bluuebunny/crossref_metadata_2025 huggingface dataset:
Red blood cells, platelets and polymorphonuclear neutrophils of patients with sickle cell disease exhibit oxidative stress that can be ameliorated by antioxidants
Authors: Amer Johnny , Ghoti Hussam , Rachmilewitz Eliezer , Koren Ariel , Levin Carina , Fibach Eitan
Published: March 2006

Sickle cell disease (SCD) is basically a red blood cell (RBC) disorder characterised by sickling and haemolysis, but platelets and polymorphonuclear neutrophils (PMN) are also involved. Oxidative damage may play a role in the pathogenesis of SCD. Using flow cytometry, we measured oxidative‐state markers simultaneously in RBC, platelets and PMN obtained from 25 normal donors, nine homozygous (SS) patients and six SS/beta‐thalassaemia patients. Reactive oxygen species (ROS) and reduced glutathione (GSH) were measured following staining of blood samples with fluorescence probes and gating on specific subpopulations based on size and granularity. Ten‐ to 30‐fold higher ROS production and 20–50% lower GSH content were found in RBC, platelets and PMN from SCD patients versus those of their normal counterparts. This could in part account for the clinical manifestations, such as haemolysis, a hypercoagulable state, recurrent bacterial infections and vaso‐occlusive incidences, in SCD. We further showed that exposure of SCD samples to antioxidants, such as N‐acetyl‐cysteine, vitamin C and vitamin E, decreased their oxidative stress. These results suggest that antioxidant treatment of patients with SCD could reduce oxidative damage to RBC, PMN and platelets, thereby alleviating symptoms associated with their pathology. The flow cytometry techniques presented herein could assist in monitoring the efficacy of such treatment.
Pelagic ecology of a northern boundary current system: effects of upwelling on the production and distribution of sardine (Sardinops sagax), anchovy (Engraulis australis) and southern bluefin tuna (Thunnus maccoyii) in the Great Australian Bight
Authors: WARD TIM M. , MCLEAY LACHLAN J. , DIMMLICH WETJENS F. , ROGERS PAUL J. , MCCLATCHIE SAM , MATTHEWS ROGER , KÄMPF JOCHEN , VAN RUTH PAUL D.
Published: March 2006

Shelf waters of southern Australia support the world's only northern boundary current ecosystem. Although there are some indications of intense nitrate enrichment in the eastern Great Australian Bight (GAB) arising from upwelling of the Flinders Current, the biological consequences of these processes are poorly understood. We show that productivity in the eastern GAB is low during winter, but that coastal upwelling at several locations during the austral summer–autumn results in localized increases in surface chlorophyll a concentrations and downstream enhancement of zooplankton biomass. Sardine (Sardinops sagax) and anchovy (Engraulis australis) eggs and larvae are abundant and widely distributed in shelf waters of the eastern and central GAB during summer–autumn, with high densities of sardine eggs and larvae occurring in areas with high zooplankton biomass. Egg densities and distributions support previous evidence suggesting that the spawning biomass of sardine in the waters off South Australia is an order of magnitude higher than elsewhere in southern Australia. Sardine comprised >50% of the identified prey species of juvenile southern bluefin tuna (SBT, Thunnus maccoyii) collected during this study. Other studies have shown that the lipid content of sardine from the GAB is relatively high during summer and autumn. We suggest that juvenile SBT migrate into the eastern and central GAB during each summer–autumn to access the high densities of lipid‐rich sardines that are available in the region during the upwelling period. Levels of primary, secondary and fish production in the eastern GAB during summer–autumn are higher than those recorded in other parts of Australia, and within the lower portion of ranges observed during upwelling events in the productive eastern boundary current systems off California, Peru and southern Africa.
Caveolin‐1 is down‐regulated and inversely correlated with HER2 and EGFR expression status in invasive ductal carcinoma of the breast
Authors: Park S‐S , Kim J E , Kim Y A , Kim Y C , Kim S‐W
Published: March 2006

Aims : To evaluate the caveolin‐1 status of invasive ductal carcinoma and its correlation with other important parameters of breast carcinogenesis. Caveolin‐1, the main structural protein of caveolae, is involved in the regulation of several intracellular signalling pathways and also functions as a tumour suppressor in breast carcinogenesis.

Methods and results : One hundred and thirty cases of invasive ductal carcinomas with matched normal breast tissue were evaluated immunohistochemically for caveolin‐1 expression. Using a tissue microarray, caveolin‐1 expression was also correlated with the expression of other antigens such as eostrogen receptor, progesterone receptor, epidermal growth factor receptor (EGFR), HER2, β‐catenin, E‐cadherin, p53, Ki67 and with clinicopathological parameters. Immunohistochemical results showed strong expression of caveolin‐1 in all normal breast epithelial cells, but a reduction of caveolin‐1 expression in 56 cases (43.1%) of invasive ductal carcinoma. Furthermore, a statistically significant inverse correlation between caveolin‐1 and EGFR and HER2 was noted (P < 0.001).

Conclusions : Our results indicate a reduction in caveolin‐1 expression in invasive ductal carcinoma of the breast, which supports in vitro studies of its role as a tumour suppressor. Caveolin‐1 also shows an inverse correlation with EGFR and HER2, which fits with its function as a negative regulator of signal transduction.
Continuous response functions for quantifying the strength of edge effects
Authors: EWERS ROBERT M. , DIDHAM RAPHAEL K.
Published: March 2006

Ecological boundaries are a dominant feature of human‐modified landscapes and have been the subject of numerous empirical studies. Robust statistical methods for determining the strength of edge effects are a vital requirement for the effective management of species that are negatively affected by habitat boundaries in heavily fragmented landscapes, but development of such methods has been slow.

We define edge effects as being composed of two complementary, and statistically definable, components: magnitude (the degree of difference in response values between patch and matrix interiors) and extent (the distance over which the difference in response values can be detected).

We present a statistical approach to rigorously delineate edge‐effect magnitude and extent. Our approach adapts a form of the general logistic model to describe continuous response functions for any biotic or abiotic variable across ecological boundaries from the landscape matrix into focal patch habitats. The model describes sigmoid and unimodal response functions that have been both theoretically predicted and empirically demonstrated. We use the second derivatives of the functions as an objective means to calculate the magnitude and extent of edge effects, and present a bootstrap technique for calculating confidence intervals around these values.

Synthesis and applications. Our results show clearly that edge‐effect magnitude and extent are not necessarily correlated, and therefore provide quantitatively different, and complementary, information about the strength of edge effects. Both effect magnitude and extent can be easily used for cross‐study comparisons, either by directly comparing the absolute values of response variables in the patch and matrix, or by converting those values to a percentage change. Furthermore, this method provides a management tool for more accurately predicting the presence, spatial location and utilization of core habitat by species in fragmented landscapes.
Widespread gene flow and high genetic variability in populations of water volesArvicola terrestrisin patchy habitats
Authors: AARS J. , DALLAS J. F. , PIERTNEY S. B. , MARSHALL F. , GOW J. L. , TELFER S. , LAMBIN X.
Published: March 2006

Theory predicts that the impact of gene flow on the genetic structure of populations in patchy habitats depends on its scale and the demographic attributes of demes (e.g. local colony sizes and timing of reproduction), but empirical evidence is scarce. We inferred the impact of gene flow on genetic structure among populations of water volesArvicola terrestristhat differed in average colony sizes, population turnover and degree of patchiness. Colonies typically consisted of few reproducing adults and several juveniles. Twelve polymorphic microsatellite DNA loci were examined. Levels of individual genetic variability in all areas were high (HO= 0.69–0.78). Assignments of juveniles to parents revealed frequent dispersal over long distances. The populations showed negativeFISvalues among juveniles,FISvalues around zero among adults, highFSTvalues among colonies for juveniles, and moderate, often insignificant,FSTvalues for parents. We inferred that excess heterozygosity within colonies reflected the few individuals dispersing from a large area to form discrete breeding colonies. Thus pre‐breeding dispersal followed by rapid reproduction results in a seasonal increase in differentiation due to local family groups. Genetic variation was as high in low‐density populations in patchy habitats as in populations in continuous habitats used for comparison. In contrast to most theoretical predictions, we found that populations living in patchy habitats can maintain high levels of genetic variability when only a few adults contribute to breeding in each colony, when the variance of reproductive success among colonies is likely to be low, and when dispersal between colonies exceeds nearest‐neighbour distances.
INVITED REVIEW: What is a population? An empirical evaluation of some genetic methods for identifying the number of gene pools and their degree of connectivity
Authors: WAPLES ROBIN S. , GAGGIOTTI OSCAR
Published: March 2006

We review commonly used population definitions under both the ecological paradigm (which emphasizes demographic cohesion) and the evolutionary paradigm (which emphasizes reproductive cohesion) and find that none are truly operational. We suggest several quantitative criteria that might be used to determine when groups of individuals are different enough to be considered ‘populations’. Units for these criteria are migration rate (m) for the ecological paradigm and migrants per generation (Nm) for the evolutionary paradigm. These criteria are then evaluated by applying analytical methods to simulated genetic data for a finite island model. Under the standard parameter set that includes L = 20 High mutation (microsatellite‐like) loci and samples of S = 50 individuals from each of n = 4 subpopulations, power to detect departures from panmixia was very high (∼100%; P < 0.001) even with high gene flow (Nm = 25). A new method, comparing the number of correct population assignments with the random expectation, performed as well as a multilocus contingency test and warrants further consideration. Use of Low mutation (allozyme‐like) markers reduced power more than did halving S or L. Under the standard parameter set, power to detect restricted gene flow below a certain level X (H0: Nm < X) can also be high, provided that true Nm ≤ 0.5X. Developing the appropriate test criterion, however, requires assumptions about several key parameters that are difficult to estimate in most natural populations. Methods that cluster individuals without using a priori sampling information detected the true number of populations only under conditions of moderate or low gene flow (Nm ≤ 5), and power dropped sharply with smaller samples of loci and individuals. A simple algorithm based on a multilocus contingency test of allele frequencies in pairs of samples has high power to detect the true number of populations even with Nm = 25 but requires more rigorous statistical evaluation. The ecological paradigm remains challenging for evaluations using genetic markers, because the transition from demographic dependence to independence occurs in a region of high migration where genetic methods have relatively little power. Some recent theoretical developments and continued advances in computational power provide hope that this situation may change in the future.
Relationship between leaf emergence and optimum spray timing for leaf blotch (Rhynchosporium secalis) control on winter barley
Authors: Young C. S. , Thomas J. M. , Parker S. R. , Paveley N. D.
Published: March 2006

For wheat, the optimum time to apply fungicide to control disease on a given leaf layer is usually at, or shortly after, full leaf emergence. Data from field experiments on barley were used to investigate whether the same relationship was applicable to control of leaf blotch on barley. Replicated plots of winter barley were sown in the autumns of 1991, 1992 and 1993 at sites in southwest England with high risk of Rhynchosporium secalis infection. Single fungicide treatments at four doses (0·25, 0·5, 0·75 or 1·0 times the label rate) were applied at one of eight different spray times, starting in mid‐March in each year, with intervals of 10–11 days between spray timings. Disease was assessed every 10–11 days and area under the disease progress curve (AUDPC) values were used to construct fungicide dose by spray time response surfaces for each of the upper four leaves, for each year. Spray timings shortly before leaf emergence were found to minimize the AUDPC for each year and leaf layer, and also the effective dose (the dose required to achieve a specified level of control), similar to wheat. Fungicide treatments on barley were effective for a longer period before leaf emergence than afterwards, probably because treatments before emergence of the target leaf reduced inoculum production on leaves below. This partly explains why fungicides tend to be applied earlier in the growth of barley compared with wheat.
A high‐throughput glasshouse bioassay to detect crown rot resistance in wheat germplasm
Authors: Mitter V. , Zhang M. C. , Liu C. J. , Ghosh R. , Ghosh M. , Chakraborty S.
Published: March 2006

A high‐throughput and reliable seedling bioassay to screen wheat germplasm for crown rot resistance was developed. Single wheat seedlings were grown in square seedling punnets in a glasshouse and inoculated with a monoconidial Fusarium pseudograminearum isolate 10 days after emergence. The punnets were laid horizontally on their side and a 10‐µL inoculum droplet placed on the stem base. Seedlings were incubated at near‐saturated relative humidity, and crown rot severity was assessed 35 days after inoculation. Studies on the duration of incubation period, inoculum concentration and temperature were carried out to optimize these parameters. Seedling growth at 25/15(±5)°C in a glasshouse and 48‐h incubation at near‐saturated RH in darkness gave the best results. When crown rot resistance rankings of 16 Australian cultivars from the bioassay were compared with their field performance, Spearman's rank correlation coefficient was highly significant. This indicated that the seedling bioassay mimicked field resistance to crown rot in adult plants. A bootstrap resampling analysis showed little or no improvement in the coefficient of variation with an increasing number of replications, indicating a high level of precision and reproducibility. By detecting small but consistent differences in crown rot severity, the bioassay proved effective in large‐scale screening for partial resistance: already over 1400 wheat genotypes have been screened. The high degree of precision makes this an invaluable tool in the understanding of pathogen aggressiveness, host specialization and parasitic fitness.
Metabolomic approaches reveal that phosphatidic and phosphatidyl glycerol phospholipids are major discriminatory non‐polar metabolites in responses by Brachypodium distachyon to challenge by Magnaporthe grisea
Authors: William Allwood J. , Ellis David I. , Heald Jim K. , Goodacre Royston , Mur Luis A. J.
Published: March 2006

Metabolomic approaches were used to elucidate some key metabolite changes occurring during interactions of Magnaporthe grisea– the cause of rice blast disease – with an alternate host, Brachypodium distachyon. Fourier‐transform infrared (FT‐IR) spectroscopy provided a high‐throughput metabolic fingerprint of M. grisea interacting with the B. distachyon accessions ABR1 (susceptible) and ABR5 (resistant). Principal component–discriminant function analysis (PC‐DFA) allowed the differentiation between developing disease symptoms and host resistance. Alignment of projected ‘test‐set’ on to ‘training‐set’ data indicated that our experimental approach produced highly reproducible data. Examination of PC‐DFA loading plots indicated that fatty acids were one chemical group that discriminated between responses by ABR1 and ABR5 to M. grisea. To identify these, non‐polar extracts of M. grisea‐challenged B. distachyon were directly infused into an electrospray ionization mass spectrometer (ESI‐MS). PC‐DFA indicated that M. grisea‐challenged ABR1 and ABR5 were differentially clustered away from healthy material. Subtraction spectra and PC‐DFA loadings plots revealed discriminatory analytes (m/z) between each interaction and seven metabolites were subsequently identified as phospholipids (PLs) by ESI‐MS‐MS. Phosphatidyl glycerol (PG) PLs were suppressed during both resistant and susceptible responses. By contrast, different phosphatidic acid PLs either increased or were reduced during resistance or during disease development. This suggests considerable and differential PL processing of membrane lipids during each interaction which may be associated with the elaboration/suppression of defence mechanisms or developing disease symptoms.
Conservation and divergence of plant microRNA genes
Authors: Zhang Baohong , Pan Xiaoping , Cannon Charles H. , Cobb George P. , Anderson Todd A.
Published: March 2006

MicroRNA (miRNA) is one class of newly identified, small, non‐coding RNAs that play versatile and important roles in post‐transcriptional gene regulation. All miRNAs have similar secondary hairpin structures; many of these are evolutionarily conserved. This suggests a powerful approach to predict the existence of new miRNA orthologs or homologs in other species. We developed a comprehensive strategy to identify new miRNA homologs by mining the repository of available ESTs. A total of 481 miRNAs, belonging to 37 miRNA families in 71 different plant species, were identified from more than 6 million EST sequences in plants. The potential targets of the EST‐predicted miRNAs were also elucidated from the EST and protein databases, providing additional evidence for the real existence of these miRNAs in the given plant species. Some plant miRNAs were physically clustered together, suggesting that these miRNAs have similar gene expression patterns and are transcribed together as a polycistron, as observed among animal miRNAs. The uracil nucleotide is dominant in the first position of 5′ mature miRNAs. Our results indicate that many miRNA families are evolutionarily conserved across all major lineages of plants, including mosses, gymnosperms, monocots and eudicots. Additionally, the number of miRNAs discovered was directly related to the number of available ESTs and not to evolutionary relatedness to Arabidopsis thaliana, indicating that miRNAs are conserved and little phylogenetic signal exists in the presence or absence of these miRNAs. Regulation of gene expression by miRNAs appears to have existed at the earliest stages of plant evolution and has been tightly constrained (functionally) for more than 425 million years.
Transfusion‐transmitted hepatitis E in a ‘nonhyperendemic’ country
Authors: Boxall E. , Herborn A. , Kochethu G. , Pratt G. , Adams D. , Ijaz S. , Teo C.‐G.
Published: March 2006

summary.  Indigenous hepatitis E is increasingly recognized in developed countries, where it may be a zoonosis. We describe the first case of transfusion‐transmitted hepatitis E in the UK from a blood donor who had no history of recent travel abroad.

Follow‐up of the donor and recipients of the blood products was carried out using serological and molecular techniques.

Acute hepatitis E was transmitted to one of two recipients. The infected patient would have received a larger volume of the donor's plasma. HEV subgenomic sequences carried by the donor and recipient were identical.

This is the first case of post‐transfusion hepatitis E in the UK. Secondary transmission of hepatitis E indigenous to a nonhyperendemic country may occur by blood transfusion. It is important that blood donors inform the transfusion service of all post‐donation illnesses so that appropriate interventions can take place.
Immature platelet fraction measurement: a future guide to platelet transfusion requirement after haematopoietic stem cell transplantation
Authors: Briggs C. , Hart D. , Kunka S. , Oguni S. , Machin S. J.
Published: March 2006

summary.  The decision to prophylactically transfuse platelets is dependent on the platelet count, careful regular clinical assessment and agreed local protocol. The ability to predict when platelet recovery will occur should allow a more reasoned approach to platelet transfusion. An increase in reticulated platelets demonstrates impending platelet recovery. A new rapid automated method to assess reticulated platelets, the immature platelet fraction (IPF), is described, and its clinical utility assessed.

The IPF is identified by flow cytometry with the use of a nucleic acid specific dye in the reticulocyte channel on the Sysmex XE‐2100. Fifty healthy adult volunteers were used to establish the normal range. Patients where platelet marrow production or destruction might be abnormal were studied, and some patients followed serially during treatment. Thirty patients receiving cytotoxic chemotherapy were tested, and 13 of these patients followed serially. Fifteen patients post‐autologous or allogeneic transplant were followed daily for platelet count and IPF percentage to monitor platelet recovery.

The method demonstrates good reproducibility and stability. The recovery phase of thrombocytopenia in most chemotherapy and transplant patients was preceded by a rise in IPF percentage several days prior to platelet recovery. In particular, patients undergoing autologous transplantation (n = 8) using peripherally collected stem cells have a characteristic IPF percentage motif, with a rise one or two days prior to engraftment.

The automated IPF is a useful parameter in the clinical evaluation of the thrombocytopenic patient and has the potential to allow optimal transfusion of platelet concentrates.
Cost‐effectiveness of asthma control: an economic appraisal of the GOAL study
Authors: Briggs A. H. , Bousquet J. , Wallace M. V. , Busse W. W. , Clark T. J. H. , Pedersen S. E. , Bateman E. D. ,   the GOAL Investigators Group
Published: March 2006

Background:  The Gaining Optimal Asthma ControL (GOAL) study has shown the superiority of a combination of salmeterol/fluticasone propionate (SFC) compared with fluticasone propionate alone (FP) in terms of improving guideline defined asthma control.

Methods:  Clinical and economic data were taken from the GOAL study, supplemented with data on health related quality of life, in order to estimate the cost per quality adjusted life year (QALY) results for each of three strata (previously corticosteroid‐free, low‐ and moderate‐dose corticosteroid users). A series of statistical models of trial outcomes was used to construct cost effectiveness estimates across the strata of the multinational GOAL study including adjustment to the UK experience. Uncertainty was handled using the non‐parametric bootstrap. Cost‐effectiveness was compared with other treatments for chronic conditions.

Result:  Salmeterol/fluticasone propionate improved the proportion of patients achieving totally and well‐controlled weeks resulting in a similar QALY gain across the three strata of GOAL. Additional costs of treatment were greatest in stratum 1 and least in stratum 3, with some of the costs offset by reduced health care resource use. Cost‐effectiveness by stratum was £7600 (95% CI: £4800–10 700) per QALY gained for stratum 3; £11 000 (£8600–14 600) per QALY gained for stratum 2; and £13 700 (£11 000–18 300) per QALY gained for stratum 1.

Conclusion:  The GOAL study previously demonstrated the improvement in total control associated with the use of SFC compared with FP alone. This study suggests that this improvement in control is associated with cost‐per‐QALY figures that compare favourably with other uses of scarce health care resources.
The effect of salt chamber treatment on bronchial hyperresponsiveness in asthmatics
Authors: Hedman J. , Hugg T. , Sandell J. , Haahtela T.
Published: March 2006

Background:  Randomized controlled trials are needed to evaluate the effects of complementary treatments in asthma. This study assessed the effect of salt chamber treatment as an add‐on therapy to low to moderate inhaled steroid therapy in asthma patients with bronchial hyperresponsiveness (BHR).

Methods:  After a 2‐week baseline period, 32 asthma patients who exhibited BHR in the histamine inhalation challenge were randomized: 17 to 2‐week active treatment, during which salt was fed to the room by a salt generator, and 15 to placebo. The salt chamber treatment lasted 40 min and was administered five times a week.

Results:  Median provocative dose causing a decrease of 15% in Fev1 (PD15FEV4) increased significantly in the active group (P = 0.047) but not in the placebo group. The difference in changes between the active and placebo groups was significant (P = 0.02). Nine patients (56%) in the active group and two patients (17%) in the placebo group exhibited at least one doubling dose decrease in BHR (P = 0.040). Six patients (38%) in the active group and none in the placebo group became non‐hyperresponsive (P = 0.017). Neither the peak expiratory flow (PEF) values measured just before and after the treatment, nor FEV1 values measured before the histamine challenges, changed. The reduction in BHR was not caused by changes in the baseline lung function.

Conclusions:  Salt chamber treatment reduced bronchial hyperresponsiveness as an add‐on therapy in asthmatics with a low to moderate dose of inhaled steroids. The possibility that salt chamber treatment could serve as a complementary therapy to conventional medication cannot be excluded.
Clinical profiles of four patients with Rett syndrome carrying a novel exon 1 mutation or genomic rearrangement in the MECP2 gene
Authors: Bartholdi D , Klein A , Weissert M , Koenig N , Baumer A , Boltshauser E , Schinzel A , Berger W , Mátyás G
Published: March 2006

Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the X‐linked MECP2 gene encoding methyl CpG binding protein 2 (MeCP2). Recently, a new isoform of MeCP2 including exon 1 was identified. This new isoform is more abundantly expressed in brain than the isoform including exons 2–4. Very little is known about the phenotypes associated with mutations in exon 1 of MECP2 since only a limited number of RTT patients carrying such mutations have been identified so far. In this study, we screened a cohort of 20 girls with RTT for exon 1 mutations by sequencing and multiplex ligation‐dependent probe amplification (MLPA). We identified one girl with a novel exon 1 mutation (c.30delCinsGA) by sequencing and three with genomic rearrangements by MLPA. Comparison of the phenotypes showed that the girls carrying a mutation or rearrangement encompassing exon 1 were more severely affected than the girls with rearrangements not affecting exon 1.
Hemoglobin is essential for normal growth of Arabidopsis organs
Authors: Hebelstrup Kim Henrik , Hunt Peter , Dennis Elizabeth , Jensen Susie Bjerregaard , Jensen Erik Østergaard
Published: March 2006

In Arabidopsis thaliana, the class I hemoglobin AHb1 is transiently expressed in the hydathodes of leaves and in floral buds from young inflorescences. Nitric oxide (NO) accumulates to high levels in these organs when AHb1 is silenced, indicating an important role in metabolizing NO. AHb1‐silenced lines are viable but show a mutant phenotype affecting the regions where AHb1 is expressed. Arabidopsis lines with an insertional knockout or overexpression of AHb2, a class II 3‐on‐3 hemoglobin, were generated. Seedlings overexpressing AHb2 show enhanced survival of hypoxic stress. The AHb2 knockout lines develop normally. However, when AHb2 knockout is combined with AHb1 silencing, seedlings die at an early vegetative stage suggesting that the two 3‐on‐3 hemoglobins, AHb1 and AHb2, together play an essential role for normal development of Arabidopsis seedlings. In conclusion, these results suggests that 3‐on‐3 hemoglobins apart from a role in hypoxic stress play a general role under non‐stressed conditions where they are essential for normal development by controlling the level of NO which tends to accumulate in floral buds and leaf hydathodes of plants.
Expression of a maize δ‐type DNA polymerase during seed germination
Authors: García E. , Quiroz F. , Uchiyama Y. , Sakaguchi K. , Vázquez‐Ramos J. M.
Published: March 2006

We have cloned a 1563‐bp cDNA sequence from a reported 2072‐bp cDNA (including a fragment of the 3′ UTR region, accession number AY109773) corresponding to the carboxy half of maize DNA polymerase δ[Zea mays delta‐type DNA polymerase catalytic subunit (Zmpolδ); EC 2.7.7.7], and its sequence shows an identity of 95, 77 and 74% to rice, soybean and Arabidopsis enzymes, respectively, although identity is even higher if only the polδ‐defining domain sequences are considered. An important difference between the monocot and dicot enzymes is the presence in the former of Zn fingers apparently required for binding to the DNA polδ holoenzyme B subunit, which is absent in the dicot enzymes. Expression of Zmpolδ and protein levels during germination remain unchanged; however, polδ shows two peaks of activity, one during early germination, perhaps related to DNA repair processes and a second peak when DNA replication is already in progress, indicating that Zmpolδ is regulated at the post‐translational level. Zmpolδ has been found mainly in proliferative tissues in seeds and plantlets, although surprisingly, it is also present in seed endosperm, together with Zm proliferating cell nuclear antigen.
ABO histo‐blood group antigen expression on the graft endothelium long term after ABO‐compatible, non‐identical heart transplantation
Authors: Koestner Simon C. , Kappeler Andreas , Schaffner Thomas , Carrel Thierry P. , Mohacsi Paul J.
Published: March 2006

Abstract: We recently reported a complete change in the endothelial ABO histo‐blood group phenotype of a cardiac allograft long term after B to O mismatched transplantation. In the context of the current controversy on graft recolonization with recipient endothelial cells and its importance in the development of immunological unresponsiveness, we monitored the expression of endothelial ABH histo‐blood group antigens of 10 ABO‐compatible, non‐identical cardiac allografts over an observation period of at least 30 months. ABH antigens as well as markers for endothelial cells, erythrocytes and thrombocytes were investigated retrospectively by immunohistochemistry using monoclonal antibodies on sections of formalin‐fixed, paraffin‐embedded biopsies and were evaluated semi‐quantitatively by microscopy. In contrast to our earlier finding of the change in the endothelial ABO histo‐blood group phenotype long term after ABO‐ mismatched transplantation, we could not confirm this change in 10 compatible but non‐identical cases.
A series of surveys on assay for anti‐A/B antibody by Japanese ABO‐incompatible Transplantation Committee
Authors: Kobayashi Takaaki , Saito Kazuhide
Published: March 2006

Abstract: Background: The measurement of anti‐blood groups A and B antibody (Ab) titers is considered to be important in ABO‐incompatible transplantation. However, no standard method for quantitative determination has yet been established in Japan. Inter‐examiner variability was likely because Ab titer was determined mainly by visual observation. In order to assess inter‐institutional variation in the measurement of anti‐A/B Ab titer, a series of surveys was conducted by the Japanese ABO‐incompatible Transplantation Committee.

Methods and Results: In 2003, the first national survey was conducted. Serum samples from six healthy volunteers with blood groups A (n=2), B (n=2) and O (n=2) were sent to 29 institutions and anti‐A/B Ab titer was measured by their customary methods. Isohemagglutinin assay in serial‐doubling dilutions of serum using a test tube was widely used in all institutions. Inter‐institutional difference between maximum and minimum value reached as much as 32‐fold in immunoglobulin M (IgM) and 256‐fold in IgG. As detailed protocol for assay seemed to be different between institutions, we attempted to standardize the protocol based on the result of a questionnaire survey. In 2004, a second survey was conducted in the same manner as the previous one, except participation involved 38 institutions and the measurement was performed in a uniform way using a provisional standard protocol. Analysis of the survey revealed that intra‐institutional variation was reduced to below eightfold, except that several institutions showed a large difference from the mean titer and required some guidance.

Conclusions: A periodical quality control survey is considered necessary to improve the accuracy of measurement. Anti‐A/B Ab titer would provide useful information towards the prediction of rejection and the indication of treatments such as (double filtration) plasmapheresis, splenectomy, anti‐CD20 monoclonal antibody and intravenous immunoglobulin. Standardization of the assay for anti‐A/B Ab titer is essential for the fulfillment of a precise multicenter study, which will elucidate the significance of the measurement of anti‐A/B Ab titer.
The Anti‐microbial Activity of Electrolysed Oxidizing Water against Microorganisms relevant in Veterinary Medicine
Authors: Fenner D. C. , Bürge B. , Kayser H. P. , Wittenbrink M. M.
Published: March 2006

Standards of the German Association of Veterinary Medicine (DVG) for the evaluation of chemical disinfectants were used to assess the anti‐microbial efficacy of electrolysed oxidizing water (EOW). Enterococcus faecium, Mycobacterium avium subspecies avium, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans were exposed to anode EOW (pH, 3.0 ± 0.1; oxidation‐reduction potential (ORP), +1100 ± 50 mV; free chlorine, 400 ± 20 mg/l Cl2) and combined EOW (7 : 3 anode : cathode, v/v; pH, 8.3 ± 0.1; ORP, 930–950 mV; free chlorine, 271 ± 20 mg/l Cl2). In water of standardized hardness (WSH), all bacterial strains were completely inactivated by a 30 min exposure to maximum 10.0% anode EOW (∼40.0 mg/l Cl2) or 50.0% combined EOW (∼135.5 mg/l Cl2). The sensitivity ranking order for anode EOW to the bacterial test strains was P. mirabilis > S. aureus > M. avium ssp. avium > E. faecium > P. aeruginosa. P. mirabilis and S. aureus decreased to undetectable levels after 5 min of exposure to 7.5% anode EOW (∼30.0 mg/l Cl2). Candida albicans was completely inactivated by a 5‐min exposure to 5.0% anode EOW. Both, anode and combined EOW exhibited no anti‐microbial activities in standardized nutrient broth or after addition of 20.0% bovine serum to the WSH. Further research is necessary to evaluate the efficacy of EOW as a disinfectant under operating conditions in animal production facilities.
Prevalence of Thermophilic Campylobacter Infections in Humans, Chickens and Crows in Morogoro, Tanzania
Authors: Mdegela R. H. , Nonga H. E. , Ngowi H. A. , Kazwala R. R.
Published: March 2006

Prevalence of thermophilic Campylobacter infections in humans, chickens and crows was determined in a cross‐sectional study that was carried out in urban and rural areas of Morogoro region, Tanzania during the period of January 2003 to December 2004. A total of 632 human stool samples, 536 cloacal swabs from local and broiler chickens and 22 intestinal contents from crows were screened for presence of thermophilic campylobacters using Skirrow's protocol. Representative Campylobacter jejuni isolates from human and chicken samples were also analysed by polymerase chain reaction (PCR) as a definitive identification method. The overall prevalence of thermophilic campylobacters was 9.3% (95% CI: 7.2–11.9), 69.8% (95% CI: 65.7–73.6) and 72.7% (95% CI: 49.8–89.3) in humans, chickens and crows respectively. In humans, 59 thermophilic campylobacters were isolated of which 96.6% were C. jejuni and 3.4%Campylobacter coli. There was a significantly (P < 0.001) higher prevalence in young individuals (16%) than in adults (7%). Of 341 isolates from chickens, 91.2% were C. jejuni and 8.8% were C. coli. A significantly (P < 0.05) higher infection rate was observed in rural local chicken (76%) than in broilers (60%). In crows, of 16 isolates, 93.8% were C. jejuni and 6.2% were C. coli. Definitive identification of C. jejuni by PCR revealed positive results in 74.1% of 243 analysed isolates. Findings in this study indicate high prevalence of thermophilic campylobacters in humans, chickens and crows in Morogoro, and a higher infection rate of C. jejuni than that of C. coli in different animal species. Age of humans and location of chickens were identified as risk factors for thermophilic Campylobacter infections. Positive isolates to biochemical tests that indicated negative results on PCR indicates the additional value of PCR for definitive diagnosis of C. jejuni.
Rabies, an Emerging Disease in Korea
Authors: Kim C.‐H. , Lee C.‐G. , Yoon H.‐C. , Nam H.‐M. , Park C.‐K. , Lee J.‐C. , Kang M.‐I. , Wee S.‐H.
Published: March 2006

Since the first rabies case was reported in a dog in 1907, the disease was enzootic up to 1975 in Korea. After a steady decrease in the number of rabies cases from 1976 to 1984, no case was reported for 8 years from 1985 to 1992. Then, a resurgence of the disease was noted in 1993, and a continuous increase of rabies cases was observed during the following years. This report provides information on rabies in South Korea during the reemerging period 1993–2003. A total of 364 rabies cases in five different animal species and five deaths in human beings as a result of rabies were reported. Cattle and dogs accounted for 46.4% and 40.4% of total animal cases, respectively, and raccoon dogs commanded an overwhelming majority (44/48) of rabies cases in wildlife animal species. All animal and human rabies cases occurred only in two provinces, Gyeonggi and Gangwon; majority of them in two counties of Gyeonggi and one county of Gangwon province that border the demilitarized zone. From the three counties, the disease continued to expand to the other areas of the two provinces. The average monthly frequency of animal rabies cases during the 11‐year period peaked in January, and the incidence was highest during winter. There were three major rabies outbreaks in animals and the number of animal rabies cases increased with time. Data indicate that the temporal patterns were attributable to the ethology of raccoon dogs in the areas of outbreak.
A Virological Survey in Migrating Waders and Other Waterfowl in One of the Most Important Resting Sites of Germany
Authors: Hlinak A. , Mühle R. U. , Werner O. , Globig A. , Starick E. , Schirrmeier H. , Hoffmann B. , Engelhardt A. , Hübner D. , Conraths F. J. , Wallschläger D. , Kruckenberg H. , Müller T.
Published: March 2006

Wild birds are considered a potential reservoir or a carrier of viral diseases and may therefore play a role in the epidemiology of economically important or zoonotic diseases. In 2001 and 2002, a survey with special emphasis on virus isolation in migrating waders and some other birds were conducted. In one of the most important inland resting sites for migratory waterfowl, tracheal and cloacal swabs were collected from 465 waders representing 19 different species, and 165 other birds that were not captured on purpose. A total of 42 avian viruses were isolated, 34 of these were identified as paramyxoviruses (PMVs). The majority of isolates came from waders and wild ducks, and were characterized as PMV‐1. In contrast, PMV‐4 was found in wild ducks only, PMV‐6 was mainly detected in wader species. Four avian influenza viruses (AIVs), belonging to H4 and H3 haemagglutinin subtype, were isolated from wild duck species. Furthermore, four reo‐like viruses were isolated from one particular wader species for the first time. The majority of virus positive birds were <1 year old and did not show any clinical symptoms. There was no evidence for the presence of West Nile virus in these birds. These results confirm that the restricted resting sites in Western Europe must be considered as important locations for the intra‐ and interspecies transmission of avian viruses.
Digestive tract ontogeny of Dicentrarchus labrax: Implication in osmoregulation
Authors: Giffard‐Mena Ivone , Charmantier Guy , Grousset Evelyse , Aujoulat Fabien , Castille René
Published: March 2006

The ontogeny of the digestive tract (DT) and of Na+/K+‐ATPase localization was investigated during the early postembryonic development (from yolk sac larva to juvenile) of the euryhaline teleost Dicentrarchus labrax reared at two salinities: seawater and diluted seawater. Histology, electron microscopy and immunocytochemistry were used to determine the presence and differentiation of ion transporting cells. At hatching, the DT is an undifferentiated straight tube over the yolk sac. At the mouth opening (day 5), it comprises six segments: buccopharynx, esophagus, stomach, anterior intestine, posterior intestine and rectum, well differentiated at the juvenile stage (day 72). The enterocytes displayed ultrastructural features similar to those of mitochondria‐rich cells known to be involved in active ion transport. At hatching, ion transporting cells lining the intestine and the rectum exhibited a Na+/K+‐ATPase activity which increased mainly after the larva/juvenile (20 mm) metamorphic transition. The immunofluorescence intensity was dependent upon the stage of development of the gut as well as on the histological configuration of the analyzed segment. The appearance and distribution of enteric ionocytes and the implication of the DT in osmoregulation are discussed.
Rap‐GEF/Rap signaling restricts the formation of supernumerary spermathecae in Drosophila melanogaster
Authors: Singh Shree R. , Oh Su‐Wan , Liu Wei , Chen Xiu , Zheng Zhiyu , Hou Steven X.
Published: March 2006

Sperm storage in the female is a key factor for reproductive success in a variety of organisms, including Drosophila melanogaster. The spermathecae (SP) are the Drosophila organs for long‐term storage. While wild‐type female flies have two SP, occasionally, three or four SP have been observed in mutant flies. However, the molecular mechanism of SP formation is unknown. Here we show that loss of function of a Drosophila Rap‐GEF (GEF26) result in an occurrence of the supernumerary SP; females have three SP (varies from 11 to 62% in different allele combinations) instead of the normal two SP. In addition, the Gef26 mutant flies also have ectopic wing veins and extra mechanosensory organs. The supernumerary SP phenotype of the Gef26 mutation can be enhanced by the Drosophila Rap mutations and rescued by overexpressing the cell adhesion molecule DE‐cadherin. These data suggest that the Rap‐GEF/Rap signaling controls the formation of supernumerary spermathecae through modulating cell adhesion in Drosophila.
Peroxisome proliferator‐activated receptor γ ligands stimulate myeloid differentiation and lipogenensis in human leukemia NB4 cells
Authors: Yasugi Etsuko , Horiuchi Akiko , Uemura Isao , Okuma Emiko , Nakatsu Masami , Saeki Kumiko , Kamisaka Yasushi , Kagechika Hiroyuki , Yasuda Kazuki , Yuo Akira
Published: March 2006

Peroxisome proliferator‐activated receptor γ (PPARγ) plays a central role in adipocyte and macrophage differentiation. Pioglitazone (Actos, AD4833), an antidiabetic drug, and 15‐deoxy‐Δ12,14‐prostaglandin J2 (PGJ2) have recently been identified as synthetic and natural ligands for PPARγ, respectively. In this study, we examined the effects of PPARγ ligands on differentiation and lipogenesis in promyelocytic leukemia NB4 cells, in which PPARγ protein was expressed and ligand‐stimulated PPARγ‐specific transcription of adipocyte fatty‐acid binding protein was confirmed. Treatment with PPARγ ligand (AD4833 or PGJ2) alone markedly suppressed proliferation but did not induce differentiation. The combined treatment of the cells with PPARγ ligand and all‐trans retinoic acid (ATRA) synergistically induced myelocytic differentiation, as determined by nitroblue tetrazolium reducing ability and cell morphology. During these processes of differentiation, we observed marked accumulation of lipid droplets in the cytoplasm. The cellular triacylglycerol levels increased 2.7‐fold after treatment with the inducers. Simultaneously, BODIPY‐fatty acid was incorporated into the cytosol and concentrated in lipid droplets. The biosynthesis of triacylglycerol‐containing BODIPY‐fatty acids was increased twofold in differentiated cells. These findings clearly demonstrate that treatment with PPARγ ligands not only induced differentiation but also stimulated lipogenesis in NB4 cells, indicating a close association between differentiation and lipogenesis in PPARγ‐stimulated human myeloid cells.
Programmed cell death of follicular epithelium during the late developmental stages of oogenesis in the fruit flies Bactrocera oleae and Ceratitis capitata (Diptera, Tephritidae) is mediated by autophagy
Authors: Nezis Ioannis P. , Stravopodis Dimitrios J. , Margaritis Lukas H. , Papassideri Issidora S.
Published: March 2006

In the present study, we describe the features of programmed cell death of ovarian follicle cells, occurring during the late developmental stages of oogenesis in the olive fruit fly, Bactrocera oleae and the medfly, Ceratitis capitata. During stage 14, the follicle cells contain autophagic vacuoles, and they do not exhibit caspase activity in all parts of the egg chamber. Their nuclei are characterized by condensed chromatin, accompanied with high‐ but not low‐molecular weight DNA fragmentation events exclusively detected in distinct cells of the anterior pole. These data argue for the presence of an autophagy‐mediated cell death program in the ovarian follicle cell layer in both species. The above results are likely associated with the abundant phagocytosis observed at the entry of the lateral oviducts, where numerous cell bodies are massively engulfed by epithelial cells. We strongly believe that during the termination of the above Dipteran oogenesis, an efficient mechanism of absorption of the degenerated follicle cells is selectively activated, in order to prevent the blockage of the ovarioles and thus robustly support the physiological completion of the ovulation process.
Small GTPase RacF2 affects sexual cell fusion and asexual development in Dictyostelium discoideum through the regulation of cell adhesion
Authors: Muramoto Tetsuya , Urushihara Hideko
Published: March 2006

Cells of Dictyostelium discoideum become sexually mature when submerged and in darkness, and fuse with opposite mating‐type cells as gametes. The gene for a Rho GTPase, RacF2, is one of the extremely gamete‐enriched genes (>100‐fold) identified by us previously. Here, we isolated knockout, overexpression, constitutively active and dominant negative mutants of RacF2, and analyzed their phenotypes. These mutants showed anomalies in the extent of sexual cell fusion and asexual development as well as in EDTA‐sensitive cell–cell adhesion. It is suggested that RacF2 controls the process of sexual and asexual development through the regulation of cellular adhesiveness. An analysis of the expression of all 18 rac family genes by real‐time polymerase chain reaction revealed that four additional genes, rac1b, rac1c, racF1 and racG, were induced during maturation, suggesting their possible involvement in sexual cell interactions.