bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel mutations associated with metachromatic leukodystrophy: Phenotype and expression studies in nine Czech and Slovak patients
Authors: Berná Linda , Gieselmann Volkmar , Poupětová Helena , Hřebíček Martin , Elleder Milan , Ledvinová Jana
Published: May 2004

Metachromatic leukodystrophy (MLD) is an inherited demyelinating disorder caused by the deficiency of arylsulphatase A (ASA). This defect leads to an accumulation of galactosylceramide I3‐sulphates (sulphatides) in lysosomes of different tissues. We report on mutations found in a group of nine patients from the Czech and Slovak Republics (former Czechoslovakia). Their diagnosis was confirmed by determination of the activity of arylsulphatase A in leukocytes and by abnormal urinary excretion of sulphatides. All alleles of the patients were identified and eight different mutations were found. They include four novel missense mutations in one infantile (D29N), one juvenile (C294Y), and three adult (C156R, G293S) patients. Four mutations were previously described sequence alterations (459 + 1G > A, G309S, I179S, and P426L). Polymorphisms characteristic for the ASA pseudodeficiency allele were not found in the patients. Substitutions of D29N, C294Y, and G293S in arylsulphatase A caused a severe reduction of enzyme activity in transient expression studies. In contrast, the C156R substitution reduces arylsulphatase A only to 50% of wild type ASA activity. Since no other mutations were found in this patient, the contribution of this mutation to the development of disease remains unclear. © 2004 Wiley‐Liss, Inc.
Mitochondrial DNA polymorphisms and extraversion
Authors: Kato Chieko , Umekage Tadashi , Tochigi Mamoru , Otowa Takeshi , Hibino Hiroyuki , Ohtani Toshiyuki , Kohda Kazuhisa , Kato Nobumasa , Sasaki Tsukasa
Published: May 2004

Mitochondria is the major site of energy production in cells, therefore, mitochondrial abnormality may affect functions of organs including the brain, which constantly requires high levels of energy consumption. Previous studies have suggested a role of mitochondria and their DNA polymorphisms in neuro‐psychiatric disorders, including Alzheimer's disease, Parkinson's disease, schizophrenia and bipolar mood disorder. Thus, we hypothesized that mitochondrial DNA polymorphisms might be related with the development of personality. The present study investigated a role of two mitochondrial DNA polymorphisms, the C5178A and A10398G, in personality traits evaluated using the NEO PI‐R scores in 238 healthy Japanese volunteers. Subjects with the 5178A genotype showed significantly higher extraversion score than those with the 5178C genotype (P = 0.027), while no significant association was observed between the C5178A polymorphism and other scores. No significant association was found between the A10398G polymorphism and any scores. Regarding the 5178–10398 haplotype, the score of extraversion, not other scores, was significantly associated with the A–G haplotype (P = 0.042). Although further studies are recommended for the confirmation, the result may suggest a role of the mitochondrial DNA polymorphism in the personality trait. © 2004 Wiley‐Liss, Inc.
MECP2 structural and 3′‐UTR variants in schizophrenia, autism and other psychiatric diseases: A possible association with autism
Authors: Shibayama Akane , Cook Edwin H. , Feng Jinong , Glanzmann Cecile , Yan Jin , Craddock Nick , Jones Ian R. , Goldman David , Heston Leonard L. , Sommer Steve S.
Published: May 2004

Mutations in the gene coding for methyl‐CpG‐binding protein 2 (MECP2) cause Rett syndrome (RTT) and have also been reported in a number of X‐linked mental retardation syndromes. Furthermore, putative mutations recently have been described in a few autistic patients and a boy with language disorder and schizophrenia. In this study, DNA samples from individuals with schizophrenia and other psychiatric diseases were scanned in order to explore whether the phenotypic spectrum of mutations in the MECP2 gene can extend beyond the traditional diagnoses of RTT in females and severe neonatal encephalopathy in males. The coding regions, adjacent splicing junctions, and highly conserved segments of the 3′‐untranslated region (3′‐UTR) were examined in 214 patients, including 106 with schizophrenia, 24 with autism, and 84 patients with other psychiatric diseases by detection of virtually all mutations‐single strand conformation polymorphism (SSCP) (DOVAM‐S). To our knowledge, this is the first analysis of variants in conserved regions of the 3′‐UTR of this gene. A total of 5.2 kb per haploid gene was analyzed (1.5 Mb for 214 patients). A higher frequency of missense and 3′‐UTR variants was found in autism. One missense and two 3′‐UTR variants were found in 24 patients with autism versus one patient with a missense change in 144 ethnically similar individuals without autism (P = 0.009). These mutations suggest that a possible association between MECP2 mutations and autism may warrant further study. © 2004 Wiley‐Liss, Inc.
Association analysis of the DRD4 and COMT genes in methamphetamine abuse
Authors: Li Tao , Chen Chi‐ken , Hu Xun , Ball David , Lin Shih‐Ku , Chen Wai , Sham Pak C. , Loh El‐Wui , Murray Robin M. , Collier David A.
Published: May 2004

We analyzed two polymorphisms in genes encoding proteins of the dopamine system, the Val158Met polymorphism in the catechol‐O‐methyltransferase gene and the 120‐bp VNTR polymorphism in the promoter of the dopamine D4 receptor gene for association with methamphetamine abuse. We used a case/control design with 416 methamphetamine abusing subjects and 435 normal controls. All subjects were Han Chinese from Taiwan. We found an excess of the high activity Val158 allele in the methamphetamine abuser group, consistent with several previous reports of association of this allele with drug abuse. The 120‐bp VNTR polymorphism in the promoter of the dopamine D4 receptor gene itself did not show significant association with methamphetamine abuse. However, analysis of the 120‐bp VNTR polymorphism and the exon 3 VNTR in the dopamine D4 receptor as a haplotype showed significant association with methamphetamine abuse, which gave an empirical P value 0.0034 for a heterogeneity model. Moreover, there were significant interactive effects between polymorphisms in the catechol‐O‐methyltransferase and dopamine D4 genes. The evidence of interaction between COMT 158 Val/Met and DRD4 48‐bp VNTR polymorphisms (P = 0.0003, OR = 1.45, 95% CI: 1.148–1.77), and between COMT 158 Val/Met and DRD4 120 bp promoter polymorphisms (P = 0.01, OR = 1.10, 95% CI: 1.10–1.18) were significant but the latter was weak. We conclude that genetic variation in the dopamine system may encode an additive effect on risk of becoming a methamphetamine abuser. © 2004 Wiley‐Liss, Inc.
Genetic disorders affecting white matter in the pediatric age
Authors: Di Rocco Maja , Biancheri Roberta , Rossi Andrea , Filocamo Mirella , Tortori‐Donati Paolo
Published: May 2004

Pediatric white matter disorders can be distinguished into well‐defined leukoencephalopathies, and undefined leukoencephalopathies. The first category may be subdivided into: (a) hypomyelinating disorders; (b) dysmyelinating disorders; (c) leukodystrophies; (d) disorders related to cystic degeneration of myelin; and (e) disorders secondary to axonal damage. The second category, representing up to 50% of leukoencephalopathies in childhood, requires a multidisciplinar approach in order to define novel homogeneous subgroups of patients, possibly representing “new genetic disorders” (such as megalencephalic leukoencepahlopathy with subcortical cysts and vanishing white matter disease that have recently been identified). In the majority of cases, pediatric white matter disorders are inherited diseases. An integrated description of the clinical, neuroimaging and pathophysiological features is crucial for categorizing myelin disorders and better understanding their genetic basis. A review of the genetic disorders affecting white matter in the pediatric age, including some novel entities, is provided. © 2004 Wiley‐Liss, Inc.
Linkage of eye movement dysfunction to chromosome 6p in schizophrenia: Additional evidence
Authors: Matthysse Steven , Holzman Philip S. , Gusella James F. , Levy Deborah L. , Harte Christopher B. , Jørgensen Åge , Møller Lise , Parnas Josef
Published: May 2004

Establishing the genetics of physiological traits associated with schizophrenia may be an important first step in building a neurobiological bridge between the disease phenotype and its genetic underpinnings. One of the best known of the traits associated with schizophrenia is a disorder of smooth pursuit eye tracking (ETD), which is present in 50–80% of schizophrenia patients. ETD is more than three times more prevalent in the families of a schizophrenia patient than is schizophrenia itself. Arolt et al. [1999] estimated LOD scores for ETD of 2.85 for D6S282 and 3.70 for D6S271, two markers on 6p21.1, as well as obtaining an indication of possible linkage for schizophrenia. Our sample comprised two large families in Denmark. Markers in the region that was implicated by the study of Arolt et al. [1996, 1999] were analyzed as part of a genome scan using the “latent trait (L.T.) model” for the co‐transmission of schizophrenia and ETD that we had previously fitted to segregation analysis data from Norway. We obtained a LOD score of 2.05 for D6S1017, a marker within 3 cM of the positive markers obtained by Arolt et al. [1996, 1999]. We regard our results as independent evidence supporting the findings of Arolt et al. [1996, 1999] and also as support for the L.T. model as a way of combining the traits ETD and schizophrenia. © 2004 Wiley‐Liss, Inc.
Association analysis of two candidate phospholipase genes that map to the chromosome 15q15.1‐15.3 region associated with reading disability
Authors: Morris D.W. , Ivanov D. , Robinson L. , Williams N. , Stevenson J. , Owen M.J. , Williams J. , O'Donovan M.C.
Published: May 2004

Molecular genetic studies have suggested a reading disability (RD, dyslexia) susceptibility locus on chromosome 15q. We have previously mapped this locus by association to the region surrounding D15S994. Very little is known about the neurobiological processes involved in RD, and therefore selecting positional candidate genes for analysis based upon function is difficult. Nevertheless we were able to identify two functional candidates based upon existing hypotheses. Both were phospholipase genes, phospholipase C β 2 (PLCB2) and phospholipase A2, group IVB (cytosolic; PLA2G4B). D15S944 is located within PLCB2 and is 1.6 Mb from PLA2G4B. We examined each gene for association using a mixed direct and indirect association approach, a case (n = 164)/control (n = 174) sample, and a partially overlapping sample of 178 RD parent‐proband trios from South Wales and England. Mutation analysis revealed 14 sequence variants in PLCB2 and 33 variants in PLA2G4B. All non‐synonymous SNPs were genotyped as were SNPs across each gene with maximum distance between SNPs of 6 kb. Case‐control analyses revealed modest evidence (0.01 < P < 0.05) for association between a single variant in PLCB2 and two variants in PLA2G4B. However, association was not confirmed in the family based sample. As the latter sample has previously generated replicated significant evidence for association between RD and markers/haplotypes surrounding D15S944, it should have sufficient power to detect association to variants in susceptibility gene itself. We conclude that neither gene accounts for the association signal we previously observed. As these are the only clear cut functional candidate genes in the region, identification of the putative susceptibility locus for RD on 15q will require more methodical non‐hypothesis driven positional cloning approaches. © 2004 Wiley‐Liss, Inc.
Dermatoglyphic profile in 22q deletion syndrome
Authors: Martín B. , Fañanás L. , Gutiérrez B. , Chow E.W.C. , Bassett A.S.
Published: May 2004

A genetic subtype of schizophrenia has been described in 22q11 Deletion syndrome. Previous studies have described an excess of dermatoglyphic alterations in schizophrenia, such as low a–b ridge counts (ABRCs), a high frequency of ridge dissociations, and increased dermatoglyphic fluctuating asymmetry. Little is known however, about the dermatoglyphic profile of 22qDS subjects showing psychotic symptoms and its similarity to the previously reported anomalies in schizophrenia. We studied the palmar dermatoglyphics of 22 subjects with 22qDS of predominantly Caucasian origin, 15 of whom had psychotic illness, and in 84 healthy controls of similar ethnicity. We observed higher values for total ATD angle in cases than in controls (P = 0.04). In addition, there was an excess of radial figures in the hypothenar area in cases, especially in the left hand. Interestingly, greater fluctuating asymmetry, determined by the absolute difference between right and left ABRC, was observed in 22qDS subjects compared to controls (P = 0.05). However, no differences were found for ABRCs and frequency of dissociations. Despite the small sample size, the palmprints analyzed suggest the existence of an altered dermatoglyphic profile in 22qDS, involving: (i) ATD angle amplitude, (ii) presence of radial loops in the hypothenar area, and (iii) an increment of fluctuating asymmetry. The first two features are similar to those found in other genetic syndromes associated with low IQ, while high levels of fluctuating asymmetry have often been reported in schizophrenia. © 2004 Wiley‐Liss, Inc.
Are phonological processing deficits part of the broad autism phenotype?
Authors: Bishop Dorothy V.M. , Maybery Murray , Wong Dana , Maley Alana , Hill Wayne , Hallmayer Joachim
Published: May 2004

Two tests of phonological processing, nonword repetition, and nonsense passage reading, were administered to 80 probands with autistic disorder or PDDNOS (index cases) and 59 typically developing controls, together with their parents and siblings. In addition, parents completed a questionnaire about history of language and literacy problems, and all participants were given tests of verbal (VIQ) and performance IQ (PIQ). Parents also completed the Autism‐Spectrum Quotient, which was used to index the broad autism phenotype. Index probands scored well below control probands on the two phonological tests. However, on neither phonological measure did index relatives differ from control relatives. Within the index group, there was no relationship between the proband's level of VIQ, or age at achieving phrase speech, and phonological score of relatives. VIQ was the only measure to show any familiality within the index group. Reported history of language and literacy problems did not differentiate index parents from control parents overall, but those who were categorized as cases of the broad phenotype reported more history of language and literacy problems than did other index parents. However, they did not have poorer scores on the phonological measures. It is concluded that phonological processing deficits are not part of the broad autism phenotype. © 2004 Wiley‐Liss, Inc.
Conformational basis for the biological activity of TOAC‐labeled angiotensin II and bradykinin: Electron paramagnetic resonance, circular dichroism, and fluorescence studies
Authors: Schreier Shirley , Barbosa Simone R. , Casallanovo Fábio , Vieira Renata de F. F. , Cilli Eduardo M. , Paiva Antonio C. M. , Nakaie Clóvis R.
Published: May 2004

N‐Terminally and internally labeled analogues of the hormones angiotensin (AII, DRVYIHPF) and bradykinin (BK, RPPGFSPFR) were synthesized containing the paramagnetic amino acid 2,2,6,6‐tetramethylpiperidine‐1‐oxyl‐4‐amino‐4‐carboxylic acid (TOAC). TOAC replaced Asp1 (TOAC1‐AII) and Val3 (TOAC3‐AII) in AII and was inserted prior to Arg1 (TOAC0‐BK) and replacing Pro3 (TOAC3‐BK) in BK. The peptide conformational properties were examined as a function of trifluoroethanol (TFE) content and pH. Electron paramagnetic resonance spectra were sensitive to both variables and showed that internally labeled analogues yielded rotational correlation times (τC) considerably larger than N‐terminally labeled ones, evincing the greater freedom of motion of the N‐terminus. In TFE, τC increased due to viscosity effects. Calculation of τCpeptide/τCTOAC ratios indicated that the peptides acquired more folded conformations. Circular dichroism spectra showed that, except for TOAC1‐AII in TFE, the N‐terminally labeled analogues displayed a conformational behavior similar to that of the parent peptides. In contrast, under all conditions, the TOAC3 derivatives acquired more restricted conformations. Fluorescence spectra of AII and its derivatives were especially sensitive to the ionization of Tyr4. Fluorescence quenching by the nitroxide moiety was much more pronounced for TOAC3‐AII. The conformational behavior of the TOAC derivatives bears excellent correlation with their biological activity, since, while the N‐terminally labeled peptides were partially active, their internally labeled counterparts were inactive [Nakaie, C. R., et al., Peptides 2002, 23, 65–70]. The data demonstrate that insertion of TOAC in the middle of the peptide chain induces conformational restrictions that lead to loss of backbone flexibility, not allowing the peptides to acquire their receptor‐bound conformation. © 2004 Wiley Periodicals, Inc. Biopolymers, 2004
Ribosome motions modulate electrostatic properties
Authors: Trylska Joanna , Konecny Robert , Tama Florence , Brooks Charles L. , McCammon J. Andrew
Published: May 2004

The electrostatic properties of the 70S ribosome of Thermus thermophilus were studied qualitatively by solving the Poisson–Boltzmann (PB) equation in aqueous solution and with physiological ionic strength. The electrostatic potential was calculated for conformations of the ribosome derived by recent normal mode analysis (Tama, F., et al. Proc Natl Acad Sci USA 2003 100, 9319–9323) of the ratchet‐like reorganization that occurs during translocation (Frank, J.; Agrawal, R. K. Nature 2000 406, 318–322). To solve the PB equation, effective parameters (charges and radii), applicable to a highly charged backbone model of the ribosome, were developed. Regions of positive potential were found at the binding site of the elongation factors G and Tu, as well as where the release factors bind. Large positive potential areas are especially pronounced around the L11 and L6 proteins. The region around the L1 protein is also positively charged, supporting the idea that L1 may interact with the E‐site tRNA during its release from the ribosome after translocation. Functional rearrangement of the ribosome leads to electrostatic changes which may help the translocation of the tRNAs during the elongation stage. © 2004 Wiley Periodicals, Inc. Biopolymers, 2004
One‐step analysis of testis steroidogenesis from neonatal exposure to synthetic estrogen by normal‐phase HPLC
Authors: Kuwada Masahiro , Kawashima Rei , Furudate Senich , Sugano Sachiko , Maki Jun
Published: May 2004

Neonatal exposure to synthetic estrogen endocrine disruptors or estrogen‐receptor inhibitors induces developmental abnormalities in the male reproductive system. To investigate whether neonatal exposure affects spermatogenesis in juvenile and pubertal testis, Sprague–Dawley rat pups were given synthetic estrogen endocrine disruptors or estrogen‐receptor inhibitors by a single injection on the day of birth at concentrations ranging between 2 to 40 mm, and sacriﬁced on day 21 (juvenile), 35 (prepuberty) or 50 (puberty). The testes were weighed and examined histologically at each stage. Further, the metabolites of steroidogenesis were analyzed using normal‐phase high performance liquid chromatography. Neonatal exposure signiﬁcantly reduced testis weights and steroidogenesis to one‐ﬁfth to one‐half of that of the juvenile control, and further suppressed irreversible steroidogenesis and spermatogenesis during puberty. Copyright © 2004 John Wiley & Sons, Ltd.
Encapsulation of isohexenylnaphthazarins in cyclodextrins
Authors: Assimopoulou A. N. , Papageorgiou V. P.
Published: May 2004

Naturally occurring isohexenylnaphthazarins (IHN), such as Alkannin, Shikonin (A/S) and their derivatives, are potent pharmaceutical substances with a wide spectrum of biological activity. In the present study, inclusion complexes of alkannin and shikonin commercial samples and IHN derivatives in the form of an oily extract of Alkanna tinctoria roots were formed with β‐cyclodextrin (CD) and β‐HPCD. These complexes were investigated to evaluate the effect of complexation on their aqueous solubility, decoloration, and also the percentage of polymeric A/S and IHN derivatives enclosed in the CDs cavity, since these decrease the active monomeric IHN. Both β‐CD and β‐HPCD increased the aqueous solubility of A/S and IHN derivatives and thus inclusion complexes can be used as drug delivery systems for A/S in both internal (capsules, tablets) and external hydrophilic pharmaceutical and cosmetic preparations (creams, gels, sprays) with enhanced bioavailability. The inclusion complexes formed had a pale purple colour, contributing to the partial decoloration of the A/S and thus of the ﬁnal pharmaceutical preparations. Finally, CDs selectively included more monomeric and less polymeric IHN, compared with the initial each time sample that is encapsulated; thus inclusion complexes may present enhanced biological activity. Copyright © 2004 John Wiley & Sons, Ltd.
Simultaneous determination of chlorpheniramine and pseudoephedrine in human plasma by liquid chromatography–tandem mass spectrometry
Authors: Chen Xiaoyan , Zhang Yong , Zhong Dafang
Published: May 2004

A sensitive and speciﬁc procedure for simultaneous quantitation of chlorpheniramine and pseudoephedrine in human plasma has been developed and validated. Analytes were extracted from plasma samples by liquid–liquid extraction, separated on a Diamonsil C18 column (250 × 4.6 mm i.d.) and detected by tandem mass spectrometry with an atmospheric pressure chemical ionization interface. Diphenhydramine was used as the internal standard. The method has a lower limit of quantitation of 0.2 and 2.0 ng/mL for chlorpheniramine and pseudoephedrine, respectively. The intra‐ and inter‐day relative standard deviation, calculated from quality control (QC) samples were below 4.3% for chlorpheniramine and below 9.5% for pseudoephedrine. The inter‐day relative error as determined from QC samples was within 4.7% for each analyte. The overall extraction recoveries of chlorpheniramine and pseudoephedrine were 77 and 61% on average, respectively. The method was successfully applied to pharmaockinetic study of chlorpheniramine and pseudoephedrine in volunteers receiving formulations containing 4 mg of chlorpheniramine maleate and 60 mg of pseudoephedrine hydrochloride. Copyright © 2004 John Wiley & Sons, Ltd.
The Hummel–Dreyer method: impact in pharmacology
Authors: Šoltés Ladislav
Published: May 2004

Reviews devoted to drug–protein binding present mostly a variety of analytical methods applied to studying the unique process when the protein reversibly binds a low‐molecular‐weight substance—the drug. This report, however, reviews the great majority of papers presenting the application of one particular method introduced by Hummel and Dreyer in 1962: Measurement of protein‐binding phenomena by gel ﬁltration. The main criterion in reviewing published papers was the application of the Hummel–Dreyer method when the counterpart substances were the drug and the human plasma protein. Copyright © 2003 John Wiley & Sons, Ltd.
Investigation into the use of C‐ and N‐terminal GFP fusion proteins for subcellular localization studies using reverse transfection microarrays
Authors: Palmer Ella , Freeman Tom
Published: May 2004

Reverse transfection microarrays were described recently as a high throughput method for studying gene function. We have investigated the use of this technology for determining the subcellular localization of proteins. Genes encoding 16 proteins with a variety of functions were placed in Gateway expression constructs with 3′ or 5′ green fluorescent protein (GFP) tags. These were then packaged in transfection reagent and spotted robotically onto a glass slide to form a reverse transfection array. HEK293T cells were grown over the surface of the array until confluent and GFP fluorescence visualized by confocal microscopy. All C‐terminal fusion proteins localized to cellular compartments in accordance with previous studies and/or bioinformatic predictions. However, less than half of the N‐terminal fusion proteins localized correctly. Of those that were not in concordance with the C‐terminal tagged proteins, half did not exhibit expression and the remainder had differing subcellular localizations to the C‐terminal fusion protein. This data indicates that N‐terminal tagging with GFP adversely affects the protein localization in reverse transfection assays, whereas tagging with GFP at the C‐terminal is generally better in preserving the localization of the native protein. We discuss these results in the context of developing high‐throughput subcellular localization assays based on the reverse transfection array technology. Copyright © 2004 John Wiley & Sons, Ltd.
Mapping the Gene Ontology into the Unified Medical Language System
Authors: Lomax Jane , McCray Alexa T.
Published: May 2004

We have recently mapped the Gene Ontology (GO), developed by the Gene Ontology Consortium, into the National Library of Medicine's Unified Medical Language System (UMLS). GO has been developed for the purpose of annotating gene products in genome databases, and the UMLS has been developed as a framework for integrating large numbers of disparate terminologies, primarily for the purpose of providing better access to biomedical information sources. The mapping of GO to UMLS highlighted issues in both terminology systems. After some initial explorations and discussions between the UMLS and GO teams, the GO was integrated with the UMLS. Overall, a total of 23% of the GO terms either matched directly (3%) or linked (20%) to existing UMLS concepts. All GO terms now have a corresponding, official UMLS concept, and the entire vocabulary is available through the web‐based UMLS Knowledge Source Server. The mapping of the Gene Ontology, with its focus on structures, processes and functions at the molecular level, to the existing broad coverage UMLS should contribute to linking the language and practices of clinical medicine to the language and practices of genomics. Copyright © 2004 John Wiley & Sons, Ltd.
Design, implementation and maintenance of a model organism database for Arabidopsis thaliana
Authors: Weems Danforth , Miller Neil , Garcia‐Hernandez Margarita , Huala Eva , Rhee Seung Y.
Published: May 2004

The Arabidopsis Information Resource (TAIR) is a web‐based community database for the model plant Arabidopsis thaliana. It provides an integrated view of genes, sequences, proteins, germplasms, clones, metabolic pathways, gene expression, ecotypes, polymorphisms, publications, maps and community information. TAIR is developed and maintained by collaboration between software developers and biologists. Biologists provide specification and use cases for the system, acquire, analyse and curate data, interact with users and test the software. Software developers design, implement and test the database and software. In this review, we briefly describe how TAIR was built and is being maintained. Copyright © 2004 John Wiley & Sons, Ltd.
Comparative genomic assessment of novel broad‐spectrum targets for antibacterial drugs
Authors: White Thomas A. , Kell Douglas B.
Published: May 2004

Single and multiple resistance to antibacterial drugs currently in use is spreading, since they act against only a very small number of molecular targets; finding novel targets for anti‐infectives is therefore of great importance. All protein sequences from three pathogens (Staphylococcus aureus, Mycobacterium tuberculosis and Escherichia coli O157:H7 EDL993) were assessed via comparative genomics methods for their suitability as antibacterial targets according to a number of criteria, including the essentiality of the protein, its level of sequence conservation, and its distribution in pathogens, bacteria and eukaryotes (especially humans). Each protein was scored and ranked based on weighted variants of these criteria in order to prioritize proteins as potential novel broad‐spectrum targets for antibacterial drugs. A number of proteins proved to score highly in all three species and were robust to variations in the scoring system used. Sensitivity analysis indicated the quantitative contribution of each metric to the overall score. After further analysis of these targets, tRNA methyltransferase (trmD) and translation initiation factor IF‐1 (infA) emerged as potential and novel antimicrobial targets very worthy of further investigation. The scoring strategy used might be of value in other areas of post‐genomic drug discovery. Copyright © 2004 John Wiley & Sons, Ltd.
Chiral investigation of midodrine, a long‐acting α‐adrenergic stimulating agent
Authors: Quaglia M.G. , Farina A. , Palmery M. , Desideri N. , Donati E. , Bossù E. , Strano S.
Published: May 2004

Midodrine hydrochloride is a peripheral α1‐adrenoreceptor agonist that induces venous and arterial vasoconstriction. Midodrine, after oral or intravenous administration, undergoes enzymatic hydrolysis and releases deglymidodrine, a pharmacologically active metabolite. Midodrine and deglymidodrine have a chiral carbon in the 2‐position. To investigate the bioactivity of racemates and enantiomers of the drug and metabolite, three chromatographic chiral stationary phases, Chiralcel OD‐H, Chiralcel OD‐R, and α1‐AGP, were evaluated for enantiomeric resolution. Good enantioseparation of midodrine racemate was obtained using the Chiralcel OD‐H column. This stationary phase was then used to collect separately the midodrine enantiomers. By alkaline hydrolysis of rac‐midodrine and each separated enantiomer, rac‐deglymidodrine and its enantiomers were prepared. The control of the enantiomeric purity was carried out by α1‐AGP stationary phase, while the hydrolysis of rac‐midodrine and its enantiomers was controlled by capillary electrophoresis using trimethyl‐β‐cyclodextrin as chiral selector. The pharmacological activity of the two racemates and the two enantiomeric pairs was tested in vitro on a strip of rabbit descending thoracic aorta. The tests continued that the activity of the drug and metabolite is due only to the (−)‐enantiomer because neither of the (+)‐enantiomers is active. Chirality 16:356–362, 2004. ©2004 Wiley‐Liss, Inc.
Wnt genes define distinct boundaries in the developing human brain: Implications for human forebrain patterning
Authors: Abu‐Khalil A. , Fu L. , Grove E.A. , Zecevic N. , Geschwind Daniel H.
Published: May 2004

Understanding the factors that govern human forebrain regionalization along the dorsal–ventral and left–right (L‐R) axes is likely to be relevant to a wide variety of neurodevelopmental and neuropsychiatric conditions. Recent work in lower vertebrates has identified several critical signaling molecules involved in embryonic patterning along these axes. Among these are the Wingless‐Int (WNT) proteins, involved in the formation of dorsal central nervous system (CNS) structures, as well as in visceral L‐R asymmetry. We examined the expression of WNT2b and WNT7b in the human brain, because these genes have highly distinctive expression patterns in the embryonic mouse forebrain. In the human fetal telencephalon, WNT2b expression appears to define the cortical hem, a dorsal signaling center previously characterized in mouse, which is also confirmed by BMP7 expression. In diencephalon, WNT2b expression is restricted to medial dorsal structures, including the developing pineal gland and habenular nucleus, both implicated in CNS L‐R asymmetry in lower organisms. At 5 weeks gestation, WNT7b is expressed in cerebral cortical and diencephalic progenitor cells. As the cortical plate develops, WNT7b expression shifts, demarcating deep layer neurons of the neocortex and the hippocampal formation. Spatial and temporal expression patterns show startling similarity between human and mouse, suggesting that the developmental roles of these WNT genes may be highly conserved, despite the far greater size and complexity of the human forebrain. J. Comp. Neurol. 474:276–288, 2004. © 2004 Wiley‐Liss, Inc.
Paucity of horizontal connections for binocular vision in V1 of naturally strabismic macaques: Cytochrome oxidase compartment specificity
Authors: Tychsen Lawrence , Ming‐Fong Wong Agnes , Burkhalter Andreas
Published: May 2004

To describe the structural basis for lack of binocular fusion in strabismic primates, we investigated intrinsic horizontal connections within striate cortex (area V1) of normal and strabismic, adult macaque monkeys. The strabismic animals had early‐onset natural esotropia (the visual axes deviated nasally), normal visual acuity in each eye, and the constellation of ocular motor deficits that typify human infantile strabismus. Horizontal patchy connections and synaptic boutons were labeled by injections of the neuronal tracer biotinylated dextran amine. Ocular dominance columns (ODCs), and blob vs. interblob compartments, were revealed by using cytochrome oxidase (CO). In layers 2/3 and 4B of the strabismic monkeys, patchy projections and boutons terminated much more frequently in same‐eye (73%) as opposed to opposite‐eye (27%) ODCs (normal monkeys 58% and 42%, respectively). The deficiency of binocular connections in the strabismic cortex was evident qualitatively as a “skip” pattern, in which every other row of ODCs had labeled patches. Analysis of V1 in normal monkeys revealed that the deficits in strabismic V1 were due mainly to a loss of binocular connections between neurons in CO‐interblob compartments. In both normal and strabismic monkeys: (1) CO‐blob compartment neurons showed a more pronounced bias for monocular connectivity, and (2) commitment of connections to the same CO‐compartment as the injection site (blob‐to‐blob, or interblob‐to‐interblob) was moderately strong (64%) but far from absolute. These findings help elucidate the relative roles of visual experience vs. innate mechanisms in the development of axonal connections between ocular dominance domains and compartments within macaque V1. They also provide the first detailed description of the V1 maldevelopments associated with unrepaired natural, infantile‐onset strabismus in primates. J. Comp. Neurol. 474:261–275, 2004. © 2004 Wiley‐Liss, Inc.
Cerebral areas mediating visual redirection of gaze: Cooling deactivation of 15 loci in the cat
Authors: Lomber Stephen G. , Payne Bertram R.
Published: May 2004

In humans, damage to posterior parietal or frontal cortices often induces a severe impairment of the ability to redirect gaze to visual targets introduced into the contralateral field. In cats, unilateral deactivation of the posterior middle suprasylvian (pMS) sulcus in the posterior inferior parietal region also results in an equally severe impairment of visually mediated redirection of gaze. In this study we tested the contributions of the pMS cortex and 14 other cortical regions in mediating redirection of gaze to visual targets in 31 adult cats. Unilateral cooling deactivation of three adjacent regions along the posterior bend of the suprasylvian sulcus (posterior middle suprasylvian sulcus, posterior suprasylvian sulcus, and dorsal posterior ectosylvian gyrus at the confluence of the occipital, parietal, and temporal cortices) eliminated visually mediated redirection of gaze towards stimuli introduced into the contralateral hemifield, while the redirection of gaze toward the ipsilateral hemifield remained highly proficient. Additional cortical loci critical for visually mediated redirection of gaze include the anterior suprasylvian gyrus (lateral area 5, anterior inferior parietal cortex) and medial area 6 in the frontal region. Cooling deactivation of: 1) dorsal or 2) ventral posterior suprasylvian gyrus; 3) ventral posterior ectosylvian gyrus, 4) middle ectosylvian gyrus; 5) anterior or 6) posterior middle suprasylvian gyrus (area 7); 7) anterior middle suprasylvian sulcus; 8) medial area 5; 9) the visual portion of the anterior ectosylvian sulcus (AES); 10) or lateral area 6 were all without impact on the ability to redirect gaze. In summary, we identified a prominent field of cortex at the junction of the temporo‐occipito‐parietal cortices (regions pMS, dPE, PS), an anterior inferior parietal field (region 5L), and a frontal field (region 6M) that all contribute critically to the ability to redirect gaze to novel stimuli introduced into the visual field during fixation. These loci have several features in common with cortical fields in monkey and human brains that contribute to the visually guided redirection of the head and eyes. J. Comp. Neurol. 474:190–208, 2004. © 2004 Wiley‐Liss, Inc.
Timing and topography of cell genesis in the rat retina
Authors: Rapaport David H. , Wong Lily L. , Wood Eric D. , Yasumura Douglas , LaVail Matthew M.
Published: May 2004

To understand the mechanisms of cell fate determination in the vertebrate retina, the time course of the generation of the major cell types needs to be established. This will help define and interpret patterns of gene expression, waves of differentiation, timing and extent of competence, and many of the other developmental processes involved in fate acquisition. A thorough retinal cell “birthdating” study has not been performed for the laboratory rat, even though it is the species of choice for many contemporary developmental studies of the vertebrate retina. We investigated the timing and spatial pattern of cell genesis using 3H‐thymidine (3H‐TdR). A single injection of 3H‐TdR was administered to pregnant rats or rat pups between embryonic day (E) 8 and postnatal day (P) 13. The offspring of prenatally injected rats were delivered and all animals survived to maturity. Labeled cells were visualized by autoradiography of retinal sections. Rat retinal cell genesis commenced around E10, 50% of cells were born by approximately P1, and retinogenesis was complete near P12. The first postmitotic cells were found in the retinal ganglion cell layer and were 9–15 μm in diameter. This range includes small to medium diameter retinal ganglion cells and large displaced amacrine cells. The sequence of cell genesis was established by determining the age at which 5, 50, and 95% of the total population of cells of each phenotype became postmitotic. With few exceptions, the cell types reached these developmental landmarks in the following order: retinal ganglion cells, horizontal cells, cones, amacrine cells, rods, bipolar cells, and Müller glia. For each type, the first cells generated were located in the central retina and the last cells in the peripheral retina. Within the sequence of cell genesis, two or three phases could be detected based on differences in timing, kinetics, and topographic gradients of cell production. Our results show that retinal cells in the rat are generated in a sequence similar to that of the primate retina, in which retinogenesis spans more than 100 days. To the extent that sequences reflect underlying mechanisms of cell fate determination, they appear to be conserved. J. Comp. Neurol. 474:304–324, 2004. © 2004 Wiley‐Liss, Inc.
Characterization of long‐term mouse brain aggregating cultures: Evidence for maintenance of neural precursor cells
Authors: Berglund C. Mikaela D. , Aarum Johan , Budd Haeberlein Samantha L. , Nyengaard Jens R. , Hökfelt Tomas , Sandberg Kristian , Näslund Jan , Persson Mats A.A.
Published: May 2004

An extensive characterization of fetal mouse brain cell aggregates has been performed using immunohistochemical and stereological methods. Single cell suspensions from mechanically dissociated cortex and hippocampus were cultured in serum‐free, B27‐supplemented medium under constant gyratory agitation for up to 56 days. Three‐dimensional aggregates started to form immediately after seeding and reached a final average size of 500 μm in diameter. Among the cell types identified, neurons were the most abundant cells in the aggregates, followed by astrocytes, microglia, and oligodendrocytes. Western blotting for synaptophysin and immunostaining for neurotransmitter‐related molecules indicated the presence of well‐defined phenotypic characteristics of the neurons in this culture system, suggesting functionality. Proliferating cells, many with neural precursor cell properties, were seen throughout the culture period and could be isolated from the aggregates even after 2 months in culture. Neural precursor cells were isolated from the aggregates after more than 1 month in culture; these cells were successfully differentiated into neurons, astrocytes, and oligodendrocytes. The aggregate culture system may provide a versatile tool for molecular dissection of processes identified in mouse models, including transgenic animals and manipulation of neural precursor cells. J. Comp. Neurol. 474:246–260, 2004. © 2004 Wiley‐Liss, Inc.
Obtaining high rates for the etching and ashing for large liquid‐crystal display substrates by using a surface‐wave plasma source
Authors: Aoki Katsuaki , Uekusa Shin‐ichiro , Yamauchi Takeshi , Yamage Masashi , Saito Shuichi , Takagi Shigeyuki , Kanoh Masaaki
Published: May 2004

In the manufacture of thin‐film transistor liquid‐crystal displays (TFT‐LCDs), a major goal is to scale up the production of liquid‐crystal panels and lower their production costs. For that reason, we have developed a large‐area high‐density (3.58 × 1011 cm−3) surface‐wave plasma source based on a slot antenna. This source is designed to reduce the number of separate production steps in semiconductor processing, thereby increasing the throughput of large‐area TFT‐LCD substrates. We are convinced that the use of this plasma source will not only increase the etching rate, but also reduce the need for industry‐standard photoresist liftoff processes by enabling an uninterrupted transition after etching to photoresist liftoff (ashing). We found that the etching rate for a 400 × 500 mm2 polysilicon layer on a glass substrate was 58 nm/min with a uniformity of ±6.8% when our source was used. Furthermore, when ashing was carried out in the same chamber immediately after etching, the ashing rate was 1430 nm/min with a uniformity of ±9.3%. We also found that the selectivity ratio for resist versus polysilicon and the SiO2 in an underlayer was higher than 140. This implies that our process outperforms the standard resist liftoff process; indeed, we have realized a high‐uniformity etching process with twice the etching rate of standard methods. In addition, we have made progress toward realizing an ashing process, which is almost ready for practical use. These two processes can be combined into a single integrated etching–ashing process that will reduce the need for a separate liftoff procedure and simplify processing. © 2004 Wiley Periodicals, Inc. Electron Comm Jpn Pt 2, 87(6): 28–38, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjb.10086
Optical observation technique for SH‐type surface acoustic wave
Authors: Miyamoto Akinori , Matsuda Satoru , Wakana Shinichi , Ito Akio
Published: May 2004

A new technology has been developed for optically visualizing the intensity distribution of the acoustic wave within a surface acoustic wave (SAW) device. In this technique, the strain generated by the SH (Shear Horizontal) wave in a LiTaO3crystal is detected via the photoelastic effect as a change of polarization state of the light passing through the device. By detecting the SH component, which is the main component of the SAW on a 42°YX LiTaO3, the method provides for more direct observation of the behavior of the acoustic wave in a device than in the conventional technique that detects the SV (Shear Vertical) component, a secondary component. Based on the present technique, an acoustic wave observation setup is developed and a simple resonator is evaluated. It is found that the acoustic‐wave distributions within an IDT depend on the driving frequencies and that acoustic wave energy is leaked outside the resonator at specific frequencies. Further, a more accurate visualization of the acoustic wave component is made possible by correcting for the influence of the electro‐optical effect on the observation results. Another setup has been developed in which a probe light beam is incident from the back side of the substrate and the light reflected from the device surface is used in order to determine the distribution under an electrode. © 2004 Wiley Periodicals, Inc. Electron Comm Jpn Pt 2, 87(6): 10–17, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjb.20066
Rapid environment adaptation method based on HMM composition with prior noise GMM and multi‐SNR models for noisy speech recognition
Authors: Ida Masaki , Nakamura Satoshi
Published: May 2004

In the use of speech recognition systems in a real environment, it is inevitable that surrounding environmental noise is present in the input speech, which degrades recognition performance. It is difficult in most cases to predict the mixing of the noise, and the discrepancy of noise environments between the input signal and the acoustic model is a reason for degradation of recognition performance. Consequently, it is desirable to construct an acoustic model which is robust to the mixing of various kinds of noise. The problem of noise mixture can be divided into two aspects, namely, diversified kinds of noise and diversified values of the SNR. In this paper, HMM composition using weight adaptation of the noise GMM is applied to the first problem, and the multi‐SNR path model is applied to the second problem. Performance evaluation is performed for a combination of these two approaches in a speech recognition experiment in a noisy environment, using the travel conversation task and the AURORA2 task. When 1 second of adaptation data is used in the AURORA2 task for SNR = 5 dB, the recognition rate is improved by 53% compared to the baseline HMM. This corresponds to the case in which 10 seconds of adaptation data is used in conventional HMM composition. © 2004 Wiley Periodicals, Inc. Electron Comm Jpn Pt 2, 87(6): 39–48, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjb.20093
Road extraction VLSI processor based on optimal allocation and its application to highly safe intelligent vehicles
Authors: Hariyama Masanori , Kudoh Takao , Kameyama Michitaka
Published: May 2004

This paper proposes a collision warning system that gives a warning to a driver by detecting a hazard during driving. This system has a configuration based on the fail‐safe concept that gives a warning to the driver by determining the state of danger when a path that the vehicle cannot safely drive on cannot be found within a certain processing time. Furthermore, it proposes a road extraction VLSI processor for a highly safe vehicle based on the logic‐in‐memory architecture, which eliminates transfer bottlenecks by integrating the memory and the functional units. This VLSI processor uses a VLSI‐oriented algorithm based on regular repetitions of local parallel processing of external three‐dimensional coordinate information. A memory system that allows parallel accessing with a minimal amount of hardware is desirable in designing a VLSI processor. For this purpose, optimal memory allocation for parallel accessing designed for minimal memory capacity is important. In order to resolve the problem of increased search space for obtaining optimal allocation in global search, a method of limiting the search space by focusing on the periodicity of the three‐dimensional coordinates of the memory module and the data stored in it is proposed. Evaluation of this VLSI processor reveals a significant reduction of the chip area under identical performance conditions. © 2004 Wiley Periodicals, Inc. Electron Comm Jpn Pt 2, 87(6): 49–57, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjb.20094