bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD177 polymorphisms: correlation between high‐frequency single nucleotide polymorphisms and neutrophil surface protein expression
Authors: Caruccio Lorraine , Walkovich Kelly , Bettinotti Maria , Schuller Randy , Stroncek David 
Published: December 2004

BACKGROUND:  Human neutrophil antigen‐2a (NB1) is located on NB1 glycoprotein, which is expressed on subpopulations of neutrophils. PRV‐1 is a gene that is over‐expressed in neutrophils from patients with polycythemia rubra vera. The gene encoding NB1 differs from PRV‐1 at four nucleotides. The purpose of this study was to determine if PRV‐1 and NB1 are alleles of the same gene or two separate genes; and, moreover, if they are alleles, to explore potential correlations to NB1 glycoprotein expression.STUDY DESIGN AND METHODS:  Primer pairs were used to amplify WBC DNA in the regions surrounding the four NB1 polymorphic sites. The four resulting amplicons were sequenced. The size of the neutrophil population in each donor that stained brightly with CD177 antibody was assessed by flow cytometry.RESULTS:  If PRV‐1 and NB1 are separate genes, then all people should be heterozygous for the PRV‐1/NB1 polymorphisms. Because 11 of 23 donors were homozygous for PRV‐1 polymorphisms at all four sites, PRV‐1 and NB1 are alleles of the same gene, CD177. When the sequenced exons were compared with PRV‐1, 13 single nucleotide polymorphisms (SNPs) that result in amino acid changes were found. The G42C NB1 polymorphism, found in 10 donors, was the most common SNP. The size of the CD177 bright‐staining neutrophil population was greater in 42C/C donors than in 42G/G donors (66 ± 20% vs. 41 ± 21%, p = 0.004).CONCLUSIONS:  PRV‐1 and NB1 are alleles of the polymorphic gene CD177. The most common SNP in bp 42 predicted an amino acid change that may have an effect on protein expression.
T‐Cell Activation and the Development of an Apoptosis‐Resistant CD45RO+ T‐Cell Population
Authors: Müller M. , Grunewald J. , Gigliotti D. , Eklund A. , Stridh H. 
Published: December 2004

AbstractGrowing experimental evidence supports a broadening role for the caspases; not only do they participate in the process of apoptosis but also in the control of the cell cycle and cellular proliferation. The biological role of the caspases in the process of T‐cell activation and proliferation is still not defined. In the present study, we propose a potential role, by demonstrating an association of T‐cell receptor‐mediated caspase activity with the development of an apoptosis‐resistant memory CD45RO+ T‐cell population. As previously shown by us, a time‐dependent induction of caspase activity, in the absence of apoptosis, can be observed in CD3‐stimulated human peripheral blood lymphocytes. We here show that a population of CD45RO+ cells, with activated caspase‐3 and with resistance to tributyltin‐induced apoptosis, develops after 3 days of stimulation. A concomitant expression of the anti‐apoptotic protein Bcl‐xL accompanied the caspase activity and the development of the apoptosis‐resistant phenotype. Finally, upon co‐culturing with dexamethasone (DEX), the CD3‐induced caspase‐3 activity was blocked. During this condition, the expression of the activation marker HLA‐DR as well as the cellular proliferative response was strongly suppressed. The development of memory cells with a CD45RO+ phenotype was also blocked. Our data support the hypothesis that caspase‐3 activity, observed in CD3‐stimulated cells, may be an important component in the proliferation process and, furthermore, might play a role for the development of memory T cells, and DEX inhibits this process.
Immunoglobulin G3 and Immunoglobulin M Isotype Plasma Levels are Influenced by Interleukin‐1α Genotype
Authors: Kilpinen S. , Laine S. , Hulkkonen J. , Hurme M. 
Published: December 2004

AbstractThe immunoglobulin (Ig) plasma levels are known to be, at least partially, genetically regulated, but all the genes involved are not known. Interleukin‐1 (IL‐1) is a potent proinflammatory cytokine able to serve as an adjuvant for immune responses. IL‐1α gene is polymorphic, and at least one of the polymorphisms has been identified in the 5′ regulatory region of the promoter, a biallelic base exchange (C→T) at position −889.We set out to study whether the IL‐1α genotype might contribute to the genetic component seen in the steady‐state antibody levels of healthy individuals.Four hundred healthy blood donors (218 males and 182 females) were genotyped, and the plasma levels of IgM, IgG as well as IgG subclasses were measured.An association was found between IgG3 plasma levels and the IL‐1α genotype; the 1.1 homozygotes had increased IgG3 levels compared with the 1.2 heterozygotes (P < 0.001 in males and P = 0.04 in females, Mann–Whitney U‐test). A similar significant association was also found between IgM plasma levels and the IL‐1α genotype in males, but it was no longer present in females; the 1.1 homozygotes had higher IgM levels than the 2.2 homozygotes (P = 0.03, Mann–Whitney U‐test).The data suggest that IL‐1α‐mediated signals are critical for IgG3 and IgM responses, which are induced by thymus‐independent antigens and are important in activating complement.
Brown Norway Rat Ovalbumin‐Specific Immunoglobulin E Antibodies Increase the Human Basophil Expression of CD63 Marker
Authors: Bellou A. , Saint‐Laudy J. , Knippels L. , Montémont C. , Vauthier E. , Gerard P. , Pellegrom H. , Koerkamp E. K. , Lesesve J. F. , Guéant J. L. , Lambert H. , Mallié J. P. 
Published: December 2004

AbstractAnaphylactic shock is an immunoglobulin E (IgE)‐dependent hypersensitivity. Biological tests like leucocyte histamine release (LHR) and human basophil activation (HBA), frequently used in human allergy, reflect both the amount of IgE fixed on cells and the cellular reactivity. To assess whether serum‐specific IgE from Brown Norway (BN) rats prepared for ovalbumin (OVA)‐induced anaphylactic shocks can activate human basophils which has a potential interest in experimental allergy: such a test could rapidly assert an IgE sensitization in laboratory animals genetically T‐helper 2 (Th2)‐predisposed. Rats (n = 39) were immunized three times (day 0, day 5 and day 21) with OVA injected subcutaneously. One week after the third immunization, a shock was induced with an intravenous (i.v.) bolus of OVA. Sensitization was assessed by passive cutaneous anaphylaxis (PCA) test and dosages of serum IgE antibodies anti‐OVA by enzyme‐linked immunosorbent assay. Blood basophils were counted before and during the shock. Before the shock induction (at day 21), an LHR test was performed on rat blood, and human basophils were sensitized with rat sera. HBA was demonstrated by the increase in the percentage of cells expressing CD63 antigen membrane, measured by flow cytometry. Twenty‐one days after the first subcutaneous (s.c.) immunization, the rat serum induced a significant HBA. HBA was observed neither with the same serum previously heated nor with the serum from nonimmunized rats (NIRs). OVA‐specific IgEs were significantly increased in immunized rat (IR) serum. The PCA test was negative when the serum was previously heated (56 °C). We never observed any circulating basophils, and LHR test was negative. After OVA i.v. administration, all IRs died rapidly. HBA testing strongly suggests a mediation by specific IgE in the increase of CD63 in BN rats. Thus, HBA test seems useful in assessing whether an experimental allergy was induced in animals genetically predisposed to an immune response, Th2‐mediated, like BN rat. We also conclude that rat basophil activation does not participate in the histamine release during anaphylactic shock in sensitized BN rats.
Long‐Term CD4+ and CD8+ Memory T Cells Developed in Severe Combined Immunodeficiency Mice during Homoeostasis Exhibit Differences in Sensitivity to Antigen
Authors: Pettersson F. Ekholm , Grönvik K.‐O. 
Published: December 2004

AbstractT cells transferred in small numbers to lymphopenic hosts proliferate spontaneously, and naïve T cells turn into memory cells without complete cellular reconstitution of the lymphoid compartment. In this study, neonatal severe combined immunodeficiency mice were treated with peripheral CD4+ or CD8+ T cells purified from the spleen of syngeneic C.B‐17 mice. At 2 weeks and more pronounced at 10 weeks post treatment, a majority of the residing donor T cells showed memory phenotype, with high expression of CD44 and an early onset of proliferation and cytokine production upon stimulation. These memory type of donor cells were sustained in numbers for at least 1.5 years post treatment in a homoeostatic fashion, recognized by normal CD4/CD8 ratio and no bias towards type 1 or type 2 immune response. Furthermore, amongst the memory type of cells, there was a striking difference in their response, where the CD8+ donor cells had higher threshold for stimulation than the CD4+ donor cells.
Nodular celestite in the Chihsia Formation (Middle Permian) of south China
Authors: Yan Jiaxin , Carlson Ernest H. 
Published: December 2004

ABSTRACTThe middle Permian Chihsia Formation of south China accumulated on a shallow shelf, and consists mainly of black to dark grey micritic limestone rich in chert nodules and organic matter. A unique type of nodular crystal cluster is distributed widely in the carbonate succession. Most crystal clusters consist of calcite. Some, however, are composed of celestite, and geochemical, microscopic and crystal morphological data suggest that celestite was the precursor of the calcite. The celestite developed displacively within the sediments during early diagenesis, before compaction and before local dolomitization of the host rock. Similar strontium isotopic values were obtained from the celestite clusters, replacement calcite, vein calcite and host rock. The values are within the range of middle Permian sea water. The strontium in the celestite was furnished chiefly by either diagenetic alteration of strontium‐rich marine aragonite to strontium‐poor calcite, or aragonite dissolution induced by aerobic oxidation of organic matter, or both. The sulphur isotopic values of the celestite are about 6–11‰ heavier than the sulphur isotopic value of sulphate in coeval sea water. Based on geological context, this difference is attributed to microbial reduction of porewater sulphate in the Chihsia sediments.
Analysis of large DMA from soybean (Glycine max L. Merr.) by pulsed‐field gel electrophoresis
Authors: Honeycutt Rhonda J. , Sobral Bruno W.S. , McClelland Michael , Atherly Alan G. 
Published: December 2004

SummaryThe technique of pulsed‐field gel electrophoresis (PFE) has been used to study chromosomal regions and entire genomes of several organisms. Techniques are presented for the isolation of high molecular weight DMA from embedded soybean protoplasts and the conditions for separating large DMA fragments using PFE. Digestion was detected by Southern hybridization using single copy nodulin clones. These data are being used to generate a physical map of the nodulin region(s) of the soybean genome.
Correlated clustering and virtual display of gene expression patterns in the wheat life cycle by large‐scale statistical analyses of expressed sequence tags
Authors: Ogihara Yasunari , Mochida Keiichi , Nemoto Yasue , Murai Koji , Yamazaki Yukiko , Shin‐I Tadasu , Kohara Yuji 
Published: December 2004

SummaryCompared to rice, wheat exhibits characteristic growth habits and contains complex genome constituents. To assess global changes in gene expression patterns in the wheat life cycle, we conducted large‐scale analysis of expressed sequence tags (ESTs) in common wheat. Ten wheat tissues were used to construct cDNA libraries: crown and root from 14‐day‐old seedlings; spikelet from early and late flowering stages; spike at the booting stage, heading date and flowering date; pistil at the heading date; and seeds at 10 and 30 days post‐anthesis. Several thousand colonies were randomly selected from each of these 10 cDNA libraries and sequenced from both 5′ and 3′ ends. Consequently, a total of 116 232 sequences were accumulated and classified into 25 971 contigs based on sequence homology. By computing abundantly expressed ESTs, correlated expression patterns of genes across the tissues were identified. Furthermore, relationships of gene expression profiles among the 10 wheat tissues were inferred from global gene expression patterns. Genes with similar functions were grouped with one another by clustering gene expression profiles. This technique might enable estimation of the functions of anonymous genes. Multidimensional analysis of EST data that is analogous to the microarray experiments may offer new approaches to functional genomics of plants.
In vitro analysis of transcription initiation and termination from the Lhcb1 gene family in Nicotiana sylvestris: detection of transcription termination sites
Authors: Hasegawa Keiko , Yukawa Yasusi , Sugiura Masahiro 
Published: December 2004

SummaryGenes encoding chlorophyll a/b‐binding proteins of photosystem II (Lhcb) constitute a multigene family. Nine Lhcb1 genes have previously been isolated from the tobacco species, Nicotiana sylvestris, and the transcription initiation sites in vivo have been mapped. Reaction conditions from a previously developed in vitro transcription system from tobacco cultured cells were optimized for the Lhcb1 genes. Transcription initiation sites in vitro predominantly coincided with those found in vivo and were typically cytidines, a system unique to N. sylvestris Lhcb1 genes. CTC*A (C* for initiation site) was a consensus motif for the initiation region in vitro, as reported in vivo. Mutation analysis defined functionally that the TATA box is essential for transcription initiation and that the CTCA motif is a determinant of transcription initiation sites but not transcript levels. Polyadenylation sites were determined from in vivo transcripts, located 12–21 nt downstream from likely poly(A) signals. Four major 3′‐ends of in vitro transcripts from Lhcb1*6 were detected, 40–300 nt downstream of the poly(A) site, suggestive of multiple, discrete transcription termination sites. These 3′‐ends are mapped in or nearby T‐rich sequences.
Increased dehydrin promoter activity caused by HvSPY is independent of the ABA response pathway
Authors: Robertson Masumi 
Published: December 2004

SummaryA barley SPINDLY protein, HvSPY, is a negative regulator of gibberellin (GA) action. It is also found to be a positive regulator of the promoter of a barley dehydrin (Dhn) gene which is abscisic acid (ABA) upregulated. To investigate whether HvSPY acts through the ABA signaling pathway to upregulate the Dhn promoter, functional characterization was carried out by co‐bombardment experiments. These experiments used Dhn promoter‐GUS reporter constructs and an effector construct to overexpress HvSPY protein in barley aleurone. ABA dose–response experiments with and without HvSPY overexpression showed that the induction by HvSPY occurred in addition to the ABA effect. Gibberellic acid (GA3) did not reduce the induction by ABA, but it had a small, although significant, effect on the ability of HvSPY to upregulate. The induction of promoter activity of Dhn by HvSPY required the intact protein, and a small deletion in the tetratricopeptide repeat (TPR) region reduced this ability significantly. When a promoter region containing an element for ABA responsiveness was mutagenized or deleted, the mutant promoters lost ABA responsiveness but remained responsive to HvSPY. In addition, HvSPY did not increase promoter activities of other ABA‐upregulated genes. Taken together, these results indicate that HvSPY and ABA both regulate promoter activity of Dhn, and that HvSPY acts independently of the ABA signaling pathway.
Flower development in carrot CMS plants: mitochondria affect the expression of MADS box genes homologous to GLOBOSA and DEFICIENS
Authors: Linke Bettina , Nothnagel Thomas , Börner Thomas 
Published: December 2004

SummaryMaternally inherited defects in the formation of male flower organs leading to cytoplasmic male sterility (CMS) indicate an involvement of mitochondrial genes in the control of flower formation. In the ‘carpeloid’ CMS type of carrot, stamens are replaced by carpels. The florets thus resemble well‐investigated homeotic flower mutants of Arabidopsis and Antirrhinum, in which organ identity is impaired because of the mutation of specific nuclear MADS box genes. We have isolated five cDNAs encoding MADS box proteins (DcMADS1–5) from a flower‐specific library of carrot. Structural features deduced from their sequence and transcript patterns in unmodified carrot flowers determined by in situ hybridisation relate them to known MADS box transcription factors involved in specification of flower organs. In ‘carpeloid’ CMS flowers, we detected a distinctly reduced expression of DcMADS2 and DcMADS3, homologues of the Antirrhinum genes GLOBOSA and DEFICIENS. Our data strongly suggest that the ‘carpeloid’ CMS phenotype is caused by a cytoplasmic (mitochondrial) effect on the expression of two MADS box factors specifying organ development at whorls 2 and 3 of carrot flowers.
Automated whole mount localisation techniques for plant seedlings
Authors: Friml Jiří , Benková Eva , Mayer Ulrike , Palme Klaus , Muster Gerhard 
Published: December 2004

SummaryPlant biology is currently experiencing a growing demand for easy and reliable mRNA and protein localisation techniques. Here, we present novel whole mount in situ hybridisation and immunolocalisation protocols, suitable to localise mRNAs and proteins in Arabidopsis seedlings. We demonstrate that these methods can be used in different organs of Arabidopsis seedlings as well as in other plant species. In order to achieve better reproducibility and higher throughput, we modified these protocols for automation to be performed by a liquid handling robot. In addition, we show that other procedures such as reporter enzyme assays and tissue clearing can be similarly automated. We present examples of application of our protocols including mRNA localisation and proteins and epitope tag (co)localisations which demonstrate that these methods provide reliable and versatile tools for expression, localisation and anatomical studies in plants.
The ABSCISIC ACID INSENSITIVE 3 (ABI3) gene is modulated by farnesylation and is involved in auxin signaling and lateral root development in Arabidopsis
Authors: Brady Siobhan Mary , Sarkar Sara F. , Bonetta Dario , McCourt Peter 
Published: December 2004

SummaryGenetic screens have identified a number of genes that regulate abscisic acid (ABA) responsiveness in Arabidopsis. Using a combination of suppressor screens and double mutant analysis, we have determined a genetic relationship for a number of these ABA response loci. Based on germination in the presence of exogenous ABA, the ABI1 and ABI2 phosphatases act at or upstream of the ERA1 farnesyl transferase and the ABI3 and ABI5 transcription factors act at or downstream of ERA1. In contrast with ABI3 and ABI5, the ABI4 transcription factor appears to act at or upstream of ERA1. Based on reporter gene constructs, the upstream regulation of ABI3 by ERA1 occurs at least partially at the level of transcription, suggesting that this lipid modification is required to attenuate ABI3 expression. Similar experiments also indicate that ABI3 is auxin inducible in lateral root primordia. Related to this, loss‐of‐function abi3 alleles show reduced lateral root responsiveness in the presence of auxin and an auxin transport inhibitor, and era1 mutants have increased numbers of lateral roots. These results suggest the possibility that genes identified through ABA responsive germination screens such as ERA1 and ABI3 have functions in auxin action in Arabidopsis.
High‐Cot sequence analysis of the maize genome
Authors: Yuan Yinan , SanMiguel Phillip J. , Bennetzen Jeffrey L. 
Published: December 2004

SummaryHigher eukaryotic genomes, including those from plants, contain large amounts of repetitive DNA that complicate genome analysis. We have developed a technique based on DNA renaturation which normalizes repetitive DNA, and thereby allows a more efficient outcome for full genome shotgun sequencing. The data indicate that sequencing the unrenatured outcome of a Cot experiment, otherwise known as High‐Cot DNA, enriches genic sequences by more than fourfold in maize, from 5% for a random library to more than 20% for a High‐Cot library. Using this approach, we predict that gene discovery would be greater than 95% and that the number of sequencing runs required to sequence the full gene space in maize would be at least fourfold lower than that required for full‐genome shotgun sequencing.
Transfusion for anaemia in the district hospital setting
Authors: Ridley J. , Revell P. , Ainsworth M. , Pryce A. 
Published: December 2004

summary. We examined the practice of transfusion for anaemia in our hospitals using an audit method, with standards derived from guidelines published by the British Committee for Standards in Haematology.Documentation was generally good, but simple tests were not performed prior to transfusion in many cases when the diagnosis was not known.
Early primary immune response against erythrocytes: a case report
Authors: Albiero A. L. , Novaretti M. C. Z. , Llacer P. E. D. , Chamone D. A. F. 
Published: December 2004

summary. Background Primary immune response against red blood cell (RBC) antigens often takes weeks or months to be detected. In previous reports, for children receiving multiple units of blood components, ranging from five to 81 units, the elapsed time between the first RBC transfusion and antibody detection ranged from 18 to 78 days. Cytomegalovirus (CMV) is sometimes associated with immunohaematologic findings and may modulate immune response.Case report A 24‐week‐old male infant with interstitial pneumonia and hepatitis because of CMV developed an RBC auto antibody and two RBC alloantibodies: anti‐Jka, detected in tube 11 days after a single RBC transfusion, and anti‐K, detected only in papain gel test 18 days later.Conclusion As anti‐Jka is not a naturally occurring antibody, this is the most rapid primary immune response against an RBC antigen after a single RBC transfusion ever described, in the youngest child ever described.
Factors influencing the care‐seeking behaviour of chest symptomatics: a community‐based study involving rural and urban population in Tamil Nadu, South India
Authors: Sudha G. , Nirupa C. , Rajasakthivel M. , Sivasusbramanian S. , Sundaram V. , Bhatt S. , Subramaniam K. , Thiruvalluvan E. , Mathew R. , Renu G. , Santha T. 
Published: December 2004

SummaryOur aim was to identify the factors that influence the care‐seeking behaviour of chest symptomatics in urban and rural areas in South India. We conducted in‐depth interviews with 649 participants: 80% of 310 urban residents and 63% of 339 rural people had sought care (P < 0.01), 93% within 1 month of onset of symptoms. Private health care facilities were the first and preferred point of contact for 57% of urban and 48% of rural participants; the major reasons were proximity to residence and their perception that good‐quality care would be available there. Symptomatics who did not seek care attributed their inaction to insufficient severity of symptoms (51%), unaffordability (46%) and lack of time due to work pressures (25%). Socio‐economic factors such as literacy and family income significantly influenced care‐seeking behaviour. Our results indicate that most chest symptomatics seek care promptly; their initial response is to go to the nearest private health care facility, shifting to another if they are dissatisfied. Fifty per cent of the participants who did not seek care felt that their symptoms were not severe.