bluuebunny/crossref_metadata_2025 huggingface dataset:
Labelling of anilines, benzylamines and some N‐heterocyclics using cycloocta‐1,5‐dienyliridium(I)‐1,1,1,5,5,5‐hexafluoro‐pentan‐2,4‐dionate and isotopic hydrogen gas in DMF or DMA
Authors: Garman R. N. , Hickey M. J. , Kingston L. P. , McAuley B. , Jones J. R. , Lockley W. J. S. , Mather A. N. , Wilkinson D. J.
Published: November 2004

A wide range of anilines, benzylamines and some N‐heterocyclics can be ortho‐deuterated at room temperature using deuterium gas and cycloocta‐1,5‐dienyliridium(I)‐1,1,1,5,5,5‐hexafluoropentan‐2,4‐dionate in DMF or DMA. The method is applicable to labelling with tritium. Copyright © 2004 John Wiley & Sons, Ltd.
Downregulation of complexin I and complexin II in the medial thalamus is blocked by N‐acetylcysteine in experimental Wernicke's encephalopathy
Authors: Hazell Alan S. , Wang Chunlei
Published: November 2004

Metabolic dysfunction as a consequence of thiamine (vitamin B1) deficiency (TD), a model of Wernicke's encephalopathy, leads to elevation of extracellular glutamate concentration in vulnerable brain regions consistent with the development of excitotoxicity. Complexin I and complexin II are two genes labeling principally inhibitory and excitatory synapses, respectively. Because current evidence supports an important role for complexins in the modulation of neurotransmitter release, we examined the involvement of both proteins in the pathology of the medial thalamus and inferior colliculus in TD rats by immunoblotting. At the symptomatic stage, complexin I and complexin II levels in the medial thalamus were decreased by 63% and 45%, respectively, compared to control animals, but were unchanged in the inferior colliculus. These changes in thalamus were also observed using immunohistochemical methods, and seemed to be due to downregulation of both proteins because synaptophysin levels were unaffected in this brain region. In addition, cotreatment with the antioxidant N‐ acetylcysteine prevented both neuronal loss and downregulation of complexins. Our findings suggest dysregulation of excitatory and inhibitory neurotransmitter release in the medial thalamus, which is not present in the inferior colliculus. Furthermore, loss of complexin I and II in the thalamus may be mediated by processes that involve oxidative stress. Such changes in complexin levels may contribute to the pathophysiology of thalamic damage in TD, and offer a potential basis for the well‐known differences in pathology between this structure and the inferior colliculus in this disorder. © 2004 Wiley‐Liss, Inc.
The travel cost method and the economic value of leisure time
Authors: Douglas Aaron J. , Johnson Richard L.
Published: November 2004

Recent estimates of high values for tourist related recreation USA amenity values indicate that allocation of basic water and terrestrial resources to recreation activities should be given precedence over conventional market oriented activities that often degrade or even extirpate the resource. We discuss at length the travel cost method (TCM), a survey based technique that quantifies the non‐market benefits of trips to recreation sites. The TCM has been cast in the role of an ‘umpire’ in recent resource allocation debates. Understanding the key role of the TCM in the debate will aid tourist agency officials throughout the world. This article is a U.S. Government work and is in the public domain in the U.S.A.
Comparison between Polymorphic Behaviors of Ziegler‐Natta and Metallocene‐Made Isotactic Polypropylene: The Role of the Chain Microstructure
Authors: De Rosa Claudio , Auriemma Finizia , Spera Clementina
Published: November 2004

An analysis of the polymorphic behavior of samples of isotactic polypropylene (iPP) prepared with heterogeneous Ziegler‐Natta catalysts is presented. A comparison with the crystallization properties of iPP samples prepared with a single center homogeneous metallocene catalyst is also shown. Samples of Ziegler‐Natta iPP samples, prepared with MgCl2‐supported catalysts, have been fractionated by extraction with boiling solvents. The irregular fraction, insoluble in diethyl ether and soluble in hexane, crystallizes from the melt almost totally in the γ form. The more stereoregular fractions crystallize instead basically in the α form. The relative amount of γ form crystallized from the melt is much lower that that observed in samples of metallocene‐made iPP containing comparable amount of defects. While in the metallocene‐made iPP the distribution of defects along the chains is random, in Ziegler‐Natta iPP samples the majority of defects are segregated in a small fraction of poorly crystallizable macromolecules or in more irregular portions of the chain, so that much longer fully isotactic sequences can be produced, leading to the crystallization of the α form. This analysis allows concluding that some fractions of Ziegler‐Natta iPP are characterized by chains with a stereo‐block microstructure, consisting in regular isotactic sequences linked to more irregular sequences.
Micro‐Rheological Modeling of Flow‐Induced Crystallization in Mixed Shear/Extensional Flows
Authors: Coppola Salvatore , Grizzuti Nino
Published: November 2004

Flow Induced Crystallization (FIC) is the common term to indicate the acceleration in polymer crystallization kinetics due to the action of flow. When modeling FIC, two major challenges are encountered. On the one hand, the model must be able to produce quantitative reliable results, while correctly describing the coupling between the intrinsic (quiescent) crystallization kinetics and the rheological response of the polymer. On the other hand, the model must be able to describe the complex kinematics taking place in real industrial processes. In this paper, we present the predictions of a recently proposed model for FIC in the case of a mixed flow, where both shear and extensional components are present at the same time. In particular, the effects of the overall flow intensity and of relative weight between shear and extension on the enhancement in nucleation rate are presented and discussed. Some guidelines for future development are also proposed.
Blends Based on Polystyrene and Statistical Aromatic Copolycarbonates
Authors: Penco Maurizio , Sartore Luciana , Bigotti Fabio , Spagnoli Gloria , Ferrari Fabrizio , D'Antone Salvatore
Published: November 2004

The compatibility of polystyrene (PS) with aromatic copolycarbonates containing bisphenol A (BPA) and tetramethyl bisphenol A (TMBPA) was investigated. The simple prevision scheme developed by Sonja Krause was employed to evaluate the effect of the copolymer molecular structure on the miscibility with polystyrene. These prevision data were used to select copolycarbonates of potential interest. Statistical copolycarbonates (CPC) containing different BPA/TMBPA molar ratios were synthesised by polycondensation reaction between a mixture of the two monomers and phosgene. PS/polycarbonates blends, prepared by casting from chloroform solution, were studied with differential scanning calorimetry and optical microscopy to evaluate the components compatibility, which increases with the TMBPA copolymer content.
Engineered Green Fluorescence Proteins for Proteomics and Biomolecular Electronic Applications
Authors: Bizzarri Ranieri , Pellegrini Vittorio , Arcangeli Caterina , Ferrari Aldo , Nifosì Riccardo , Pingue Pasqualantonio , Tozzini Valentina , Giacca Mauro , Beltram Fabio
Published: November 2004

The discovery of the photochromic characteristics of engineered green fluorescent proteins (GFPs) allows new proteomics and biomolecular electronic applications. In particular, photoreversibility among two distinct optical states can lead to the realization of a bio‐optical high density storage memory. Here we review our recent work on an optically bistable GFP and we report the recent developments of self‐assembly methods for spatial immobilization of proteins into well‐definite 2D patterns.
(4‐Vinylpyridine‐Styrene) Copolymer as Host Polymer for Chromophoric Complexes with Potential Second Order Nonlinear Optical Properties
Authors: Borbone Fabio , Caruso Ugo , De Maria Antonella , Fusco Michela , Panunzi Barbara , Roviello Antonio
Published: November 2004

A series of new metal containing polymers for second order nonlinear optics have been prepared by grafting Cu (II) and Pd (II) chromophoric complexes on a preformed (4‐vinylpyridine‐styrene) copolymeric backbone. The metallated polymers have been chemically and physically characterized. They show high glass transition temperatures, high thermal stability and good solubility. Their properties have been compared with analogous metallated poly(4‐vinylpyridine) samples: variations in the polymeric backbone, as well as in ligands, metal, and metallation ratio, allow to tune their properties.
On solution of Lamé equations in axisymmetric domains with conical points
Authors: Nkemzi Boniface
Published: November 2004

Partial Fourier series expansion is applied to the Dirichlet problem for the Lamé equations in axisymmetric domains $\hat{\Omega}$⊂ℝ3 with conical points on the rotation axis. This leads to dimension reduction of the three‐dimensional boundary value problem resulting to an infinite sequence of two‐dimensional boundary value problems on the plane meridian domain Ωa⊂ℝ of $\hat{\Omega}$ with solutions un(n=0,1,2,…) being the Fourier coefficients of the solution û of the 3D BVP. The asymptotic behaviour of the Fourier coefficients un (n=0,1,2,…) near the angular points of the meridian domain Ωa is fully described by singular vector‐functions which are related to the zeros αn of some transcendental equations involving Legendre functions of the first kind. Equations which determine the values of αn are given and a numerical algorithm for the computation of αn is proposed with some plots of values obtained presented. The singular vector functions for the solution of the 3D BVP is obtained by Fourier synthesis. Copyright © 2004 John Wiley & Sons, Ltd
Intracellular Ca2+ regulation in a human prostate stromal cell culture
Authors: Wu C , Fry PM , Sui G , Fry CH
Published: November 2004

Aims

Prostate stromal cell cultures are used in vitro to study the cellular pathophysiology of benign prostatic hyperplasia (BPH), but their functional properties are poorly understood. This study characterized intracellular Ca2+ ([Ca2+]i) regulation in a cultured cell line in comparison to freshly isolated cells, as a background to understanding contractile regulation and cellular proliferation in this tissue.

Methods

Prostate stromal cells were isolated from either PrS6 cell cultures, with an extended life span by transfection with the SV40 T‐antigen, tsA58‐U19, or freshly obtained transition zone prostate samples, primary cells. [Ca2+]i was measured in vitro with the indicator Fura‐2 by epifluorescence microscopy.

Results

Phenylephrine, high‐K+, and caffeine induced Ca2+‐transients in primary cells (resting [Ca2+]i 94 ± 8 nM, n = 29; peak 193 ± 26 nM, n = 19). In PrS6 cells resting [Ca2+]i was 96 ± 8 nM (n = 78) and in 34 of these 78 cells, 30 μM phenylephrine increased [Ca2+]i to 296 ± 28 nM. 5‐methyl‐urapidil (10–30 μM) inhibited this response in 10 of 16 cells. Spontaneous Ca2+‐transients were also observed in 91% of phenylephrine‐responsive cells, but in only 20% of non‐responsive cells (P < 0.01). Ca2+‐transients were also induced by high‐K+ solution, and 20 mM caffeine. The latter abolished the response to subsequent phenylephrine application. Depletion of intracellular Ca2+ stores by caffeine or restoration from a Ca2+‐free superfusate caused a substantial rise of [Ca2+]i.

Conclusions

PrS6 prostate stromal cells express functional α1‐adrenoceptors associated with spontaneous intracellular Ca2+‐transients. They exhibit functional Ca2+ channels, intracellular Ca2+ stores, and Ca2+ entry induced by store depletion. Stromal cultures can therefore be used to characterize the cellular physiology of prostate stromal cell contraction and proliferation. © 2004 Wiley‐Liss, Inc.
On Dimensional Adaptivity For Mixed Beam‐Shell‐Structures
Authors: Wenzel Michael
Published: December 2004

A hierarchical model for dimensional adaptivity, using mixed beam‐shell structures, is presented. Thin‐walled beam structures are often calculated on the base of beam theories. Parts of the global structure, like framework corners, are usually analyzed with shell elements in a separate model. To minimize the modeling and calculation expense, a transition element to couple beam and shell structures is used. A dimensional adaptiv algorithm is introduced to automate this the procedure of modeling and calculation. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
The research on physiological property of functionalized hyaluronan: interaction between sulfated hyaluronan and plasma proteins
Authors: Satoh Takashi , Nishiyama Koh , Nagahata Misao , Teramoto Akira , Abe Koji
Published: November 2004

Hyaluronan (HA) is a natural polysaccharides which has no sulfated group but high molecular weight in comparison with other glycosaminoglycans (GAGs). Previously it has been cleared up that the cell function of human keratinocytes is affected by S‐HA (HA substituted with sulfated groups). Most biomedical materials contact with blood components, proteins, cells, etc. In this study, the interaction between S‐HA and blood components is discussed, that is, plasma proteins. And the application of S‐HA for new analytical and separation method of some proteins is pointed out. None of the proteins were adsorbed to HA. Fibronectin and fibinogen were adsorbed to S‐HA, but immunoglobulin‐G and insulin were not adsorbed to it as well as heparin. However, albumin could interact only with heparin, and it did not interact with S‐HA. Furthermore S‐HA adsorbed the plasma proteins that did not adsorb to heparin. It is clear that S‐HA showed different interaction with the plasma proteins in comparison with natural sulfated polysaccharides. Copyright © 2004 John Wiley & Sons, Ltd.
Structure and properties of CE/CTBN/EP blends: II. Effect of EP on the mechanical properties and thermostability of the CE/CTBN system
Authors: Feng Yu , Fang Zhengping , Gu Aijuan
Published: December 2004

Carboxyl‐terminated butadiene‐acrylonitrile rubber (CTBN) has often been used to improve the toughness of cyanate ester (CE) resin while sacrificing modulus and thermostability. In this paper, the addition of the appropriate amount of epoxy resin (EP) to the CE/CTBN system is shown to not only increase the modulus and thermostability of the blend, but also improve the toughness. The values of impact strength showed a maximum for the CE/CTBN/EP 100/5/5 blend. The temperature of 10 % weight loss (T10) improves from 376 °C for CE/CTBN 100/5 to 407 °C for the CE/CTBN/EP 100/5/2.5 blend. It is proposed that addition of the appropriate amount of EP can decrease the mobility and increase the stability of CTBN via the reaction between the terminal carboxyl group of CTBN and the hydroxyl group of EP. But a very high EP concentration will decrease the crosslinking density of CE, consequently reducing the mechanical properties and thermostability of the blends. Copyright © 2004 Society of Chemical Industry
Phosphonation of polysulfones via lithiation and reaction with chlorophosphonic acid esters
Authors: Lafitte Benoît , Jannasch Patric
Published: November 2004

A noncatalytic route for the phosphonation of polysulfones was established in which lithiated sites on polysulfones were reacted with an excess of chlorophosphonic acid esters through an SNP(V) mechanism. Both the bisphenol A and biphenyl sulfone segments of the polysulfone main chain were modified according to whether brominated polysulfone or pristine polysulfone was used. Up to 50% of the repeating units of the polysulfones were modified by a careful selection of reaction parameters to avoid crosslinking. The phosphonated polysulfones in their acid form showed high thermal stability with decomposition temperatures of approximately 350 °C under nitrogen. Polysulfones with phosphonated bisphenol A segments showed good membrane‐forming properties and are candidates for components in ionomer composite membranes for fuel cells. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 273–286, 2005
Cryogenic sediment‐filled wedges, northern Delaware, USA
Authors: Lemcke Mary D. , Nelson Frederick E.
Published: November 2004

Wedge‐like sedimentary structures at two sites in northern Delaware USA are located at the erosional surface of, and extend into, channel deposits of the fluvial, mid‐Pleistocene Columbia Formation. The wedges are 0.25–0.60 m wide at the top, 1.0–1.5 m in vertical extent, contain moderate to poorly sorted, and vertically‐stratified sediment, and are overlain unconformably by a layer of wind‐blown silt. Several hypotheses for the formation and infill of the wedges were evaluated using detailed physical, stratigraphic, and sedimentological information. The most likely explanation for the features is that they are relict cryogenic structures formed by thermal‐contraction cracking in permafrost, and filled with wind‐blown sediments derived from the Columbia Formation. The wedges are believed to have formed in the tundra environment that existed in northern Delaware, south of the Laurentide Ice Sheet, during the coldest parts of the Wisconsinan glaciation. Copyright © 2004 John Wiley & Sons, Ltd.
CFTR: More than just a chloride channel
Authors: Mehta Anil
Published: November 2004

This review examines the cystic fibrosis transmembrane conductance regulator (CFTR) protein. After summarizing the ion channels regulated by CFTR, the review focuses on the functions of CFTR that do not relate directly to a disease mechanism based on a channelopathy. The key concept is that newly synthesized CFTR has to enter lipid vesicles which bud from the endoplasmic reticulum. This is abnormally low in ΔF508 CFTR. Normal wild type vesicular CFTR enters a recycling pool of lipid vesicles which transiently dock with the apical membrane only for CFTR to be retrieved shortly after into a sub‐apical recycling compartment. This retrieval is abnormally fast in ΔF508 CFTR. The review discusses the relationship between this process and the difficult topic of fat metabolism and then explores the possible links between abnormal fatty acid turnover and inflammatory cascades that are abnormal in cystic fibrosis. Finally the review concentrates on the emerging functions of a protein kinase (AMP‐activated kinase) which is bound near the C terminus of the CFTR protein whose functions could intergrate some of the abnormalities in lipid metabolism that result from mislocalization of CFTR in clinical disease. Pediatr Pulmonol. 2005; 39:292–298. © 2004 Wiley‐Liss, Inc.
Predicting protein functional sites with phylogenetic motifs
Authors: La David , Sutch Brian , Livesay Dennis R.
Published: November 2004

In this report, we demonstrate that phylogenetic motifs, sequence regions conserving the overall familial phylogeny, represent a promising approach to protein functional site prediction. Across our structurally and functionally heterogeneous data set, phylogenetic motifs consistently correspond to functional sites defined by both surface loops and active site clefts. Additionally, the partially buried prosthetic group regions of cytochrome P450 and succinate dehydrogenase are identified as phylogenetic motifs. In nearly all instances, phylogenetic motifs are structurally clustered, despite little overall sequence proximity, around key functional site features. Based on calculated false‐positive expectations and standard motif identification methods, we show that phylogenetic motifs are generally conserved in sequence. This result implies that they can be considered motifs in the traditional sense as well. However, there are instances where phylogenetic motifs are not (overall) well conserved in sequence. This point is enticing, because it implies that phylogenetic motifs are able to identify key sequence regions that traditional motif‐based approaches would not. Further, phylogenetic motif results are also shown to be consistent with evolutionary trace results, and bootstrapping is used to demonstrate tree significance. Proteins 2005. © 2004 Wiley‐Liss, Inc.
Phenomenological description of polarization switching in ferroelectric semiconductors with charged defects
Authors: Morozovska Anna N. , Eliseev Eugene A.
Published: December 2004

We have proposed the phenomenological description of polarization switching peculiarities in ferroelectric semiconductors with charged defects and prevailing extrinsic conductivity. More precisely, we have modified the Landau–Ginsburg approach and shown that the macroscopic state of the aforementioned inhomogeneous system can be described by three coupled equations for three order parameters. Both the experimentally observed coercive field values, well below the thermodynamic one, and the various hysteresis loop deformations (minor, constricted and double loops) have been obtained in the framework of our model. The obtained results qualitatively explain the ferroelectric switching in such bulk ferroelectric materials as SBN single crystals doped with Ce, lead zirconate titanate (PZT) films doped with Nd, and La‐doped PZT ceramics. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Magnetooptics of quintet states of Tb3+ ion in terbium–yttrium aluminum garnet
Authors: Valiev U. V. , Gruber J. B. , Zandi B. , Rustamov U. R. , Rakhmatov A. S. , Dzhuraev D. R. , Narzullaev N. M.
Published: December 2004

The luminescence spectra and the degrees of magnetic circular polarization of luminescence (MCPL) have been studied in paramagnetic garnet Tb0.2Y2.8Al5O12 (YAG:Tb3+) for the radiative 4f–4f transition 5D4 ® 7F6. The features of the spectral dependencies of the MCPL degree with allowed (in symmetry) optical transitions between the Stark sublevels of the ground 7F6 and excited 5D4 multiplets of the non‐Kramers rare‐earth Tb3+ ion have been found in the YAG garnet structure. In the paramagnetic garnet Tb0.2Y2.8Al5O12 within the close ultraviolet spectral range, the non‐trivial features of the spectra of magnetic circular dichroism (MCD) have been found; they evidence a significant contribution of the quasi‐doublet states in the multiplets 5D3, 5G5 and 5L10 to magnetooptics of the 4f–4f transitions. The results of numerical calculations of the energy and wave functions have been presented for some Stark sublevels of the 5D4 and 7F6 multiplets of the Tb3+ ion in the crystalline field of D2 symmetry. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Phosphorus–hydrogen complexes in LEC‐grown InP
Authors: Ulrici W. , Kwasniewski A. , Czupalla M. , Neubert M.
Published: December 2004

In LEC‐grown InP, about 30 sharp vibrational absorption lines are measured in the frequency region 2200 to 2350 cm–1. All these lines are due to phosphorus‐hydrogen stretching modes. Experiments on InP containing both hydrogen and deuterium finally proved that the line at 2202.4 cm–1 is due to a single hydrogen atom bonded to P in an indium vacancy (VIn) and that the line at 2315.6 cm–1 is due to the complex of four P–H bonds in an VIn. In InP:H:D, this VInH4 complex gives rise to six vibrational lines in the region of P–H modes and six lines in the region of P–D modes because of the five different types of VInHnDm complexes. The measured frequencies of these 12 lines are in excellent agreement with those obtained from ab initio calculations reported in the literature. Additional P–H complexes are discussed. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Reply to “Comment on ‘Orbital ordering and orbital fluctuations in transition metal oxides’” [phys. stat. sol. (b) 242, No. 4, 962 (2005)]
Authors: Oleś Andrzej M.
Published: December 2004

We point out that the high degeneracy of 3T1g ground state of V3+ ions in LaVO3, used to derive the spin‐orbital model analyzed in the previous paper [A. M. Oleś, phys. stat. sol. (b) 236, 281 (2003)], allows for the orbital fluctuations which stabilize the ferromagnetic bonds in the C‐type antiferromagnetic phase of cubic vanadates. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Recent advances in the treatment of cystinosis
Authors: Schneider J. A. , Clark K. F. , Greene A. A. , Reisch J. S. , Markello T. C. , Gahl W. A. , Thoene J. G. , Noonan P. K. , Berry K. A.
Published: November 2004

Cysteamine bitartrate capsules (Cystagon) have been approved by the US Food and Drug Administration for use in patients with nephropathic cystinosis. Plasma cysteamine concentrations were virtually identical at various times following ingestion of either cysteamine hydrochloride or Cystagon capsules in 24 normal control subjects. A transfer study was done with eight cystinosis patients who had been receiving either cysteamine hydrochloride or phosphocysteamine for many years. The plasma cysteamine concentration was significantly higher 2h after Cystagon and the leukocyte cystine content was significantly lower at all times after Cystagon compared to older forms of the drug. These differences are probably the result of greater patient compliance in taking the capsules compared to the older, liquid forms of the drug. A new method for following the course of renal glomerular deterioration in diseases such as cystinosis has been published recently. This method was used to re‐analyse data on the efficacy of cysteamine treatment and to re‐analyse new data on treating cystinosis patients with either of two doses of cysteamine (1.30 g/m2 per day and 1.95 g/m2 per day). This new method agrees well with other methods and shows that both doses of drug are equally effective in maintaining glomerular function.
The hepatic glycogen storage diseases — problems beyond childhood
Authors: Lee P. J. , Leonard J. V.
Published: November 2004

The introduction of continuous nocturnal enteral glucose feeds and uncooked cornstarch has improved the prognosis for patients with the hepatic glycogen storage diseases. An increasing number of patients are surviving into adulthood in better health, but still at some medical cost. In this review we examine bone mineralization, renal function, hepatic tumours, and vascular endothelial function in GSD I and cardiac function in GSD III. All females over the age of 5 years with GSD I, III, VI and IX had morphologically polycystic ovaries. Thirteen adult GSD I patients have been studied, and been found to have poor bone mineralization and marked renal glomerular and tubular dysfunction. More than half of these patients also had focal hepatic lesions on sonography and yet vascular endothelial function was preserved in the face of hyperlipidaemia. In 12 GSD III patients, one had a focal hepatic lesion and 6 had pronounced left ventricular hypertrophy, although cardiorespiratory function was normal. These data emphasize the multisystem nature of these disorders and highlight the need for careful long‐term follow‐up.
Introduction to the age‐related diagnosis (ARD) index: an age at presentation related index for diagnostic use
Authors: Harkness R. A. , Harkness E. J.
Published: December 2004

Development and, therefore, age is important in paediatrics. Diagnostically useful data from this journal has been related to age and organized to form an age‐related diagnostic (ARD) index. The ARD index is designed for non‐expert clinical and laboratory workers to use in the early phases of diagnosis and as an addition to existing diagnostic schemes. Entry to the index is from the age at clinical presentation. Each entry is a sequence starting with clinical and laboratory presentations, clinical course, laboratory key investigations and finally diagnosis with volume and page numbers of the original article, the primary source. Within age groups, entries are grouped by diagnoses with the commonest diagnosis first; this has the effect of roughly but not precisely grouping similar clinical and laboratory findings.
Haplotype distribution at the phenylalanine hydroxylase locus in PKU families from the Moravian area of Czechoslovakia
Authors: Kozák L. , Dvořáková D. , Pijáčková A. , Kamarýt J.
Published: December 2004

The analysis of 21 families affected with classical phenylketonuria (PKU) from the Moravian area of Czechoslovakia has revealed 12 different RFLP haplotypes. Nine and eight haplotypes were associated with the normal and with the mutant alleles, respectively. Most normal alleles are associated with haplotype 1 (42.9%). Almost 80% of all mutant alleles are confined within only three haplotypes (1, 2 and 4). There was a strong association between haplotype 2 and the Czech mutant alleles (61.9% of the mutant alleles compared with 4.8% of the normal alleles). There was linkage disequilibrium between this haplotype and the R408W mutation in exon 12. Two mutant haplotypes 7 were found and in both cases they were tightly linked with G272ter mutation.

Our finding is in agreement with observations in other Eastern European countries. These data provide further support for the theories of the spread of the R408W mutation from east to west in European populations.
Heterozygotes for plasmalemmal carnitine transporter defect are at increased risk for valproic acid‐associated impairment of carnitine uptake in cultured human skin fibroblasts
Authors: Tein I. , DiMauro S. , Xie Z. ‐W. , De Vivo D. C.
Published: November 2004

One of the mechanisms by which chronic valproic acid (VPA) therapy induces serum and tissue depletion of carnitine in normal controls is through inhibition of plasmalemmal carnitine uptake (Tein et al 1993). To determine the effect of VPA on proven heterozygotes for the plasmalemmal carnitine transporter defect, we studied this system in cultured human skin fibroblasts with reducedVmaxfor the carnitine transporter usingL‐[3H]carnitine. There was en exponential dose‐dependent decrease in carnitine uptake with increasing VPA concentrations and the relative inhibitory effect was the same for all three carnitine concentrations for a given cell line. Importantly, the lower the maximal velocity of carnitine uptake of the heterozygote, the lower the number of carnitine transporters and the lower the carnitine uptake per given concentration of VPA. The degree of inhibition was also directly proportional to the time of VPA preincubation up to a specific maximal saturation time. The maximal effect of VPA exposure time was reached by 10 days in the control cell line and by 3 days in the two heterozygote lines, probably reflecting earlier saturation. We conclude that patients who are heterozygous for the plasmalemmal carnitine transporter defect are at increased risk for VPA‐associated serum and tissue depletion of carnitine through inhibition of plasmalemmal carnitine uptake.
Metachromatic leukodystrophy among southern Alaskan Eskimos: molecular and genetic studies
Authors: Pastor‐Soler N. M. , Schertz E. M. , Rafi M. A. , de Gala G. , Wenger D. A.
Published: November 2004

Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder resulting from the inability to metabolize sulphatide, an important component of myelin. Although there is significant clinical variability between patients, most have the late‐infantile form. It is one of the most common lysosomal disorders involving mental deterioration and is found throughout the world. The great majority of the cases have a deficiency of arylsulphatase A activity. Accurate diagnosis of MLD is complicated by the presence of so‐called pseudodeficiency alleles and the need to receive specimens for biochemical testing within 24–48h of collection. We report the identification of the mutation (a g‐to‐a transition in the first nucleotide of intron 4) in the arylsulphatase A gene causing late‐infantile MLD among the Eskimo population of southern Alaska. As all patients and family members from living and deceased patients had the same mutation, a mutation‐based test was developed to identify patients and carriers that can be done on dried blood spots sent via regular mail service. A possible genetic link between this population and the Navajo Indians of the southwestern United States is proposed.
Phenylketonuric patients decades after diet
Authors: Fisch R. O. , Chang P. ‐N. , Weisberg S. , Guldberg P. , Güttler F. , Tsai M. Y.
Published: November 2004

Nineteen early‐treated phenylketonuric patients, whose diet was discontinued between 4.5 and 13 years of age, and who have been off the diet for 12–28 years, were reassessed in 1992–93. There was little change in mean IQ between end of diet and follow‐up, less than one IQ point on the average, with no change for any individual exceeding 12 IQ points. Both prior and current IQ correlated slightly negatively with mean phenylalanine (Phe) concentration, and positively with parents' education. The phenylalanine level at follow‐up was significantly lower on average by about 900 µmol/L. Five of the subjects (26%) have evidence of mental disease. However, the data suggest that the discontinuation of the diet did not cause intellectual deterioration. Nonetheless, the patients' intellect cannot be the only consideration for maintenance of diet. The occurrence of psychopathology among phenylketonuric patients and the possible unknown effects of toxic elevation of phenylalanine during their lifetime suggest the need to maintain the diet. The use of DNA for diagnostic and prognostic purposes might assist in decisions about dietary quality and duration, and in anticipation of psychopathology.
Mitochondrial 2‐methylacetoacetyl‐CoA thiolase deficiency: An inborn error of isoleucine and ketone body metabolism
Authors: Søvik O.
Published: November 2004

A review is presented of 22 published cases of verified or probable mitochondrial 2‐methylacetoacetyl‐CoA thiolase deficiency, a disorder of isoleucine and ketone body metabolism. The clinical expression, characterized by ketoacidosis, vomiting and lethargy, is highly variable. Typical age of onset is between 6 and 24 months. The disorder, which has been observed in several ethnic groups, is apparently inherited as an autosomal, recessive trait. The prognosis is relatively good if acute episodes of ketoacidosis and dehydration are adequately treated. There is abnormal urinary excretion of 2‐methyl‐3‐hydroxybutyric acid, tiglylglycine, and in some instances also 2‐methyl‐acetoacetic acid. However, such a pattern of organic aciduria has also been found in cases with normal thiolase activity. Genetic complementation analyses have demonstrated considerable heterogeneity. The cDNA for human methyl‐acetoacetyl‐CoA thiolase has been cloned and sequenced. Studies in one patient showed a G‐to‐A substitution at position 1138 of the mRNA, causing 347Ala to Thr change in the mature enzyme. Studies in other patients have shown variable enzyme amount and/or stability.
A possible case of transient hereditary fructose intolerance
Authors: Catto‐Smith A. G. , Adams A.
Published: November 2004

A patient is described who presented with the signs and symptoms of hereditary fructose intolerance a few hours after her first fructose challenge. The diagnosis was confirmed by the demonstration of reduced activity of hepatic aldolase B towards fructose‐1‐phosphate. A second liver biopsy 10 months later had normal aldolase B activity towards fructose‐1‐phosphate and a fructose tolerance test was also normal. A possible explanation for these findings is proposed.
The effect of dietary fruits and vegetables on urinary galactitol excretion in galactose‐1‐phosphate uridyltransferase deficiency
Authors: Berry G. T. , Palmieri M. , Gross K. C. , Acosta P. B. , Henstenburg J. A. , Mazur A. , Reynolds R. , Segal S.
Published: November 2004

Even on a lactose‐restricted diet, urinary galactitol excretion and erythrocyte galactose‐1‐phosphate levels are persistently elevated in patients with galactose‐1‐phosphate uridyltransferase deficiency. In order to determine the contribution of galactose in dietary fruits and vegetables to this phenomenon, (1) the content of galactose in a lactose‐free diet was directly measured when a galactosaemic patient's diet was specifically enriched in those fruits and vegetables which contain relatively large amounts of free galactose and (2) galactitol excretion was determined during ingestion of this diet for 3 weeks and while on a synthetic diet for 1 week that provided <8 mg galactose/day. For comparison the effect of a 3‐week supplementation of 200 mg galactose/day was determined. The measured intake in total foodstuffs matched the theoretical content of galactose in the patient's diet based on amounts in fruits and vegetables alone, thus supporting the concept that fruits and vegetables are primarily responsible for galactose intake in a lactose‐free diet. All of the dietary manipulations, however, had relatively little effect on metabolite levels, suggesting that endogenous galactose production is primarily responsible for the elevated levels of galactose metabolites routinely detected in patients on lactose‐restricted diets.
The effect of lovastatin on very low‐density lipoprotein apolipoprotein B production by the liver in familial combined hyperlipidaemia
Authors: Cortner J. A. , Bennett M. J. , Le N. ‐A. , Coates P. M.
Published: November 2004

Overproduction of very low‐density lipoprotein apolipoprotein B by the liver is a metabolic marker for familial combined hyperlipidaemia, a common inherited disorder of lipoprotein metabolism. Four subjects with familial combined hyperlipidaemia had rates of apolipoprotein B production which were 2–7 times normal, using a protocol in which [15N]nglycine was used to label newly synthesized hepatic proteins. Following 4–6 months of therapy with lovastatin, very low‐density lipoprotein apolipoprotein B production in all four subjects had returned to the normal range. This demonstrates that lovastatin, an inhibitor of cholesterol biosynthesis, acts also to reduce the apparent production rate of apolipoprotein B by the liver.
Molecular basis of phenotype expression in homocystinuria
Authors: Kraus J. P.
Published: November 2004

Cystathionine β‐synthase (CBS) deficiency is the most common cause of homocystinuria in humans. The human gene maps to chromosome 21q22.3 and encodes the CBS subunit of 551 amino acid residues (63 kDa). CBS, a tetramer of these subunits, binds its two substrates, homocysteine and serine, and three additional ligands: pyridoxal 5′‐phosphate,S‐adenosylmethionine, and haem. Screening for mutations by expressing patient cDNA segments inE. coli permitted us to separate the parental CBS alleles, localize each mutation within one third of the cDNA, and functionally analyse the mutant protein. Using this method we identified the first 14 mutations in homocystinuria. The most common mutation in patients of predominantly ‘Celtic’ origin is the G919A transition which substitutes serine for glycine 307.
Organization of the human genome
Authors: Craig I. W.
Published: November 2004

The human genome contains about 3×109 base pairs which, based on average estimates for the size of coding regions, might suggest an upper limit of about 3 million genes. Evidence from a variety of approaches indicates a much lower figure — in the range 40 000–100 000 genes. Renaturation kinetic analysis reveals the presence of many repetitive elements in the human genome. Some of these are quite simple and highly reiterated, others are more complex and exist in fewer copies. The distribution and function of some of these families and the evolution of complex genomes from simpler ones are discussed.
Dysmorphic disorders — An overview
Authors: Donnai D.
Published: November 2004

Clinical delineation of dysmorphic syndromes is important for patient management, family counselling and basic research. Productive areas of research in dysmorphology and developmental biology have included the study of the mouse homologies of human disease. Mutations have been identified in both species in highly conserved ‘developmental’ genes, and also because of phenotypic similarity of syndromes. Mosaicism — somatic, germline and placental — involving chromosomal aneuploidy, single gene mutations and functional differences between cell lines is an important cause of malformations and syndromes. Many recurrent pattern malformation syndromes of previously unknown cause have now been found to be due to chromosomal microdeletions. Diagnosis has been greatly aided by the molecular cytogenetic technique of fluorescentin situ hybridization.
Primary hyperoxaluria type 1: Genotypic and phenotypic heterogeneity
Authors: Danpure C. J. , Jennings P. R. , Fryer P. , Purdue P. E. , Allsop J.
Published: November 2004

Primary hyperoxaluria type 1 (PH1) is an autosomal recessive disease caused by a deficiency of the liver‐specific peroxisomal enzyme alanine: glyoxylate aminotransferase (AGT). The disease is notable for its extensive heterogeneity at the clinical, biochemical, enzymic and molecular genetic levels. A study of 116 PH1 patients over the past 8 years has revealed four main enzymic phenotypes: (1) absence of both AGT catalytic activity and immunoreactive AGT protein (≈40% of patients); (2) absence of AGT catalytic activity but presence of immunoreactive protein (≈16% of patients); (3) presence of both AGT catalytic activity and immunoreactive protein (≈41% of patients), in most of which cases the AGT is mistargeted to the mitochondria instead of the peroxisomes; and (4) a variation of the mistargeting phenotype in which AGT is equally distributed between peroxisomes and mitochondria, but in which that in the peroxisomes is aggregated into matrical core‐like structures (≈3% of patients). Various point mutations, all occurring at conserved positions in the coding regions of the AGT gene, have been identified in these patients. The five mutations discussed in the present study, which have been found in individuals manifesting all of the four major enzymic phenotypes, account for the expressed alleles in about half of all Caucasian PH1 patients. The most common mutation found so far leads to a Gly170 → Arg amino acid substitution. This mutation, in combination with a normally occurring Pro11 → Leu polymorphism, appears to be responsible for the unprecedented peroxisome‐to‐mitochondrion mistargeting phenotype.
Variation in mitochondrial DNA levels in muscle from normal controls. Is depletion of mtDNA in patients with mitochondrial myopathy a distinct clinical syndrome?
Authors: Poulton J. , Sewry C. , Potter C. G. , Bougeron T. , Chretien D. , Wijburg F. A. , Morten K. J. , Brown G.
Published: November 2004

Recent studies have identified a group of patients with cytochrome oxidase (COX) deficiency presenting in infancy associated with a deficiency of mtDNA in muscle or other affected tissue (Moraes et al 1991). We used a novel approach to compare the level of mitochondrial (mtDNA) compared to nuclear DNA in skeletal muscle from a group of patients and controls, based on dot blots that were hybridized with a mtDNA probe labelled with35S[dCTP] and a reference nuclear DNA probe labelled with [32P]dCTP.

The ratio of mtDNA to nuclear DNA varied in samples from different muscles of the same individual. Secondly, fetal muscle had very low levels of mtDNA compared to nuclear DNA, and data from older controls (cross‐sectional rather than sequential) suggest that this increases rapidly over the first 3 months after birth and thereafter more slowly. Four patients with COX deficiency had levels of mtDNA that were below the age‐specific range defined by ‘normal’ quadriceps muscle. The clinical features of two of these patients were similar to earlier case reports of mtDNA depletion. In three patients the clinical course was relatively benign compared to cases that have previously been described.

Levels of mtDNA in skeletal muscle from some patients with other forms of muscle disease were also found to be low, suggesting that mtDNA depletion, possibly related to depletion of mitochondria, may be a relatively non‐specific response of muscle to various pathological processes. However, there does appear to be a distinctive group of young patients with reduced cytochrome oxidase activity in muscle, in whom marked mtDNA depletion reflects the primary defect.
Assay of plasmalogens and polyunsaturated fatty acids (PUFA) in erythrocytes and fibroblasts
Authors: Dacremont G. , Vincent G.
Published: November 2004

The direct transesterification method of Lepage and Roy is described as used in our laboratory for the analysis of plasmalogens and polyunsaturated fatty acids in erythrocytes and cultured fibroblasts by gas chromatography. An overview is given of the plasmalogen ratios and docosahexaenoic acid concentrations from controls and patients with different peroxisomal disorders investigated in our laboratory.
Measurement of peroxisomal fatty acid β‐oxidation in cultured human skin fibroblasts
Authors: Wanders R. J. A. , Denis S. , Ruiter J. P. N. , Schutgens R. B. H. , van Roermund C. W. T. , Jacobs B. S.
Published: November 2004

One of the main functions of mammalian peroxisomes is the β‐oxidation of a variety of fatty acids and fatty acid derivatives, including very long‐chain fatty acids. Oxidation of these fatty acids is deficient in a number of different peroxisomal disorders, including the disorders of peroxisome biogenesis (Zellweger syndrome, neonatal adrenoleukodystrophy and infantile Refsum disease), X‐linked adrenoleukodystrophy and a number of other disorders of peroxisomal β‐oxidation of known and unknown aetiology. Accurate measurement of peroxisomal fatty acid oxidation is of utmost importance for correct postnatal and prenatal diagnosis of these disorders. In this paper we describe a straightforward and accurate assay method to measure the β‐oxidation of palmitic acid (C16:0), hexacosanoic acid (C26:0) and pristanic acid in intact fibroblasts.
Activity measurements of acyl‐CoA oxidases in human liver
Authors: Van Veldhoven P. P.
Published: November 2004

Peroxisomal β‐oxidation is involved in the degradation of different fatty acids or fatty acid derivatives including eicosanoids (prostaglandins, leukotrienes, thromboxanes), dicarboxylic fatty acids, very long‐chain fatty acids, pristanic acid, bile acid intermediates (di‐ and trihydroxycoprostanoic acids), and xenobiotics. Separate β‐oxidation systems are probably active inside peroxisomes, each acting on a distinct set of substrates, as suggested by the discovery of multiple acyl‐CoA oxidases.

Using specific substrates or selective conditions, we can distinguish in rat liver the action of acyl‐CoA oxidases (type I and II), a pristanoyl‐CoA oxidase and a trihy‐droxycoprostanoyl‐CoA oxidase and, in human liver, of acyl‐CoA oxidase (type I and II) and a branched‐chain acyl‐CoA oxidase. When incubated with suitable CoA‐esters, these different oxidases can be measured in a similar fashion by following fluorimetrically the dimerization of homovanillic acid, catalysed by peroxidase in the presence of hydrogen peroxide. The optimal assay conditions and possible pitfalls in this type of coupled assay are discussed. This knowledge can be used to reveal the existence of peroxisomal disorders in which only one acyl‐CoA oxidase is deficient.
Liver and chorion cytochemistry
Authors: Roels F. , De Prest B. , De Pestel G.
Published: November 2004

Microscopic visualization of peroxisomes in chorionic villus cytotrophoblast and in biopsy and autopsy samples of liver and kidney, the presence of enlarged liver macrophages containing lipid droplets insoluble in acetone and n‐hexane as well as polarizing inclusions formed by stacks of trilamellar sheets are of diagnostic value in peroxisomal disorders. Methods are presented for evaluating these structures by light microscopy; trilamellar inclusions are only detected by electron microscopy. Macrophage features are preserved in archival paraffin blocks. In adrenal cortex, insoluble lipid, polarizing inclusions and trilamellar structures should be looked for. The stains are easily reproducible, and all reagents are commercially available.