bluuebunny/crossref_metadata_2025 huggingface dataset:
Genotype–phenotype correlation in MYH9‐related thrombocytopenia
Authors: Dong Fan , Li Sufeng , Pujol‐Moix Núria , Luban Naomi L. C. , Shin Sang Won , Seo Jae Hong , Ruiz‐Saez Arlette , Demeter Judit , Langdon Scott , Kelley Michael J.
Published: July 2005

Mutation of the non‐muscle myosin heavy chain type II‐A results in MYH9‐related hereditary macrothrombocytopenia (HMTC), including four autosomal dominant platelet disorders: May‐Hegglin anomaly (MHA), Sebastian (SBS), Fechtner (FS) and Epstein (EPS) syndrome. Denaturing high‐performance liquid chromatography (DHPLC) was optimised for rapid screening of the seven exons harbouring all but one of the previously reported mutations of MYH9. Individuals from 13 families with phenotypes suggestive of MYH9‐related HMTC were screened for mutations by DHPLC followed by direct sequencing of samples with aberrant column retention time. Mutations were identified in all 13 families. Six distinct missense heterozygous mutations were found in 10 families, including six families with MHA or SBS (E1841K, D1424N), three families with FS (R702H, R1165C, and D1424Y), and one family with EPS (S96L). A truncating mutation (R1933X) was found in three MHA families. A review of all published mutations suggests that mutation in the C‐terminal coiled coil region or truncation of the tailpiece is associated with haematological‐only phenotype, while mutation of the head ATPase domain frequently is associated with nephropathy and/or hearing loss. Mutations of other regions have intermediate expression of non‐haematological characteristics. Further study is required to confirm these associations and understand the molecular basis for this genotype–phenotype relationship.
The prognostic value of CD38 expression in chronic lymphocytic leukaemia patients studied prospectively at diagnosis: a single institute experience
Authors: Gentile Massimo , Mauro Francesca Romana , Calabrese Elisabetta , Propris Maria Stefania De , Giammartini Elena , Mancini Francesca , Milani Maria Laura , Guarini Anna , Foà Robin
Published: July 2005

The purpose of this study was to assess in chronic lymphocytic leukaemia (CLL) patients the prevalence and clinical impact of CD38 expression, evaluated prospectively at disease presentation, and to verify whether this parameter changes over time. In 242 consecutive and untreated CLL patients, the percentage of CD38+cases, according to the 7%, 20% and 30% cut‐off points, was 21%, 17% and 14%, respectively. Using the 7% threshold, CD38 positivity correlated with male sex, intermediate and high‐risk (Rai I–IV) disease, lower Hb and platelet levels, and higher lymphocyte count. Furthermore, patients with a CD38 expression ≥7% showed a significantly lower 3‐year probability of treatment‐free survival (TFS) than CD38−patients (P < 0·0001). At multivariate analysis, CD38 expression remained significantly associated to TFS, together with stage, lymphocyte count and morphology. Also, in the 146 patients with stage 0 CLL a CD38 expression ≥7% identified a subgroup of patients with a significantly lower 3‐year probability of TFS (P = 0·0005). Furthermore, this parameter did not change in 30 of 31 (97%) re‐evaluated patients. In conclusion, this study indicates that, when tested at diagnosis and on fresh material, a CD38 expression ≥7% is an important parameter for the identification of early CLL patients with more aggressive disease and that its expression remains stable over time.
Iron acquisition mechanisms of Flavobacterium psychrophilum
Authors: Møller J D , Ellis A E , Barnes A C , Dalsgaard I
Published: July 2005

Forty strains of Flavobacterium psychrophilum were tested for the production of siderophores using the universal Chrome Azurol S (CAS) assay. The majority of the strains (85%) were CAS positive (CAS+) and some (15%) were CAS negative (CAS−). The cryptic plasmid pCP1 was carried by all positive strains and was lacking from negative strains. While a weak catechol reaction was detectable in CAS+ culture supernatants, the CAS reaction was, to some extent, heat sensitive, questioning whether the positive reaction was caused only by siderophores. The ability to grow in vitro under iron‐restricted conditions did not correlate with the CAS reactivity, as growth of both CAS+ and CAS− strains was similarly impaired under iron restriction induced by 2,2 dipyridyl. Suppressed growth under these conditions was restored by addition of FeCl3, haemoglobin and transferrin for both CAS+ and CAS− strains.
Potential use of a Yersinia ruckeri O1 auxotrophic aroA mutant as a live attenuated vaccine
Authors: Temprano A , Riaño J , Yugueros J , González P , de Castro L , Villena A , Luengo J M , Naharro G
Published: July 2005

The aroA gene of Yersinia ruckeri, which encodes 5‐enolpyruvylshikimate 3‐phosphate synthase, was insertionally inactivated with a DNA fragment containing a kanamycin resistance determinant and reintroduced by allelic exchange into the chromosome of Y. ruckeri 21102 O1 by means of the suicide vector pIVET8. The Y. ruckeri aroA::Kanr mutant was highly attenuated when inoculated intraperitoneally into rainbow trout, with a 50% lethal dose of >5 × 107 CFU. The mutants were not recoverable from the internal organs 48 h post‐inoculation or later. The vaccination of rainbow trout with the AroA mutant as a live vaccine conferred significant protection (relative percentage survival = 90%) against the pathogenic wild‐type strain of Y. ruckeri.
European eels, Anguilla anguilla (L.), infected with Anguillicola crassus exhibit a more pronounced stress response to severe hypoxia than uninfected eels
Authors: Gollock M J , Kennedy C R , Brown J A
Published: July 2005

The parasite, Anguillicola crassus is a non‐native species that infects naive European eels, Anguilla anguilla, and causes pathological damage to the swimbladder, potentially compromising their ability to cope with hypoxic conditions. This study aimed to elucidate whether anguillicolosis exacerbates the stress responses to exposure to hypoxic water, conditions that have been implicated in mass mortalities of wild infected European eels. Blood parameters in infected and uninfected eels were measured during exposure to severe hypoxia over an 8‐h period. Infected fish showed significantly higher levels of plasma cortisol compared with uninfected eels after 4 h of hypoxia. Uninfected fish showed an almost twofold increase in plasma glucose after 8‐h exposure to hypoxia but infected fish showed no significant change, so that the plasma glucose concentration was significantly higher in uninfected eels than in infected eels. Both groups showed similar elevations in blood haematocrit, suggesting a similar catecholamine response in infected and uninfected eels. The lack of a hyperglycaemic response in infected eels, despite indirect evidence of a catecholamine response to hypoxia, may reflect an increase in glucose turnover. The data suggest that anguillicolosis results in a significantly greater corticosteroid stress response to hypoxia accompanied by a higher metabolic cost.
Different proteins bind to the butyrolactone receptor protein ARE sequence located upstream of the regulatory ccaR gene of Streptomyces clavuligerus
Authors: Santamarta Irene , Pèrez‐Redondo Rosario , Lorenzana Luis M. , Martìn Juan F. , Liras Paloma
Published: July 2005

Cell‐free extracts from Streptomyces clavuligerus, purified by elution from heparin‐agarose with an ARE‐containing DNA fragment or by salt elution chromatography, bind to a 26 nt ARE sequence, for butyrolactone receptor proteins (AREccaR). This sequence is located upstream of the ccaR gene, encoding the activator protein CcaR required for clavulanic acid and cephamycin C biosynthesis. The binding is specific for the ARE sequence as shown by competition with a 34 nt unlabelled probe identical to the ARE sequence. A brp gene, encoding a butyrolactone receptor protein, was cloned from S. clavuligerus. Sixty‐one nucleotides upstream of brp another ARE sequence (AREbrp) was found, suggesting that Brp autoregulates its expression. Pure recombinant rBrp protein binds specifically to the ARE sequences present upstream of ccaR and brp. A brp‐deleted mutant, S. clavuligerus Δbrp::neo1, produced 150–300% clavulanic acid and 120–220% cephamycin C as compared with the parental strain, suggesting that Brp exerts a repressor role in antibiotic biosynthesis. EMSA assays using affinity chromatography extracts from the deletion mutant S. clavuligerus Δbrp::neo1 lacked a high‐mobility band‐shift due to Brp but still showed a slow‐mobility band‐shift observed in the wild‐type strain. These results indicate that two different proteins bind specifically to the ARE sequence and modulate clavulanic acid and cephamycin C biosynthesis by its action on ccaR gene expression.
Weapons of mass retraction
Authors: Burrows Lori L.
Published: July 2005

Twitching motility is a unique form of bacterial propulsion on solid surfaces associated with cycles of extension, tethering and retraction of type IV pili (T4P). Although investigations over the last two decades in a number of species have identified the majority of the genes involved in this process, we are still learning how these pili are assembled and the mechanics by which bacteria use T4P to drag themselves from one place to another. Among the puzzles that remain to be solved is the mechanism by which hydrolysis of ATP is coupled to pilus assembly and disassembly, and how the cell envelope structure is modified to accommodate the passage of the pilus through the periplasm. Unravelling of these and other enigmas in the T4P system will not only teach us more about these important colonization and virulence factors, but also help us to understand related processes such as type II secretion, which relies on a set of proteins homologous to those in the T4P system, and bacterial conjugation, involving retractable pili belonging to the F‐like subgroup of the type IV secretion family. This review focuses on recent discoveries relating to the assembly and function of T4P in generation of twitching motility.
LuxS controls bacteriocin production in Streptococcus mutans through a novel regulatory component
Authors: Merritt Justin , Kreth Jens , Shi Wenyuan , Qi Fengxia
Published: July 2005

The oral pathogen Streptococcus mutans employs a variety of mechanisms to maintain a competitive advantage over many other oral bacteria which occupy the same ecological niche. Production of the bacteriocin, mutacin I, is one such mechanism. However, little is known about the regulatory mechanisms associated with mutacin I production. Previous work has demonstrated that the production of mutacin I greatly increased with cell density. In this study, we found that high cell density also triggered high level mutacin I gene transcription. However, this response was abolished upon deletion of luxS. Further analysis using real‐time reverse transcription polymerase chain reaction (RT‐PCR) demonstrated that in the luxS mutant transcription of both the mutacin I structural gene mutA and the mutacin I transcriptional activator mutR was impaired. Through microarray analysis, a putative transcription repressor annotated as Smu1274 in the Los Alamos National Laboratory Oral Pathogens Sequence Database was identified, which was strongly induced in the luxS mutant. Characterization of Smu1274, which we referred to as irvA, suggested that it may act as an inducible repressor to suppress mutacin I gene expression. A luxS and irvA double mutant regained the ability to produce mutacin I; whereas a constitutive irvA‐producing strain was impaired in mutacin I production. These findings reveal a novel regulatory pathway for mutacin I gene expression, which may provide clues to the regulatory mechanisms of other cellular functions regulated by luxS in S. mutans.
Defining the genetic differences between wild and domestic strains of Bacillus subtilis that affect poly‐γ‐dl‐glutamic acid production and biofilm formation
Authors: Stanley Nicola R. , Lazazzera Beth A.
Published: July 2005

Biofilms are communities of microbial cells that are encased in a self‐produced, polymeric matrix and  are  adherent  to  a  surface.  For  several  species of bacteria, an enhanced ability to form biofilms has been linked with an increased capability to produce exopolymers. To identify exopolymers of Bacillus subtilis that can contribute to biofilm formation, we transferred the genetic determinants that control exopolymer production from a wild, exopolymer‐positive strain to a domesticated, exopolymer‐negative strain. Mapping these genetic determinants led to the identification of γ‐poly‐dl‐glutamic acid (γ‐PGA) as an exopolymer that increases biofilm formation, possibly through enhancing cell–surface interactions. Production of γ‐PGA by Bacillus subtilis was known to be dependent on the two‐component regulator ComPA; this study highlighted the additional dependence on the DegS‐DegU, DegQ and SwrA regulator proteins. The inability of the domestic strain of B. subtilis to produce γ‐PGA was mapped to two base pairs; a single base pair change in the promoter region of degQ and a single base pair insertion in the coding region of swrA. Introduction of alleles of degQ and swrA from the wild strain into the domestic strain was sufficient to allow γ‐PGA production. In addition to controlling γ‐PGA production, ComPA and DegSU were also shown to activate biofilm formation through an as yet undefined pathway. The identification of these regulators as affecting γ‐PGA production and biofilm formation suggests that these processes are regulated by osmolarity, high cell density and phase variation.
Multiple drug resistance genes in malaria – from epistasis to epidemiology
Authors: Duraisingh Manoj T. , Refour Philippe
Published: July 2005

A decline in our ability to successfully treat patients with malaria infections of the parasitic protozoan Plasmodium falciparum with cheap quinoline drugs has led to a huge escalation in morbidity and mortality in recent years. Many approaches have been taken, including classical genetics, reverse genetics and molecular epidemiology, to identify the molecular determinants underlying this resistance. The contribution of the P. falciparum multidrug resistance gene, pfmdr1, to antimalarial resistance has been a source of controversy for over a decade since it was first identified. In the current issue of Molecular Microbiology, Sidhu and colleagues use powerful reverse genetics to demonstrate the importance of commonly occurring alleles of pfmdr1 in conferring resistance to the second‐line drugs quinine and sensitivity to the new alternatives mefloquine and artemisinin. They also elegantly highlight the importance of genetic background and epistasis between pfmdr1 and other potential modulators of drug resistance. Such molecular knowledge will facilitate surveillance/monitoring and aid the development of strategies for the reversal of resistance.
Frp1 is a Fusarium oxysporum F‐box protein required for pathogenicity on tomato
Authors: Duyvesteijn Roselinde G. E. , Van Wijk Ringo , Boer Yvonne , Rep Martijn , Cornelissen Ben J. C. , Haring Michel A.
Published: July 2005

Fusarium oxysporum f. sp. lycopersici is the causal agent of tomato wilt disease. In order to identify genes involved in its pathogenicity, we performed insertional mutagenesis. Mutant N40 had lost its pathogenicity completely, when tested in bioassays with tomato seedlings. Molecular characterization of mutant N40 revealed that the plasmid insertion had occurred in a gene that codes for a 60.2 kDa protein containing an F‐box motif. The gene was therefore designated as FRP1 (F‐box protein required for pathogenicity). Targeted FRP1 disruptants had lost their pathogenicity completely, and became fully virulent again upon re‐introduction of the FRP1 gene. This confirmed that the FRP1 gene is required for pathogenesis. In a yeast two‐hybrid assay Frp1 interacts with Skp1, suggesting involvement of an SCF ubiquitin ligase complex in pathogenicity. FRP1 is constitutively expressed during infection and under different culture conditions. Although growth, spore formation and germination on artificial media were not impaired, confocal laser scanning microscopy of a GFP‐marked mutant N40 and a GFP‐marked targeted FRP1 disruptant revealed that they were unable to colonize the roots.
A new mechanism for chloramphenicol, florfenicol and clindamycin resistance: methylation of 23S ribosomal RNA at A2503
Authors: Kehrenberg Corinna , Schwarz Stefan , Jacobsen Lene , Hansen Lykke H. , Vester Birte
Published: July 2005

The gene product of cfr from Staphylococcus sciuri confers resistance to chloramphenicol, florfenicol and clindamycin in Staphylococcus spp. and Escherichia coli. Cfr is not similar to any other known chloramphenicol resistance determinant. Comparative investigation of E. coli with and without a plasmid‐encoded Cfr showed a decreased drug binding to ribosomes in the presence of Cfr. As chloramphenicol/florfenicol and clindamycin have partly overlapping drug binding sites on the ribosome, the most likely explanation is that Cfr modifies the RNA in the drug binding site. This hypothesis was supported by drug footprinting data that showed both a decreased drug binding and an enhanced reverse transcriptase stop at position 2504, which corresponds to a modification at position A2503 at the drug binding site. A 45 n long RNA fragment containing the appropriate region was isolated and MALDI‐TOF mass spectrometry in combination with tandem mass spectrometry showed an additional methylation at position A2503. Moreover, reduced methylation was detected at nucleotide C2498. The results show that Cfr is an RNA methyltransferase that targets nucleotide A2503 and inhibits ribose methylation at nucleotide C2498, thereby causing resistance to chloramphenicol, florfenicol and clindamycin.
Protein phosphorylation is a prerequisite for intracellular Ca2+release and ion channel control by nitric oxide and abscisic acid in guard cells
Authors: Sokolovski Sergei , Hills Adrian , Gay Rob , Garcia‐Mata Carlos , Lamattina Lorenzo , Blatt Michael R.
Published: July 2005

Recent work has indicated that nitric oxide (NO) and its synthesis are important elements of signal cascades in plant–pathogen defence, and are a prerequisite for drought and abscisic acid (ABA) responses inArabidopsis thalianaandVicia fabaguard cells. NO regulates inward‐rectifying K+channels and Cl−channels ofViciaguard cells via intracellular Ca2+release. However, its integration with related signals, including the actions of serine–threonine protein kinases, is less well defined. We report here that the elevation of cytosolic‐free [Ca2+] ([Ca2+]i) mediated by NO in guard cells is reversibly inhibited by the broad‐range protein kinase antagonists staurosporine and K252A, but not by the tyrosine kinase antagonist genistein. The effects of kinase antagonism translate directly to a loss of NO‐sensitivity of the inward‐rectifying K+channels and background (Cl−channel) current, and to a parallel loss in sensitivity of the K+channels to ABA. These results demonstrate that NO‐dependent signals can be modulated through protein phosphorylation upstream of intracellular Ca2+release, and they implicate a target for protein kinase control in ABA signalling that feeds into NO‐dependent Ca2+release.
[Ala12]MCD peptide: a lead peptide to inhibitors of immunoglobulin E binding to mast cell receptors1
Authors: Buku A. , Condie B.A. , Price J.A. , Mezei M.
Published: July 2005

Abstract:  An effort was made to discover mast cell degranulating (MCD) peptide analogs that bind with high affinity to mast cell receptors without triggering secretion of histamine or other mediators of the allergic reaction initiated by immunoglobulin E (IgE) after mast cell activation. Such compounds could serve as inhibitors of IgE binding to mast cell receptors. An alanine scan of MCD peptide reported previously showed that the analog [Ala12]MCD was 120‐fold less potent in histamine‐releasing activity and fivefold more potent in binding affinity to mast cell receptors than the parent MCD peptide. Because this analog showed marginal intrinsic activity and good binding affinity it was subsequently tested in the present study as an IgE inhibitor. In contrast to MCD peptide, [Ala12]MCD showed a 50% inhibition of IgE binding to the FcεRIα mast cell receptor by using rat basophilic leukemia (RBL‐2H3) mast cells and fluorescence polarization. Furthermore, in a β‐hexosaminidase secretory assay, the peptide also showed a 50% inhibition of the secretion of this enzyme caused by IgE. An attempt was made to relate structural changes and biologic differences between the [Ala12]MCD analog and the parent MCD peptide. The present results show that [Ala12]MCD may provide a base for designing agents to prevent IgE/FcεRIα interactions and, consequently, allergic conditions.
Purification and mass spectroscopic analysis of human CB1 cannabinoid receptor functionally expressed using the baculovirus system
Authors: Xu W. , Filppula S.A. , Mercier R. , Yaddanapudi S. , Pavlopoulos S. , Cai J. , Pierce W.M. , Makriyannis A.
Published: July 2005

Abstract: The cannabinoid receptor 1 (CB1) cannabinoid receptor is an essential component of the cannabinergic system. It has been recognized as a therapeutic target for treating numerous diseases and is currently receiving considerable attention by the pharmaceutical community. Target‐based drug design, utilizing three‐dimensional information of receptor structure and ligand‐binding motifs, requires significant amounts of purified protein. To facilitate the purification of CB1, we have expressed the receptor fused to various epitope tags using the baculovirus expression system. In addition, expression levels and ligand‐binding profiles corresponding to the expressed fusion proteins have been compared. C‐terminal histidine (His)‐tagged CB1 gave aBmaxhigher than most other systems previously reported in the literature, and was selected for subsequent metal affinity chromatography purification and mass spectroscopic (MS) analysis. Moreover, cells expressing C‐terminal His‐tagged CB1 were shown to inhibit forskolin‐stimulated cyclic adenosine 3′,5′‐monophosphate (cAMP) production in a concentration‐dependent manner in the presence of CP‐55,940, confirming the expressed receptor's functional characteristics. A Western blot analysis of the purified receptor showed several forms of CB1, the most abundant being a 57 kDa monomeric protein. The purified CB1 preparations were subjected to protein digestion followed by MS. Fragments corresponding to >70% of the receptor were identified by this method, confirming the identity and purity of the expressed protein. The work presented here demonstrates that epitope‐tagged CB1 can be expressed in sufficient amounts and purified to homogeneity for MS analysis. Moreover, these results will serve as a basis for future experiments aimed at characterizing the ligand‐binding domains using covalently reacting receptor probes.
Meiotic chromosome behaviour in an allotriploid derived from an amphidiploid × diploid mating in Cucumis
Authors: Qian C.‐T. , Jahn M. M. , Staub J. E. , Luo X. D. , Chen J. F.
Published: July 2005

A synthetic amphidiploid species Cucumis hyriviis Chen & Kirkbride (2n = 4x = 38; genome designated as HHCC) has recently been created from an interspecific mating between C. sativus L. (2n = 2x = 14: genome designated as CC) and C. hystrx. Chakr. (2n = 2x = 24; genome designated as HH). This amphidiploid is resistant to root knot nematode, tolerant to low irradiance, and has higher nutritional value than standard processing cucumber cultivars. An allotriploid (2n = 3x = 26; HCC) was derived from a cross between C. sativus L. and C. hytivus Chen & Kirkbride. Diploid meiotic behaviour in C. sativus and C. hystrix involves the development of seven and 12 metaphase bivalents respectively. In the derived allotriploid. univalents. bivalents, and trivalents (at relatively low frequency) were observed at metaphase I indicating that some homeologues from the C and H genomes can synapse. Based on a comparative karyotype analysis of cucumber (i.e. chromosome size and pairing behaviour) and aliotriploid plants, the seven bivalents observed at metaphase I were ascertained to be cucumber homologues, while the 12 univalents were of C. hystrix origin thus confirming the allotriploid karyotypic constitution to be HCC. On average, the frequency of trivalents was 0.24 at diakinesis and 0.22 in 100 meiocytes at metaphase I. indicating the possibility of genetic exchange due to the homoeology between the C and H genomes. After simultaneous cytokinesis, only polyads were observed in pollen mother cells (PMCs) at telophase II, which led to the production of sterile pollen grains. Multi‐polarization of chromosomes was dominant at anaphase II. However. in about 20% PMCs. chromosomes separated to form a 7C + 1‐2H complement, suggesting a possible method for the production of alien addition cucumber‐C hystrix lines through further backcrossing of the allotriploid to diploid cucumber.
Inheritance of sesquiterpene carboxylic acid synthesis in crosses of Lycopersicon hirsutum with insect‐susceptible tomatoes
Authors: Frelichowski J. E. , Juvik J. A.
Published: July 2005

Sesquiterpene carboxylic acids (SCA) from Lycopersicon hirsutum f. typicum Humb, & Bonpl, accession LA 1777 confer host‐plant resistance to Helicoverpa zea (Boddie) and Spodoptera exigua (Hübner), two Lepidopteran (Noctuidae) insect pests of the cultivated tomato. L. esculcntum Mill. Hybrids were made between LA 1777 and two accessions of insect‐susceptible tomatoes that do not produce SCA, L. hirsutum LA 1033 and the L. L. esculentum cultivar Chico III, F2 and backcross progeny of both crosses were evaluated for variation in amounts of total SCA in replicated field and greenhouse plantings. Analysis of variance of total SCA in a set of F2 plants of the cross LA 1033 × LA 1777 demonstrated significant variation in SCA attributable to genotype, environment and their interaction. Segregation for high levels of SCA in the populations suggests polygenic inheritance. Broad sense heritability was 0.84, which indicates that phenotypic selection for plants with high SCA is feasible in a backcross‐breeding programme for the introgression of SCA‐mediated insect resistance into tomato cultivars.
Breeding for the improvement of the ideal plant type of sesame
Authors: Baydar H.
Published: July 2005

The objective of this study was to improve the ideal type of sesame, by performing pedigree selection applied to segregating generations of crosses of genotypes with contrasting characters. In the segregating F2 population, the progeny were classified into eight types according to the combinations of carpel number per capsule, capsule number per axil and branching habit. Progeny of individual F2 plants including a particular type were advanced to the F6 generation. It was possible to improve the lines with eight types at the end of the selection process. Some types, especially bicarpels, monocapsule, branch (BMB) and bicarpels, tricapsules, branch (BTB), were considered as ideal plant types in breeding for high‐yielding varieties. While the low‐yielding type quadricarpels, tricapsules, non‐branch (QTN) was the highest in oil content (49.3%), the high‐yielding type BMB was the lowest (43.2%), Although the QTN‐type had the lowest content of oleic acid (41.3%) and the highest content of linoleic acid (43.1 %), the bicarpels, monocapsule, non‐branch (BMN) type had the highest content of oleic acid (48.4%) and the lowest content of linoleic acid (36.6%), Total tocopherol varied between 175.6 and 368.9 mg/kg in the seed oil of the sesame types. The best high‐yielding type BMB was one of the types containing less tocopherol.
Relationships between hard‐seededness and seed weight in mungbean (Vigna radiata) assessed by QTL analysis
Authors: Humphry M. E. , Lambrides C. J. , Chapman S. C. , Aitken E. A. B. , Imrie B. C. , Lawn R. J. , Mcintyre C. L. , Liu C. J.
Published: July 2005

Weather damage reduces the value of commercial mungbean. but hard‐seededness can reduce the level of damage. However. attempts lo breed large‐ and hard‐seeded mungbean varieties have been unsuccessful. To understand the relationship between seed weight and hard‐seededness. these trails were investigated using a quantitative trail loci (QTL) mapping approach with a recombinant inbred population derived from a cross between a completely soft‐seeded variety and a completely hard‐seeded genotype. The two parental genotypes also had a sixfold difference in seed weight. QTL analyses revealed four loci for hard‐seededness and 11 loci for seed weight. Two of the hard‐seeded ness loci co‐localized with seed weight QTL. When seed weight was used as a covariate in the analysis of hard‐seededness from the field data, two of the four hard‐seeded QTL remained significant with the effect al one of these remaining unchanged. These results explain why retaining hard‐seededness in large seeded mungbean lines has been unsuccessful. The existence of a persistent locus, however. indicated that breeding large and persistently hard‐seeded varieties of mungbean may be possible.
Development of cytoplasmic‐genic male sterility in safflower
Authors: Anhani K.
Published: July 2005

An interspecific cross was made between Carthamaus oxyacantha and the cultivated species C. tinctorius to develop a cytoplasmic‐genic male sterility (CMS) system in safflower. C. oxyacantha was the donor of sterile cytoplasm. The 3: 1 segregation pattern observed in BC1F2 suggested single gene control with dominance of male‐fertility over male‐sterility. The information obtained from crossing male sterile X male fertile plants in BC1F3 and BC1F4 generations showed statistically significant single gene (1: 1) segregation for male sterility vs. male fertility. The results demonstrated that C. tinctorius possesses a nuclear fertility restorer gene and that a single dominant allele restored fertility (Rf) in progeny carrying CMS cytoplasm of C. oxyacantha. Male sterility occurred with the homozygous recessive condition (rfrf) in a sterile C. oxyacantha cytoplasm background and not in the normal cytoplasm of C. tinctorius. The genetic background of different restorer lines of C. tinctorius having normal cytoplasm did not effect fertility restoration. The absence of male sterile plants in C. tinctorius populations ruled out the possibility of genetic male sterility. Normal meiosis in F1 and BC1F2 ruled out a cytogenetic basis for the occurrence of male sterility.
Genetic variation between Spanish and American versions of sweet corn inbred lines
Authors: Revilla P. , Abuin M. C. , Malvar R. A. , Soengas P. , Ordás B. , Ordás A.
Published: July 2005

Conservation of maize inbred lines in different stations causes variability among strains. The objective of this research was to determine agronomic and molecular differences in American sweet corn inbreds maintained in Spain. American and Spanish strains of five sweet corn inbred lines were characterized by using 34 RAPD primers that produced 168 consistent bands. Strains of four of these inbreds were crossed in a diallel design, and hybrids were evaluated in four environments in northwestern Spain. The RAPD characterization showed no differences between strains for two inbreds, while divergence between strains was largest for the inbred 15125. Most primers did not reveal any variability between pairs of strains, while some primers produced variations at high rates. Differences in agronomic performance among Spanish and American strains were most important for P51, followed by P39, while strains 1453 or 15125 were not significantly different. Molecular differences between strains were not related to agronomic performance. Residual heterozygosity or outcrossing cannot explain these results. Lack of adaptation could have caused stress‐induced mutagenesis. Natural selection could have eliminated unfavourable selective mutations, but neutral mutations can be found at the molecular level and favourable mutations could have been selected at the agronomic level.
Chromosomal location of powdery mildew resistance gene Pm16 in wheat using SSR marker analysis
Authors: Chen X. M. , Luo Y. H. , Xia X. C. , Xia L. Q. , Chen X. , Ren Z. L. , He Z. H. , Jia J. Z.
Published: July 2005

The use of resistant cultivars is a most economical way to control powdery mildew (Blumeria graminis f.sp. tritici) in wheat (Triticum aestivum L.). Identification of molecular markers closely linked to resistance genes can greatly increase the efficiency of pyramiding resistance genes in wheat cultivars. The objective of this study was to identify molecular markers closely linked lo the powdery mildew resistance gene Pm16. An F2 population with 156 progeny was produced from the cross‘Chancellor’(susceptible) ×‘70281’ (resistant), A total of 45 SSR markers on chromosomes 4A and 5B of wheat and 15 SSRs on chromosome 3 of rice was used lo lest the parents, as well as the resistant and susceptible bulks: the resulting polymorphic markers were used to genotype the F2 progeny. Results indicated that the SSR marker Xgwm159, located on the short arm of chromosome 5B, is closely linked to Pm16 (genetic distance: 5.3 CM). The cytogenetical data presented in an original report, in combination with this molecular analysis, suggests that Pm16 may he located on a translocated 4A.5BS chromosome.
Response to Mycosphaerella pinodes in a germplasm collection of Pisum spp
Authors: Fondevilla S. , ÅVila C. M. , Cubero J. I. , Rubiales D.
Published: July 2005

Little resistance against Mycosphaerella pinodes is available in pea. In this work 78 accessions of Pisum were screened for resistance to M. pinodes. Fourteen accessions showed a good level of resistance in seedlings under controlled conditions, and in mature plants in the field. The highest levels of resistance were found in P. fuivum, followed by P. sativum ssp. eiatius and P. sativum ssp. syriacum. Resistance of five selected accessions was effective against different isolates of M. pinodes originating from different countries. Resistant accessions reported in this paper have been successfully hybridized with field pea cultivars.
Usefulness of the addition of metformin to insulin in pediatric patients with type 1 diabetes mellitus
Authors: Urakami Tatsuhiko , Morimoto Shigeo , Owada Misao , Harada Kensuke
Published: July 2005

Background
: The aim of this study was to evaluate the effect of metformin in addition to insulin therapy in adolescents and young adults with type 1 diabetes mellitus.

Methods
: Nine patients, two males and seven females, aged 18.1 ± 3.0 years, with type 1 diabetes mellitus were studied. They were relatively overweight with a body mass index (BMI) of 24.2 ± 1.8 and had high levels of HbA1c at 9.5 ± 1.2% despite high doses of insulin of 74.0 ± 31.2 U/day. Metformin at the dose of 500–750 mg daily was administered to the patients in addition to insulin therapy for 1 year.

Results
: HbA1c, BMI and insulin dose were compared before 1 year without metformin therapy, at baseline, and at 3, 6 and 12 months during the use of metformin in addition to insulin therapy. HbA1c lowered (8.6 ± 1.4**, 8.4 ± 1.3**, 8.4 ± 1.2*%), BMI was reduced (23.9 ± 1.7*, 23.8 ± 1.8, 23.5 ± 1.8*), and insulin requirement decreased (69.8 ± 29.7*, 68.7 ± 29.8**, 67.3 ± 29.1**U/d) significantly after the start of metformin therapy (*P < 0.05, **P < 0.01 vs at baseline). There were no adverse events, not even lactic acidosis, during the study period.

Conclusion
: Metformin is safe and may represent a useful adjunct to the management of type 1 diabetes mellitus in adolescents and young adults who have poor glycemic control despite a large amount of insulin.
Is leptin a predictive factor in the end of therapy response in chronic hepatitis B?
Authors: Selimo}lu Mukadder Ayle , Ertekin Vildan
Published: July 2005

Background
: The purpose of this study was to determine the leptin levels in children with chronic hepatitis B virus (HBV) infection, and to evaluate the effect of serum leptin levels on the end of therapy response (ETR). It is known that leptin stimulates T‐cell immunity and so T‐cell mediated immune response is critical in the outcome of chronic HBV infection.

Methods
: Leptin levels in children with chronic HBV infection were investigated and its effects on the ETR in 24 children who were treated with interferon‐α and lamivudine combination therapy were evaluated.

Results
: The mean leptin level of the patients was higher than that of healthy children (P = 0.034). Of the patients, seven (29.2%) had ETR. The mean hepatic activity index and portal inflammation score were higher, the HBV DNA was lower, and the leptin level was similar in children with ETR when compared to others (P = 0.017, P = 0.04, P = 0.007, P = 0.34, respectively). HBV DNA and the fibrosis score were positively correlated (P = 0.016).

Conclusion
: Although the higher leptin value observed in children with ETR was not statistically significant, because of close interactions between leptin, cytokines and lymphocytes, it is thought that leptin should be investigated as a predictive factor of ETR in further studies.
Cross‐cultural comparison of the neurobehavioral characteristics of Chinese and Japanese neonates
Authors: Loo Kek Khee , Ohgi Shohei , Zhu Honglin , Howard Judy , Chen Lian
Published: July 2005

Background
: Similarities and differences in the neurobehavioral repertoire of neonates from different cultures have been noted using the Brazelton Neonatal Behavioral Assessment Scale (NBAS). By identifying the behavioral attributes of newborns that are seen more universally versus those that are culture‐specific, comparative studies increase understanding of the roles of cultural factors in shaping the behavioral trajectory of infants. This study compared the neurobehavioral characteristics of neonates in Chengdu, China and Nagasaki, Japan.

Methods
: The Brazelton NBAS was administered within the first week of life to 40 Chinese and 62 Japanese healthy, full‐term neonates in Chengdu and Nagasaki.

Results
: The two groups differed significantly in birthweight, gender, age at time of exam, mode of delivery, and gestational age. When these variables, in addition to group status (Chinese or Japanese), were entered into multiple linear analyses with NBAS cluster scores as dependent variables, group status independently predicted Range of State and Habituation scores. The group status did not predict Orientation, Motor, Regulation of States, Autonomic Stability, and Abnormal Reflexes cluster scores.

Conclusion
: These results suggest that Japanese newborns habituated more readily to stimuli, and were less irritable than newborns in the Chinese sample. The infants in the two groups shared many similar neurobehavioral characteristics as well. The implications of these similarities and differences were discussed.
Individualisation, Choice and Structure: A Discussion of Current Trends in Sociological Analysis
Authors: Brannen Julia , Nilsen Ann
Published: July 2005

In this paper we seek to explore a tendency in current sociological thought to highlight notions of choice and autonomy in writings about contemporary Western societies. We wish to draw attention to some of the consequences of leaving out discussions of the structural aspects of societies and people's lives, for individuals as well as for the development and application of sociological theory and its ability to understand the connection between history and individual biography. Our discussion is based on qualitative research that we have conducted in recent years, and draws on focus groups with young people in Norway and Britain. From this critique we seek to demonstrate how concepts that take account of context and structure as well individual subjectivities can create a better ‘fit’ with complex and diverse realties.
South Asian Disabled Women: Negotiating Identities
Authors: Hussain Yasmin
Published: July 2005

This paper is concerned with the identities of disabled South Asian women within Britain. It presents empirical evidence concerning how disability, gender and ethnicity are negotiated simultaneously for young disabled Muslim and Sikh women. How these identities are negotiated is analysed in the realms of family, religion and marriage drawing on qualitative interviews with the young women, their parents and siblings. The paper argues against ideas of singular identity or the hierarchisation of identities or oppressions. The paper contributes to contemporary debates about how young South Asian women are constructing new forms of identity in Britain.
Proteasome inhibition by lactacystin in primary neuronal cells induces both potentially neuroprotective and pro‐apoptotic transcriptional responses: a microarray analysis
Authors: Yew Elaine Hau Jin , Cheung Nam Sang , Choy Meng Shyan , Qi Robert Z. , Lee Alan Yiu‐Wah , Peng Zhao Feng , Melendez Alirio J. , Manikandan Jayapal , Koay Evelyn Siew‐Chuan , Chiu Lily‐Lily , Ng Wooi Loon , Whiteman Matthew , Kandiah Jeyaseelan , Halliwell Barry
Published: July 2005

Although inhibition of the ubiquitin proteasome system has been postulated to play a key role in the pathogenesis of neurodegenerative diseases, studies have also shown that proteasome inhibition can induce increased expression of neuroprotective heat‐shock proteins (HSPs). The global gene expression of primary neurons in response to treatment with the proteasome inhibitor lactacystin was studied to identify the widest range of possible pathways affected. Our results showed changes in mRNA abundance, both at different time points after lactacystin treatment and at different lactacystin concentrations. Genes that were differentially up‐regulated at the early time point but not when most cells were undergoing apoptosis might be involved in an attempt to reverse proteasome inhibitor‐mediated apoptosis and include HSP70, HSP22 and cell cycle inhibitors. The up‐regulation of HSP70 and HSP22 appeared specific towards proteasome inhibitor‐mediated cell death. Overexpression of HSP22 was found to protect against proteasome inhibitor‐mediated loss of viability by up to 25%. Genes involved in oxidative stress and the inflammatory response were also up‐regulated. These data suggest an initial neuroprotective pathway involving HSPs, antioxidants and cell cycle inhibitors, followed by a pro‐apoptotic response possibly mediated by inflammation, oxidative stress and aberrant activation of cell cycle proteins.
Differential effects ofl‐DOPA on monoamine metabolism, cell survival and glutathione production in midbrain neuronal‐enriched cultures from parkin knockout and wild‐type mice
Authors: Casarejos M. J. , Solano R. M. , Menéndez J. , Rodríguez‐Navarro J. A. , Correa C. , García de Yébenes J. , Mena M. A.
Published: July 2005

l‐DOPA is the most effective treatment for Parkinson's disease but in isolated neuronal cultures it is neurotoxic for dopamine (DA) neurones. Experimentsin vivoand clinical studies have failed to show toxicity ofl‐DOPA in animals or patients but that does not exclude the possibility of a toxic effect ofl‐DOPA on patients with certain genetic risk factors. Mutations of the parkin gene are the most frequent cause of hereditary parkinsonism. Parkin null mice have a mild phenotype that could be modified by different neurotoxins. The aim of this study was to investigate whether the toxic effects ofl‐DOPA on DA neurones are amplified in parkin null mice. We have measured the effects ofl‐DOPA on cell viability, tyrosine hydroxylase (TH) expression, DA metabolism and glutathione levels of parkin knockout (PK‐KO) midbrain cultures. Neuronal‐enriched cultures from PK‐KO mice have similar proportions of the different cell types with the exception of a significant increment of microglial cells.l‐DOPA (400 µmfor 24 h) reduced the number of TH‐immunoreactive cells to 50% of baseline and increased twofold the percentage of apoptotic cells in cultures of wild‐type (WT) animals. The PK‐KO mice, however, are not only resistant to thel‐DOPA‐induced pro‐apoptotic effects but they have an increased number of TH‐immunoreactive neurones after treatment withl‐DOPA, suggesting thatl‐DOPA is toxic for neurones of WT mice but not those of parkin null mice. MAPK and phosphatidylinositol‐3 kinase signalling pathways are not involved in the differentiall‐DOPA effects in WT and PK‐KO cultures. Intracellular levels ofl‐DOPA were not different in WT and parkin null mice but the intracellular and extracellular levels of DA and 3‐4‐dihydroxyphenylacetic acid, however, were significantly increased in parkin null animals. Furthermore, monoamine oxidase activity was significantly increased in parkin null mice, suggesting that these animals have an increased metabolism of DA. The levels of glutathione were further increased in parkin null mice than in controls both with and without treatment withl‐DOPA, suggesting that a compensatory mechanism may protect DA neurones from neuronal death. This study opens new avenues for understanding the mechanisms of action ofl‐DOPA on DA neurones in patients with Park‐2 mutations.
Inhibition of Rac GTPase triggers a c‐Jun‐ and Bim‐dependent mitochondrial apoptotic cascade in cerebellar granule neurons
Authors: Le Shoshona S. , Loucks F. Alexandra , Udo Hiroshi , Richardson‐Burns Sarah , Phelps Reid A. , Bouchard Ron J. , Barth Holger , Aktories Klaus , Tyler Kenneth L. , Kandel Eric R. , Heidenreich Kim A. , Linseman Daniel A.
Published: July 2005

Rho GTPases are key transducers of integrin/extracellular matrix and growth factor signaling. Although integrin‐mediated adhesion and trophic support suppress neuronal apoptosis, the role of Rho GTPases in neuronal survival is unclear. Here, we have identified Rac as a critical pro‐survival GTPase in cerebellar granule neurons (CGNs) and elucidated a death pathway triggered by its inactivation. GTP‐loading of Rac1 was maintained in CGNs by integrin‐mediated (RGD‐dependent) cell attachment and trophic support. Clostridium difficile toxin B (ToxB), a specific Rho family inhibitor, induced a selective caspase‐mediated degradation of Rac1 without affecting RhoA or Cdc42 protein levels. Both ToxB and dominant–negative N17Rac1 elicited CGN apoptosis, characterized by cytochrome c release and activation of caspase‐9 and ‐3, whereas dominant–negative N19RhoA or N17Cdc42 did not cause significant cell death. ToxB stimulated mitochondrial translocation and conformational activation of Bax, c‐Jun activation, and induction of the BH3‐only protein Bim. Similarly, c‐Jun activation and Bim induction were observed with N17Rac1. A c‐jun N‐terminal protein kinase (JNK)/p38 inhibitor, SB203580, and a JNK‐specific inhibitor, SP600125, significantly decreased ToxB‐induced Bim expression and blunted each subsequent step of the apoptotic cascade. These results indicate that Rac acts downstream of integrins and growth factors to promote neuronal survival by repressing c‐Jun/Bim‐mediated mitochondrial apoptosis.
Celastrol protects against MPTP‐ and 3‐nitropropionic acid‐induced neurotoxicity
Authors: Cleren Carine , Calingasan Noel Y. , Chen Junya , Beal M. Flint
Published: July 2005

Oxidative stress and inflammation are implicated in neurodegenerative diseases including Parkinson's disease (PD) and Huntington's disease (HD). Celastrol is a potent anti‐inflammatory and antioxidant compound extracted from a perennial creeping plant belonging to theCelastraceaefamily.Celastrol is known to prevent the production of proinflammatory cytokines, inducible nitric oxide synthase and lipid peroxidation. Mice were treated with celastrol before and after injections of MPTP, a dopaminergic neurotoxin, which produces a model of PD. A 48% loss of dopaminergic neurons induced by MPTP in the substantia nigra pars compacta was significantly attenuated by celastrol treatment. Moreover, celastrol treatment significantly reduced the depletion in dopamine concentration induced by MPTP. Similarly, celastrol significantly decreased the striatal lesion volume induced by 3‐nitropropionic acid, a neurotoxin used to model HD in rats. Celastrol induced heat shock protein 70 within dopaminergic neurons and decreased tumor necrosis factor‐α and nuclear factor κ B immunostainings as well as astrogliosis. Celastrol is therefore a promising neuroprotective agent for the treatment of PD and HD.