endothelial system. The fourth patient, with preclinical atherosclerosis, was a compound heterozygote for two missense mutations (p.R587W/p.W1699C). ABCA1‐mediated cholesterol efflux was abolished in fibroblasts from patients with p.A1046D and del p.D1567_K1591 mutants and in fibroblasts homozygous for p.R587W. A reduced ABCA1 protein content was observed in these cells, suggesting an increased intracellular degradation. The mutant p.W1699C was largely retained in the endoplasmic reticulum, when expressed in HEK293 cells.

Conclusions.  The homozygotes for mutations which abolish ABCA1 function showed overt signs of involvement of the reticulo‐endothelial system. This was not the case in the compound heterozygote for missense mutations, suggesting that this patient retains some residual ABCA1 function that reduces cholesterol accumulation in the reticulo‐endothelial system.
High‐frequency field measurements of diurnal carbon isotope discrimination and internal conductance in a semi‐arid species, Juniperus monosperma
Authors: BICKFORD CHRISTOPHER P. , MCDOWELL NATE G. , ERHARDT ERIK B. , HANSON DAVID T.
Published: February 2009

We present field observations of carbon isotope discrimination (Δ) and internal conductance of CO2 (gi) collected using tunable diode laser spectroscopy (TDL). Δ ranged from 12.0 to 27.4‰ over diurnal periods with daily means from 16.3 ± 0.2‰ during drought to 19.0 ± 0.5‰ during monsoon conditions. We observed a large range in gi, with most estimates between 0.04 and 4.0 µmol m−2 s−1 Pa−1. We tested the comprehensive Farquhar, O'Leary and Berry model of Δ (Δcomp), a simplified form of Δcomp (Δsimple) and a recently suggested amendment (Δrevised). Sensitivity analyses demonstrated that varying gi had a substantial effect on Δcomp, resulting in mean differences between observed Δ (Δobs) and Δcomp ranging from 0.04 to 9.6‰. First‐order regressions adequately described the relationship between Δ and the ratio of substomatal to atmospheric CO2 partial pressure (pi/pa) on all 3 d, but second‐order models better described the relationship in July and August. The three tested models each best predicted Δobs on different days. In June, Δsimple outperformed Δcomp and Δrevised, but incorporating gi and all non‐photosynthetic fractionations improved model predictions in July and August.
Carbon‐isotope records of the Early Jurassic (Toarcian) oceanic anoxic event from the Valdorbia (Umbria–Marche Apennines) and Monte Mangart (Julian Alps) sections: palaeoceanographic and stratigraphic implications
Authors: SABATINO NADIA , NERI RODOLFO , BELLANCA ADRIANA , JENKYNS HUGH C. , BAUDIN FRANÇOIS , PARISI GUIDO , MASETTI DANIELE
Published: February 2009

The Toarcian oceanic anoxic event (ca 183 Ma) coincides with a global perturbation marked by enhanced organic carbon burial and a general decrease in calcium carbonate production, probably triggered by changes in the composition of marine plankton and elevated carbon dioxide levels in the atmosphere. This study is based on high‐resolution sampling of two stratigraphic successions, located in Valdorbia (Umbria–Marche Apennines) and Monte Mangart (Julian Alps), Italy, which represent expressions of the Toarcian oceanic anoxic event in deep‐water pelagic sediments. These successions are characterized by the occurrence of black shales showing relatively low total organic carbon concentrations (compared with coeval strata in Northern Europe), generally < 2%, and low hydrogen indices. On this basis, they are similar to other Toarcian black shales described from the Tethyan region. The positive and negative carbon‐isotope records from the two localities permit a high‐resolution correlation such that ammonite biostratigraphy information from Valdorbia can be transferred to those parts of the Monte Mangart section that lack these fossils. Spectral analyses of δ13Corg values and of CaCO3 percentages from the sedimentary records of both the Valdorbia and Monte Mangart sections reveal a strong cyclic pattern, best interpreted as an eccentricity signal which hence implies a duration of ca 500 kyr for the negative carbon‐isotope excursion. Based on the carbon‐isotope curves obtained, the high‐resolution correlation between the Italian successions and a section in Yorkshire (Northern Europe) confirms the supposition that the apparent mismatch between the dating of the Toarcian oceanic anoxic event in the Boreal and Tethyan realms is an artefact of biostratigraphy.
Discrete lithofacies discrimination of Jurassic strata using Advanced Spaceborne Thermal Emission and Reflection Radiometer Data, Bighorn Basin, Wyoming, USA
Authors: STRASEN JAMES L. , CHAFETZ HENRY S. , KHAN SHUHAB
Published: February 2009

An investigation of the carbonate sedimentology of the Middle Jurassic strata, Bighorn Basin, North‐central Wyoming, proved to be an ideal test of new spaceborne remote sensing platforms with high spatial and spectral resolution. To evaluate these new tools, it was necessary to: (i) map the occurrence of different lithofacies in at least a part of the area to establish ground truth; (ii) establish the reflectance characteristics of the different lithofacies using portable, on the ground, systems; (iii) produce maps based on spaceborne remote sensing data that discriminate between the lithological types found in the field; and, lastly, (iv) evaluate the discrimination by visiting previously unvisited areas in which these rock types are predicted to occur based upon their remote signatures. Field checking of previously unvisited outcrops demonstrated whether these remote sensing techniques allowed distinction of different lithological outcrops. This investigation determined that the new spaceborne tools could distinguish between outcrops of sparse silty oomicrites from sorted biosparites as small as 10 m by 10 m. In a study area as large as the Bighorn Basin, the ability to identify very small outcrops with a predicted lithology a priori proved to be a major advantage in maximizing field effort.
Autocyclic behaviour of fan deltas: an analogue experimental study
Authors: VAN DIJK MAURITS , POSTMA GEORGE , KLEINHANS MAARTEN G.
Published: February 2009

Fan deltas are excellent recorders of fan‐building processes because of their high sedimentation rate, particularly in tectonically active settings. Although previous research focuses mainly on allogenic controls, there is clear evidence for autogenically produced storage and release of sediment by flume and numerical modelling that demands further definition of characteristics and significance of autogenically forced facies and stratigraphy. Analogue experiments were performed on fan deltas with constant extrinsic variables (discharge, sediment supply, sea‐level and basin relief) to demonstrate that fan‐delta evolution consists of prominent cyclic alternations of channellized flow and sheet flow. The channellized flow is initiated by slope‐induced scouring and subsequent headward erosion to form a channel that connected with the valley, while the removed sediment is deposited in a rapidly prograding delta lobe. The resulting decrease in channel gradient causes a reduction in flow strength, mouth‐bar formation, flow bifurcation and progressive backfilling of the channel. In the final stage of channel filling, sheet flow coexists for a while with channellized flow (semi‐confined flow), although in cycle 1 this phase of semi‐confined flow was absent. Subsequent autocyclic incisions are very similar in morphology and gradient. However, they erode deeper into the delta plain and, as a result, take more time to backfill. The duration of the semi‐confined flow increases with each subsequent cycle. During the period of sheet flow, the delta plain aggrades up to the ‘critical’ gradient required for the initiation of autocyclic incision. This critical gradient is dependent on the sediment transport capacity, defined by the input conditions. These autogenic cycles of erosion and aggradation confirm earlier findings that storage and release of sediment and associated slope variation play an important role in fan‐delta evolution. The erosional surfaces produced by the autocyclic incisions are well‐preserved by the backfilling process in the deposits of the fan deltas. These erosional surfaces can easily be misinterpreted as climate, sea‐level or tectonically produced bounding surfaces.
Molecular signatures of apomictic and sexual ovules in the Boechera holboellii complex
Authors: Sharbel Timothy F. , Voigt Marie‐Luise , Corral José María , Thiel Thomas , Varshney Alok , Kumlehn Jochen , Vogel Heiko , Rotter Björn
Published: February 2009

Apomixis, a natural form of asexual seed production in plants, has evolved independently in various taxa, and represents an important potential technology for agriculture. The switch to apomixis is based on de‐regulation of developmental pathways originally leading to sexual seed formation. Hybridization and polyploidy, both typical characteristics of asexual plants and animals, are mechanisms that could trigger de‐regulation. Here we show that up‐regulation of alleles in apomeiotic ovules is mediated by genomic duplication, heterochrony and the residual effects of ancient hybridization in diploid apomicts of the Boechera holboellii complex. Using SuperSAGE, we have identified over 4000 differentially expressed mRNA tags between micro‐dissected ovules from two diploid sexual (Boechera stricta and B. holboellii) and two diploid apomictic (Boechera divaricarpa) accessions. Pairwise sequence comparisons between tags enabled identification of allelic variants of the same loci. Up‐regulated candidate apomeiosis alleles consistently have more than three related alleles, thus demonstrating transcription from duplicated loci. A further 543 alleles were heterochronically expressed between sexual and apomeiotic ovules at developmental stages 2‐II to 2‐IV. Intriguingly, 69 B. holboellii specific alleles were preferentially up‐regulated in apomeiotic ovules, thus showing a remnant‘parent of origin’ effect stemming from the Pleistocene origin of the hybrid B. divaricarpa from taxa related to B. holboellii and B. stricta. These data implicate polyploid gene dosage in the expression of asexual seed formation, and support hypotheses of de‐regulation of the sexual pathway. The observed ‘parent of origin’ effect suggests that the genomic memory of hybridization has somehow been maintained after hundreds, if not thousands, of asexual generations.
Fusarium oxysporum hijacks COI1‐mediated jasmonate signaling to promote disease development in Arabidopsis
Authors: Thatcher Louise F. , Manners John M. , Kazan Kemal
Published: February 2009

Although defense responses mediated by the plant oxylipin jasmonic acid (JA) are often necessary for resistance against pathogens with necrotrophic lifestyles, in this report we demonstrate that jasmonate signaling mediated through COI1 in Arabidopsis thaliana is responsible for susceptibility to wilt disease caused by the root‐infecting fungal pathogen Fusarium oxysporum. Despite compromised JA‐dependent defense responses, the JA perception mutant coronatine insensitive 1 (coi1), but not JA biosynthesis mutants, exhibited a high level of resistance to wilt disease caused by F. oxysporum. This response was independent from salicylic acid‐dependent defenses, as coi1/NahG plants showed similar disease resistance to coi1 plants. Inoculation of reciprocal grafts made between coi1 and wild‐type plants revealed that coi1‐mediated resistance occurred primarily through the coi1 rootstock tissues. Furthermore, microscopy and quantification of fungal DNA during infection indicated that coi1‐mediated resistance was not associated with reduced fungal penetration and colonization until a late stage of infection, when leaf necrosis was highly developed in wild‐type plants. In contrast to wild‐type leaves, coi1 leaves showed no necrosis following the application of F. oxysporum culture filtrate, and showed reduced expression of senescence‐associated genes during disease development, suggesting that coi1 resistance is most likely achieved through the inhibition of F. oxysporum‐incited lesion development and plant senescence. Together, our results indicate that F. oxysporum hijacks non‐defensive aspects of the JA‐signaling pathway to cause wilt‐disease symptoms that lead to plant death in Arabidopsis.
No decrease in annual risk of tuberculosis infection in endemic area in Cape Town, South Africa
Authors: Kritzinger Fiona E. , Den Boon Saskia , Verver Suzanne , Enarson Donald A. , Lombard Carl J. , Borgdorff Martien W. , Gie Robert P. , Beyers Nulda
Published: February 2009

Objective  To estimate the change in annual risk of tuberculosis infection (ARTI) in two neighbouring urban communities of Cape Town, South Africa with an HIV prevalence of approximately 2%, and to compare ARTI with notification rates and treatment outcomes in the tuberculosis (TB) programme.

Methods  In 1998–1999 and 2005, tuberculin skin test surveys were conducted to measure the prevalence of Mycobacterium tuberculosis infection and to calculate the ARTI. All 6 to 9‐year‐old children from all primary schools were included in the survey. Notification rates and treatment outcomes were obtained from the TB register.

Results  A total of 2067 children participated in the survey from 1998 to 1999 and a total of 1954 in 2005. Based on a tuberculin skin test cut‐off point of 10 mm, the ARTI was 3.7% (3.4–4.0%) in the 1998–1999 survey and 4.1% (3.8–4.5%) in 2005. The notification rate for pulmonary TB increased significantly from 646 per 100 000 in 1998 to 784 per 100 000 in 2002. In Ravensmead, there was no significant change in ARTI [first survey: 3.5% (3.1–3.9%), second survey: 3.2% (2.9–3.6%)], but in Uitsig the ARTI increased significantly from 4.1% (3.6–4.6%) to 5.8% (5.2–6.5%). The difference in ARTI between the two areas was associated with differences in reported case rates and the proportion of previously treated cases.

Conclusion  Tuberculosis transmission remains very high in these two communities and control measures to date have failed. Additional measures to control TB are needed.
Malignant glomus tumour in a German shepherd dog
Authors: Park Chun‐Ho , Kozima Daisuke , Tsuzuki Naoto , Ishi Yoshifumi , Oyamada Toshifumi
Published: February 2009

A solitary mass, 2.5 cm × 2.5 cm × 2.0 cm in size, was found in the subcutis adjacent to the ischial tuberosity of an 8‐year‐old male German shepherd dog. The excised mass was not encapsulated and the cut surface was solid, grey–white and had multiple red areas suggestive of haemorrhage. Microscopically, the neoplasm was composed of epithelioid cells interspersed with venous vessels of various sizes and irregular bundles of spindle‐shaped tumour cells with mucinous stroma. The nuclei showed anisokaryosis and many mitotic figures were noted. Immunohistochemically, the majority of tumour cells were positive for alpha smooth‐muscle actin and vimentin, but negative for cytokeratin, desmin, S‐100 and factor VIII‐related antigen. Ultrastructurally, the tumour cells contained irregular nuclei, few mitochondria, few rough endoplasmic reticulum, cytoplasmic myofilaments of intermediate density, pinocytotic vesicles and distinct basal lamina. Based on these pathological findings, the diagnosis was malignant glomus tumour.
Greater impact of extreme drought on photosynthesis of grasslands exposed to a warmer climate in spite of acclimation
Authors: Zavalloni Costanza , Gielen Birgit , De Boeck Hans J. , Lemmens Catherine M.H.M. , Ceulemans Reinhart , Nijs Ivan
Published: February 2009

In view of the projected increase in the frequency of extreme events during this century, we investigated the impact of a drought extreme on leaf ecophysiological parameters and carbon isotope composition (δ13C) of grassland communities with species richness (S) of one, three or nine species. The communities, grown for 3 years at either ambient air temperatures (ambient Tair) or ambient Tair + 3°C (elevated Tair), were additionally subjected to an imposed drought by withholding water for 24 days. During the previous 3 years equal precipitation was applied in both temperature treatments, thus communities at elevated Tair had experienced more frequent, mild droughts. However, it was unknown whether this resulted in a higher resistance for facing extreme droughts. At similar soil matric potentials stomatal conductance (gs) and transpiration (Tr) were higher at elevated than ambient Tair, indicating acclimation to lower soil water content. Despite the stomatal acclimation observed, plants in elevated Tair showed a lower resistance to the drought extreme as indicated by their lower photosynthetic rate (Amax), gs and Tr during the entire duration of the drought extreme. Lower values for Amax, Tr and gs were also recorded in species at S = 3 as compared with species at S = 1 for both temperature treatments, but no further differences with S = 9 suggesting that stress was not alleviated at higher S‐levels. The discrimination of 13C was poorly correlated with measurements of instantaneous leaf water‐use efficiency (Amax/Tr) and, with this time scale and sampling method, it was not possible to detect any potential change in plant water‐use efficiency using leaf δ13C.
Seasonal variations in UV‐absorbing compounds and physiological characteristics in the aquatic liverwort Jungermannia exsertifolia subsp. cordifolia over a 3‐year period
Authors: Núñez‐Olivera Encarnación , Otero Saúl , Tomás Rafael , Martínez‐Abaigar Javier
Published: February 2009

Temporal physiological variations in relation to environmental factors, in particular to ultraviolet (UV) radiation, have been studied in bryophytes from circumpolar latitudes, but not in mid‐latitudes with longer growing seasons. In addition, seasonal and interannual changes in individual UV‐absorbing compounds (UVAC) have not been previously studied in bryophytes. To fill these gaps, samples of the aquatic liverwort Jungermannia exsertifolia subsp. cordifolia were collected on a monthly basis during 3 years from a mountain stream in northern Spain. Sclerophylly index, chlorophyll fluorescence, DNA damage, the bulk UV absorbance of methanolic extracts and the concentration of five UVAC (hydroxycinnamic acid derivatives) were measured. Interannual changes were little marked, probably because the 3 years studied were environmentally similar. In summer‐autumn, with respect to seasonal variations, newly grown tender young shoots with high Fv/Fm accumulated higher amounts of several hydroxycinnamic acid derivatives than in winter‐spring. DNA damage was not detected in any of the samples analyzed. p‐Coumaroylmalic acid was the compound best associated with radiation changes, and the best model explaining UV‐B took into consideration the concentration of this compound and the ozone level. The specific effects of UV radiation could not be separated from the effects caused by other environmental factors, such as global radiation or temperature, because all these variables were correlated. However, indirect evidence strongly suggests that seasonal changes in bulk UV absorbance and p‐coumaroylmalic acid are mainly driven by UV radiation. This compound may be a promising physiological variable to be used for UV bioindication.
Regulation of AhFRO1, an Fe(III)‐chelate reductase of peanut, during iron deficiency stress and intercropping with maize
Authors: Ding Hong , Duan Lihong , Wu Huilan , Yang Rongxin , Ling Hongqing , Li Wen‐Xue , Zhang Fusuo
Published: February 2009

Iron deficiency‐induced chlorosis in peanut during anthesis was alleviated when peanut was intercropped with maize in field and pot experiments. Iron acquisition of graminaceous plants is characterized by the synthesis and secretion of the iron‐chelating phytosiderophores. Compared to the roots of monocropped maize, the roots of maize intercropped with peanut always secreted higher amounts of phytosiderophores during peanut anthesis. For non‐graminaceous plants, reduction of ferric to ferrous iron on the root surface is the rate‐limiting step for mobilizing iron from soil. The full‐length cDNA, AhFRO1, which is encoding an Fe(III)‐chelate reductase, was isolated from peanut. AhFRO1 expression in yeast conferred Fe(III)‐chelate reductase activity to the cells. Consistent with its function in iron uptake, AhFRO1 was determined to be a membrane protein by transient expression analysis. AhFRO1 mRNA accumulated under iron deficiency conditions. During pre‐anthesis, the Fe(III)‐chelate reductase activity and the transcript levels of AhFRO1 were similar in monocropped and intercropped peanut. When the iron deficiency‐induced chlorosis developed in the monocropped peanuts, both the Fe(III)‐chelate reductase activity of peanut and the transcript levels of AhFRO1 were higher in intercropped than in monocropped peanuts, which is consistent with the secretion of phytosiderophores by maize roots. We conclude that AhFRO1 in peanut and phytosiderophores from maize co‐operate to improve the iron nutrition of peanut when intercropped with maize.
Ethylene and salicylic acid control glutathione biosynthesis in ozone‐exposed Arabidopsis thaliana
Authors: Yoshida Seiji , Tamaoki Masanori , Ioki Motohide , Ogawa Daisuke , Sato Yuko , Aono Mitsuko , Kubo Akihiro , Saji Shoko , Saji Hikaru , Satoh Shinobu , Nakajima Nobuyoshi
Published: February 2009

Ozone produces reactive oxygen species and induces the synthesis of phytohormones, including ethylene and salicylic acid. These phytohormones act as signal molecules that enhance cell death in response to ozone exposure. However, some studies have shown that ethylene and salicylic acid can instead decrease the magnitude of ozone‐induced cell death. Therefore, we studied the defensive roles of ethylene and salicylic acid against ozone. Unlike the wild‐type, Col‐0, Arabidopsis mutants deficient in ethylene signaling (ein2) or salicylic acid biosynthesis (sid2) generated high levels of superoxide and exhibited visible leaf injury, indicating that ethylene and salicylic acid can reduce ozone damage. Macroarray analysis suggested that the ethylene and salicylic acid defects influenced glutathione (GSH) metabolism. Increases in the reduced form of GSH occurred in Col‐0 6 h after ozone exposure, but little GSH was detected in ein2 and sid2 mutants, suggesting that GSH levels were affected by ethylene or salicylic acid signaling. We performed gene expression analysis by real‐time polymerase chain reaction using genes involved in GSH metabolism. Induction of γ‐glutamylcysteine synthetase (GSH1), glutathione synthetase (GSH2), and glutathione reductase 1 (GR1) expression occurred normally in Col‐0, but at much lower levels in ein2 and sid2. Enzymatic activities of GSH1 and GSH2 in ein2 and sid2 were significantly lower than in Col‐0. Moreover, ozone‐induced leaf damage observed in ein2 and sid2 was mitigated by artificial elevation of GSH content. Our results suggest that ethylene and salicylic acid protect against ozone‐induced leaf injury by increasing de novo biosynthesis of GSH.
Physiological and proteomic responses of two contrasting Populus cathayana populations to drought stress
Authors: Xiao Xiangwen , Yang Fan , Zhang Sheng , Korpelainen Helena , Li Chunyang
Published: February 2009

The acclimation of plants to water deficit is the result of many different physiological and biochemical mechanisms. To gain a better understanding of drought stress acclimation and tolerance mechanisms in Populus cathayana Rehder, we carried out an integrated physiological and comparative proteomic analysis on the drought stress responses of two contrasting populations originating from wet and dry regions in western China. The plantlets were subjected to continuous drought stress by withholding soil water content at 25% of field capacity (FC) for 45 days, while the control treatments were kept at 100% FC. Drought stress significantly inhibited plant growth, decreased net photosynthetic rate and stomatal conductance of leaves, increased the relative electrolyte leakage and malondialdehyde (MDA) content, and, at the same time, accumulated soluble sugars and free proline in both populations tested. The population from the dry climate region exhibited stronger tolerance to drought stress compared with the wet climate population. The proteomic analyses resulted in the identification of 40 drought‐responsive proteins. The functional categories of these proteins include the regulation of transcription and translation, photosynthesis, cytoskeleton, secondary metabolism, HSPs/chaperones, redox homeostasis and defense response. The results suggest that poplars’ tolerance to drought stress relates to the control of reactive oxygen species (ROS) and to osmoprotective capacity. The differential regulation of some drought‐responsive proteins, such as HSPs and the enzymes related to redox homeostasis and regulation of secondary metabolism, plays an important role in poplars’ tolerance and acclimation to drought stress. In conclusion, acclimation to water deficit involves changes in cellular metabolism and the regulation of gene networks. The present study not only provides new insights into the mechanisms of acclimation and tolerance to drought stress in different poplar populations but also provides clues for improving poplars’ drought tolerance through breeding or genetic engineering.
Report of the Third International Workshop on Molecular Blood Group Genotyping
Authors: Daniels G. , Van Der Schoot C. E. , Gassner C. , Olsson M. L.
Published: February 2009

The Third International Society of Blood Transfusion Workshop on Molecular Blood Group Genotyping was held in 2008, with a feedback meeting at the International Society of Blood Transfusion Congress in Macao SAR, China. Thirty‐three laboratories participated, eight less than in 2006. Six samples were distributed: sample 1 representing DNA from a sample referred because of abnormal serological results in D testing; samples 2 and 3 from transfusion‐dependent patients for testing for all clinically important polymorphisms; sample 4 a mixture of two DNA samples designed to simulate a chimera, referred because of abnormal serological results in donor testing; and samples 5 and 6 plasma samples from RhD‐negative pregnant women, for fetal RhD testing (only tested by 17 laboratories). For samples 1–3, 24 of 33 laboratories obtained completely correct results. For sample 4, the ability to detect the minority DNA population was partly dependent on method. Of the 17 laboratories that received samples 5 and 6, 13 reported correct results on both samples. Overall a small improvement from previous workshops was noted, but there is still room for improvement. The main conclusion for the 2006 workshop can be reiterated: with greater care and attention to detail, very high standards could be set for molecular blood group genotyping.
Consequences of near and far between‐population crosses for offspring fitness in a rare herb
Authors: Raabová J. , Münzbergová Z. , Fischer M.
Published: February 2009

Crosses between plants from different populations may result in heterosis or outbreeding depression. However, despite its importance for conservation, little is known about the spatial scale over which these effects may arise. To investigate the consequences of between‐population crosses at two distinct spatial scales, we conducted reciprocal crosses between four populations from two regions in the rare perennial herb Aster amellus. We assessed seed set and offspring fitness in a common garden experiment. Overall, between‐population crosses within regions (10 km) resulted in 8% lower seed set than within‐population crosses, while between‐region crosses (70 km) resulted in 17% higher seed set than within‐population crosses. Moreover, offspring from between‐population crosses produced 18% more flower heads than offspring from within‐population crosses. We conclude that hybridisation between A. amellus plants from different populations did not lead to immediate outbreeding depression and, thus, could represent a valid conservation option to increase genetic diversity. Moreover, our results suggest that the distance between populations affects the outputs of between‐population crosses and therefore needs to be taken into account when promoting gene flow between populations.
PROTECTIVE EFFECTS OF ICARIIN ON COGNITIVE DEFICITS INDUCED BY CHRONIC CEREBRAL HYPOPERFUSION IN RATS
Authors: Xu Rui‐Xia , Wu Qin , Luo Yong , Gong Qi‐Hai , Yu Li‐Mei , Huang Xie‐Nan , Sun An‐Sheng , Shi Jing‐Shan
Published: February 2009

Icariin is a major constituent of flavonoids derived from the Chinese medicinal herb Epimedium revicornum Maxim. The aim of the present study was to investigate whether icariin has protective effects on learning ability and memory in a rat model of chronic cerebral hypoperfusion.

Chronic cerebral hypoperfusion was induced by permanent ligation of the common carotid artery in Wistar rats for 4 months. One month after permanent artery occlusion, rats were adminitered icariin at doses of 0, 30, 60 or 120 mg/kg per day, p.o., for 3 months. Neurobehavioural and neurobiochemical parameters were examined to evaluate the effects of icariin on cognitive deficits induced by chronic cerebral hypoperfusion.

The Morris water maze test revealed that learning ability and memory were severely impaired in untreated rats, but were significantly improved in icariin‐treated rats. Icariin treatment also ameliorated chronic cerebral hypoperfusion‐induced oxidative stress in the brain, as evidenced by reduced malondialdehyde formation and maintained superoxide dismutase activity. In addition, the decreased hippocampal levels of acetylcholine, acetylcholinesterase and choline acetyltransferase associated with chronic cerebral hypoperfusion were significantly prevented by icariin treatment.

In conclusion, icariin protects against cognitive deficits induced by chronic cerebral hypoperfusion in rats. These effects appear to be mediated through its anti‐oxidant effects, as well as its effects on the circulatory and cholinergic systems.
HERG‐F463L POTASSIUM CHANNELS LINKED TO LONG QT SYNDROME REDUCE IKr CURRENT BY A TRAFFICKING‐DEFICIENT MECHANISM
Authors: Yang Hai‐Tao , Sun Chao‐Feng , Cui Chang‐Cong , Xue Xiao‐Lin , Zhang Ai‐Feng , Li Hong‐Bing , Wang Dong‐Qi , Shu Juan
Published: February 2009

Congenital long QT syndrome (LQTS) is a genetically heterogeneous disease. The aim of the present study was to identify the gene mutation in a Chinese family with LQTS and investigate the functional changes associated with the mutation.

Polymerase chain reaction and DNA sequencing were used to screen for the KCNH2 mutation in the proband. A mutant F463L HERG channel was expressed in HEK293 cells using a lipofectamine method. The IKr current was recorded using the whole‐cell voltage clamp technique. Expression of HERG protein was detected by western blotting and the subcellular location of HERG channels in cell was analysed by confocal microscopy.

The novel heterozygous missense mutation F463L in KCNH2 was detected. We found that the F463L mutation did not lead to any expression of detectable IKr current, which was consistent with western blotting analysis indicating that the F463L mutation only expressed a band at 135 kDa. When coexpressed with wild‐type HERG, F463L HERG exhibited strong dominant‐negative current suppression, resulting in a decrease in IKr current density, and induced a positive shift in the voltage dependence of activation, as well as interference with trafficking of wild‐type channel protein. The processing of the F463L channels was partly corrected in cells incubated in E4031. In addition, confocal microscopy demonstrated that F463L subunits could be inserted into the cell membrane when forming heteromultimeric channels with wild‐type channel subunits.

The results of the present study suggest that the F463L mutation leads to loss of function in HERG through a dominant‐negative effect caused by impaired trafficking of the channel.
MEASUREMENT OF PULMONARY FLOW RESERVE IN HIGHER PRIMATES
Authors: Ilsar Rahn , Chawantanpipat Chirapan , Chan Kim H , Waugh Richard , Hennessy Annemarie , Celermajer David S , Ng Martin KC
Published: February 2009

There are currently limited diagnostic methods for assessing the integrity of the pulmonary microvasculature. We hypothesized that a novel, invasively determined physiological index of ‘pulmonary flow reserve’ (PFR = maximal hyperaemic pulmonary blood flow divided by basal pulmonary flow) may facilitate microvascular assessment in the lung. Therefore, we developed a baboon model in which to: (i) validate the use of Doppler flow velocity for PFR assessment; (ii) define the optimal drug and dose regimen for attainment of maximal pulmonary hyperaemia; and (iii) demonstrate the feasibility of measuring PFR in healthy higher primates.

Doppler sensor guidewires were placed in segmental pulmonary arteries of 11 ketamine‐anaesthetized baboons. Vessel diameter, flow velocity and haemodynamics were recorded before and after direct intrapulmonary artery administration of saline, adenosine (50–500 µg/kg per min) and papaverine (3–60 mg), enabling calculation of PFR.

Saline (either bolus injection or infusion) did not alter vessel diameter or flow velocity (P > 0.1), validating local drug administration. Both adenosine and papaverine induced dose‐dependent increases in flow velocity from baseline (from 22.5 ± 2.3 to 32.7 ± 4.8 cm/s for 400–500 µg/kg per min adenosine; and from 23.9 ± 1.1 to 34.6 ± 4.0 cm/s for 24 mg papaverine; both P < 0.0001), without affecting pulmonary artery pressure or vessel diameter (P > 0.3). Healthy primate PFR values were 1.35 ± 0.10 and 1.39 ± 0.10 using 200 µg/kg per min adenosine and 24 mg papaverine, respectively (P > 0.8).

In conclusion, pulmonary flow reserve in higher primates can be assessed using Doppler sensor guidewire and either adenosine or papaverine as microvascular hyperaemic agents. Measurements of PFR may facilitate pulmonary microvascular assessments.
Effectiveness of prokinetic agents against diseases external to the gastrointestinal tract
Authors: Hiyama Toru , Yoshihara Masaharu , Tanaka Shinji , Haruma Ken , Chayama Kazuaki
Published: February 2009

Prokinetic agents are effective not only for disease of the gastrointestinal (GI) tract but also for those external to the GI tract such as the central nervous system, and the respiratory, urologic, and metabolic organs. This article reviews the effectiveness of prokinetic agents against diseases external to the GI tract. Studies were identified by computerized and manual searches of the available literature. A Medline search was performed (1975–July, 2008) using the following medical subject headings: prokinetic agent, metoclopramide, domperidone, trimebutine, cisapride, itopride, mosapride, tegaserod, and human. The identified diseases for which prokinetic agents may be effective are various: bronchial asthma, chronic cough, hiccup, spontaneous bacterial peritonitis, cholelithiasis, diabetes mellitus, acute migraine, Parkinson's disease, anorexia nervosa, Tourette's disorder, urologic sequelae of spinal cord injury and of radical hysterectomy for cervical cancer, laryngeal dysfunction and so on. These agents are also useful for prevention of aspiration pneumonia during anesthesia, and in tube‐fed patients. Prokinetic agents should be a valuable addition to our currently limited pharmacological armamentarium not only for functional bowel disease, but also for diseases external to the GI tract.
One‐hour fast for water and six‐hour fast for solids prior to endoscopy provides good endoscopic vision and results in minimum patient discomfort
Authors: De Silva Arjuna P , Amarasiri Lakmali , Liyanage Manohari N , Kottachchi Dulani , Dassanayake Anuradha S , De Silva H Janaka
Published: February 2009

Background and Aim:  Current guidelines for upper gastrointestinal endoscopy (UGIE) advise at least 6–8 h fasting for solids and 4‐h fasting for liquids. We aimed to determine whether a 6‐h fast for solids and one‐hour fast for water prior to UGIE gives good endoscopic vision and less patient discomfort.

Methods:  128 patients referred for UGIE were given a standard meal 6 h before endoscopy, and then randomized to either nil by mouth for 6 h (group A, n = 65) or allowed to drink water for up to one hour prior to endoscopy (group B, n = 63). Before endoscopy patients were requested to indicate discomfort due to fasting on a visual analog scale. Fluid in the gastric fundus was aspirated, when present, for volume and pH measurements, and endoscopic vision was graded.

Results:  53 patients in group A and 43 patients in group B completed the study. Discomfort was significantly lower in group B than group A (P < 0.0001). Endoscopic vision was good in all 53 patients in group A and 40 in group B, and average in 3 patients in group B. Fluid in the gastric fundus was noted in 11 patients in group A and 16 in group B, but there were no significant differences in volume or pH between groups. There were no complications attributable to endoscopy in either group.

Conclusions:  A 6‐h fast for solids and a 1‐h fast for water prior to UGIE gives good endoscopic vision, and causes minimum patient discomfort.
Medical practice, procedure manuals and the standardisation of hospital death
Authors: Hadders Hans
Published: February 2009

This paper examines how death is managed in a larger regional hospital within the Norwegian health‐care. The central focus of my paper concerns variations in how healthcare personnel enact death and handle the dead patient. Over several decades, modern standardised hospital death has come under critique in the western world. Such critique has resulted in changes in the standardisation of hospital deaths within Norwegian health‐care. In the wake of the hospice movement and with greater focus on palliative care, doors have gradually been opened and relatives of the deceased are now more often invited to participate. I explore how the medical practice around death along with the procedure manual of post‐mortem care at Trondheim University Hospital has changed. I argue that in the late‐modern context, standardisation of hospital death is a multidimensional affair, embedded in a far more comprehensive framework than the depersonalised medico‐legal. In the late‐modern Norwegian hospital, interdisciplinary negotiation and co‐operation has allowed a number of different agendas to co‐exist, without any ensuing loss of the medical power holder's authority to broker death. I follow Mol's notion of praxiographic orientation of the actor–network approach while exploring this medical practice.
Struggling to adapt: caring for older persons while under threat of organizational change and termination notice
Authors: Fläckman Birgitta , Hansebo Görel , Kihlgren Annica
Published: February 2009

Organizational changes are common in elder care today. Such changes affect caregivers, who are essential to providing good quality care. The aim of the present study was to illuminate caregivers’ experiences of working in elder care while under threat of organizational change and termination notice. Qualitative content analysis was used to examine interview data from 11 caregivers. Interviews were conducted at three occasions during a two‐year period. The findings show a transition in their experiences from ‘having a professional identity and self‐confidence’, to ‘being a professional in a threatening situation caused by someone else’ and to ‘struggling to adapt to a changed working environment as a person and a professional’. The caregivers experienced a loss of pride and satisfaction. Previous literature indicates that this may have consequences for the quality of care and that employees may be at risk of negative health effects. However, the caregivers continued to struggle, doing their best to complete their duties. The study has implications for high‐level decision‐makers, managers and caregivers in similar work–life situations in that it deals with factors that facilitate or impede similar transitions.
‘Having a laugh’: masculinities, health and humour
Authors: Williams Robert
Published: February 2009

There is longstanding interest within anthropology and sociology in the meaning of humour, but little research that examines humour within fathers’ health experiences. This paper specifically analyses fathers’ stories about humour shared with other men, and the links between gender and health, in order to identify the implications for health‐care and future research. Findings indicate that humour is an important aspect of fathers’ experiences of social connectedness with other men. Indeed, for African‐Caribbean fathers specifically, humour was an important aspect of their relationships with other ethnic minority men. Humour was also used to objectify, humiliate or ridicule others, for example in the form of sexualised or racist humour. However, fathers’ stories were also mediated by masculinities, it that masculinities enabled fathers to avoid disclosure of vulnerability regarding health experiences to others. The links between masculinities and health, the implications for interviewing fathers, and the implications for future research and healthcare practice with fathers are also discussed.
Propofol for tracheal intubation in children anesthetized with sevoflurane: a dose–response study
Authors: LERMAN JERROLD , HOULE TIMOTHY T. , MATTHEWS BENJAMIN T. , HOUCK JAMES , BURROWS FREDERICK A.
Published: February 2009

Background:  Tracheal intubation during sevoflurane induction is frequently facilitated with i.v. propofol. We designed a dose–response study to evaluate the intubating conditions, and the incidence and duration of apnea after i.v. propofol in children.

Methods/Materials:  Sixty healthy children were randomly assigned to 0, 0.5, 1, 2 or 3 mg·kg−1 i.v. propofol during sevoflurane/nitrous oxide anesthesia. Tracheal intubation was performed approximately 30 s after propofol by an anesthesiologist who was blind to the treatment. The anesthesiologist assessed the responses to laryngoscopy and intubation using a standardized scale. Incidence and duration of apnea after propofol as well as heart rate, and systolic blood pressure before and after laryngoscopy were recorded. Data were analyzed using one‐way and repeated measures anova, the Jonckheere–Terpstra test, and logistic regression, with P < 0.05 accepted.

Results:  The laryngoscopy score after 3 mg·kg−1 propofol was less than that after 0 mg·kg−1 (P < 0.01) and 0.5 mg·kg−1 (P < 0.05). Incidence of apnea after propofol 3 mg·kg−1, 8/10, was greater than after 0 mg·kg−1, 3/14 (P < 0.011) and 0.5 mg·kg−1, 3/12 (P < 0.03). Duration of apnea after 3 mg·kg−1 was greater than after 0 and 0.5 mg·kg−1 (P < 0.01). The risk of apnea increased 1.83 fold for each 1 mg·kg−1 dose increase in propofol (P < 0.01). Mean heart rate and systolic pressure decreased with the main effect, time.

Conclusion:  During sevoflurane/nitrous oxide anesthesia, propofol 3 mg·kg−1 provides superior intubating conditions with an increased incidence of and prolonged apnea compared with 0 and 0.5 mg·kg−1.
Detection and expression of the phosphonate transporter gene phnD in marine and freshwater picocyanobacteria
Authors: Ilikchyan Irina N. , McKay R. Michael L. , Zehr Jonathan P. , Dyhrman Sonya T. , Bullerjahn George S.
Published: February 2009

We describe a PCR‐based assay designed to detect expression of the phosphonate assimilation gene phnD from picocyanobacteria. The phnD gene encodes the phosphonate binding protein of the ABC‐type phosphonate transporter, present in many of the picocyanobacterial genome sequences. Detection of phnD expression can indicate a capacity of picoplankton to utilize phosphonates, a refractory form of phosphorus that can represent 25% of the high‐molecular‐weight dissolved organic phosphorus pool in marine systems. Primer sets were designed to specifically amplify phnD sequences from marine and freshwater Synechococcus spp., Prochlorococcus spp. and environmental samples from the ocean and Laurentian Great Lakes. Quantitative RT‐PCR from cultured marine Synechococcus sp. strain WH8102 and freshwater Synechococcus sp. ARC‐21 demonstrated induction of phnD expression in P‐deficient media, suggesting that phn genes are regulated coordinately with genes under phoRB control. Last, RT‐PCR of environmental RNA samples from the Sargasso Sea and Pacific Ocean detected phnD expression from the endemic picocyanobacterial population. Synechococcus spp. phnD expression yielded a depth‐dependent pattern following gradients of P bioavailability. By contrast, the Prochlorococcus spp. primers revealed that in all samples tested, phnD expression was constitutive. The method described herein will allow future studies aimed at understanding the utilization of naturally occurring phoshonates in the ocean as well as monitoring the acquisition of synthetic phosphonate herbicides (e.g. glyphosate) by picocyanobacteria in freshwaters.
Willow wood sap promotes the density‐dependent pathogenesis of Brenneria salicis
Authors: Huvenne Hanneke , Messens Eric , Maes Martine
Published: February 2009

Brenneria salicis resides in symptomless willow (Salix spp.) and other tree species, but only willow trees develop watermark disease. To understand the conversion of B. salicis into a pathogen, its pathogenicity and differential growth in the various tree species are studied. Brenneria salicis was detected by plating and polymerase chain reaction‐based techniques. Cell wall degradation and quorum sensing (QS) were assayed as possible pathogenicity mechanisms in wood. Differences in B. salicis growth capacities were tested in wood sap of the trees. Watermark diseased willow wood contained high concentrations of B. salicis with QS‐induced cellulase activity. In the fall, wood sap of willow, and not of poplar and alder, promoted high density growth of B. salicis. In situ, B. salicis was the dominant bacterial type in willow wood during the fall and winter period. Willow sustains high densities of B. salicis at the time of leaf shedding. The cellulase in the immobilized wood sap has then a long‐lasting contact with the xylem cell wall. Timing of dormancy and subsequent winter conditions might interfere with sap composition, B. salicis density, activity and survival, and be the reason, at least partly, for the variable occurrence of the disease.