bluuebunny/crossref_metadata_2025 huggingface dataset:
Ultrastructural and immunohistochemical features of lobular carcinoma of the breast
Authors: Nesland J. M. , Holm R. , Johannessen J. V.
Published: June 2005

Twenty cases of infiltrating lobular carcinoma (12 of classical type, five of trabecular type and three of alveolar type) and one case of lobular carcinoma in situ were studied by transmission electron microscopy. The in situ component in three of the infiltrating carcinomas was also studied. The ultrastructure of the tumour cells in the alveolar variant of infiltrating lobular carcinoma was the same as seen in the tumour cells of lobular carcinoma in situ. The tumour cells in infiltrating lobular carcinoma of the classical and trabecular types had more irregular nuclei and were more organelle‐ and filament‐rich. Immunohistochemical staining for the presence of α‐lactalbumin, casein, prekeratin and carcinoembryonic antigen (CEA) was done. The presence of α‐lactalbumin was proved in 19 per cent of the cases, casein 81 per cent, CEA in 65 per cent and prekeratin in 90 per cent. The light microscopic separation of some subgroups of infiltrating lobular carcinoma may be difficult, in particular the distinction between the classical and the trabecular variants. Unfortunately, our study shows that these distinctions cannot easily be made either by electron microscopy or by light microscopic immunohistochemistry with antibodies against prekeratin, CEA, casein and α‐lactalbumin.
Sex cord‐stromal tumours of the ovary
Authors: Fox H.
Published: June 2005

The aim of this review is to give a reasonably concise resumé of our knowledge of the sex cord‐stromal tumours of the ovary. Lipoid cell tumours of the ovary are often included within this broad category but this poorly defined and heterogenous group of neoplasms will not be considered here. This review is a selective one and no attempt is made to cover all aspects of sex cord‐stromal tumours or to provide a complete bibliography. The histological features of many of the neoplasms in this group, particularly those which have been recently defined, are discussed but a consideration of differential histological diagnosis is excluded. The ultrastructural characteristics of the various neoplasms are considered only in terms of their relevance to histogenesis or metabolic activity.
Copper toxicosis and tolerance in the rat. I‐changes in copper content of the liver and kidney
Authors: Haywood Susan
Published: June 2005

Rats can tolerate toxic copper diets; the extent of this adaptation and the changes that take place in the liver and kidney with respect to copper are studied. Rats were fed diets containing 3000–6000 mg/kg copper for 15 weeks and groups were killed at regular intervals. Their livers and kidneys were examined for pathological changes and for their copper content. Liver copper rose rapidly in the 3000 mg/kg trial to 4780 £ 636 μg/g copper between 4 and 5 weeks with marked liver damage. Stainable copper protein was present at 2 weeks in the hepatocytes but had disappeared by 5 weeks. Liver copper subsequently fell with recovery by 15 weeks. Kidney copper rose to plateau from 4 weeks. Stainable copper protein was present from 2 weeks in the cells of the proximal tubules and was apparently excreted. Tubular necrosis occurred at 4–5 weeks followed by regeneration. In the 6000 mg/kg trial the toxicosis was prolonged. Copper toxicosis in the rat can be a temporary phenomenon during the transformation of copper protein synthesis in the liver from a stainable to a non stainable form. Adaptation occurs after the removal of excess liver copper, facilitated probably by renal excretion, and the animals become tolerant.
The distribution of ceroid in human atherosclerosis
Authors: Mitchinson M. J. , Hothersall Deborah C. , Brooks P. N. , De Burbure Claire Y.
Published: June 2005

It has been suggested that macrophages in atherosclerotic plaques oxidize the lipid they contain, leading to necrosis in the plaque. Over 200 human aortic and coronary atherosclerotic plaques from 102 human necropsy subjects aged between 5 and 88 were, therefore, examined histologically for the presence of insoluble lipid (ceroid), thought to be a product of lipid oxidation. Ceroid was present in all the plaques but not in areas of diffuse intimal thickening. In early lesions the insoluble lipid was within membrane‐bound vesicles in macrophage‐like cells, many showing characteristic ring structures suggesting that membrane‐associated oxidative systems might be responsible for rendering the lipid insoluble. Staining was increased by an oxidizing agent and abolished by a reducing agent. It is suggested that this distribution supports the concept of lipid oxidation by macrophages within the plaque.
Immunohistochemical studies of c‐reactive protein and apolipoprotein b in inflammatory and arterial lesions
Authors: Rowe I. F. , Walker Louise N. , Bowyer D. E. , Soutar Anne K. , Smith L. C. , Pepys M. B.
Published: June 2005

Interactions in vivo between C‐reactive protein (CRP) and apolipoprotein B (apoB)‐containing lipoproteins were sought in inflammatory lesions and atherosclerosis. CRP was demonstrated immunohistochemically on the surface of some muscle fibres in locally induced inflammatory lesions in the rabbit, but apoB was not detected in the same distribution. CRP was not detected in catheter‐induced aortic endothelial injuries in the rabbit, in arterial lesions containing apoB from cholesterol‐fed rabbits, in apoB‐containing human fatty streaks or in advanced human atherosclerotic lesions.
Serotonin in fore‐gut carcinoids. A survey of 60 cases with regard to silver stains, formalin‐induced fluorescence and serotonin immunocytochemistry
Authors: Wilander E. , Lundqvist M. , El‐Salhy M.
Published: June 2005

A series of 60 fore‐gut carcinoid tumours was examined with regard to serotonin content after application of three different techniques, namely: the argentaffin reaction, formalin‐induced fluorescence according to Falck‐Hillarp and immunocytochemistry with monoclonal antibodies to serotonin. To evaluate the staining‐fluorescence of individual tumour cells, the methods were applied to identical tumour sections. Twelve tumours demonstrated serotonin‐immunoreactive cells, six of which were also argentaffin. Four tumours contained argentaffin cells but no serotonin immunoreactivity. With the use of all three techniques, three types of tumour cells occurred, namely: (1) serotonin‐immunoreactive, non‐argentaffin and non‐fluorescent cells, (2) serotonin‐immunoreactive, argentaffin and fluorescent cells, and (3) non‐serotonin immunoreactive, argentaffin and non‐fluorescent cells. The first (serotonin‐immunoreactive) cell type was most frequently found in the tumours. One gastric carcinoid in which the argentaffin cells exceeded the serotonin‐immunoreactive cells, a positive reaction was found with the modified Warthin‐Starry reaction for demonstrating melanin. Since none of the techniques used for visualization of serotonin in endocrine tumours is unquestionably specific and since they do not give identical results, it is indicated that for a more accurate identification of serotonin in fore‐gut carcinoid tumours, a positive reaction with at least two of the applied techniques is desirable.
Surface membrane phenotypic expression and treatment response of malignant lymphomas
Authors: Lauder I. , Bird C. C. , Child J. A. , Grigor I.
Published: June 2005

Cryopreserved lymphoid tissue has been used from 113 patients for an analysis of cell surface membrane phenotypes using a panel of monoclonal antibodies. Dual heavy chain expression was common in many B‐cell malignant lymphomas and in the B‐cell lymphomas as a whole there was a predominance of lambda light chains. The expression of lambda light chain was associated with a poorer response to treatment and there was a tendency for such tumours to fall into the high grade lymphoma category. Surface membrane heavy chain expression, as assessed by staining intensity, was greatest in the lymphomas composed mostly of centrocytes and IgD expression was reduced in high grade lymphomas. The ratio of helper to suppressor T‐cells was reduced in centroblastic and immunoblastic lymphomas. In Hodgkin's disease a predominance of T‐cells was found in most instances although the series included a large proportion of nodular sclerosis cases.
The binding of peroxidase‐labelled lectins to human breast epithelium. IV—the reactivity of breast carcinomas to peanut, soy bean and Dolichos biflorus agglutinins
Authors: Walker Rosemary A.
Published: June 2005

The binding of the lectins from peanut, soy bean and Dolichos biflorus (specific for galactose and/or N‐acetyl‐D‐galactosamine), to human breast carcinomas has been investigated. In contrast to the consistent pattern found in normal breast the reactivity of carcinomas to peanut lectin is varied, the most striking differences being prominent binding without desialation in some tumours and virtual lack of reactivity in others. There tends to be a greater degree of binding of soy bean agglutinin to carcinomas than normal although this is not absolute, and the reverse applies to Dolichos biflorus agglutinin. Comparison of the binding of the three lectins has revealed a complex heterogeneity of the carbohydrate chains within individual tumours and between different carcinomas. A significant relationship between peanut lectin binding, with the use of neuraminidase, and tumour differentiation has been found, and to a lesser extent with soy bean agglutinin when desialation is performed. However, the pattern of reactivity of the carcinomas with these three lectins does not appear to be related to local lymph node metastasis.
Cell proliferation within Bowman's capsule in mice
Authors: Morley A. R. , Wheeler Janice
Published: June 2005

Crescentic glomerulonephritis in man, irrespective of aetiology, indicates severe glomerular damage and carries a poor prognosis. The importance of the presence of macrophages in the development of glomerular crescents is well established, but the contribution of cellular proliferation in both macrophages and epithelial cells has received little attention. The purpose of this investigation was to develop a model of crescentic glomerulonephritis in mice, which will be suitable for cell kinetic studies. Preliminary studies are described, involving immunization with rabbit anti‐mouse glomerular basement membrane (GBM) antiserum after pre‐immunization with rabbit immunoglobulin G, both with and without treatment with a fibrinolytic inhibitor, Cyklokapron. Extensive glomerular damage, with crescent formation, was induced with four daily intravenous injections of rabbit anti‐mouse GBM antiserum, after pre‐immunization with rabbit IgG. The effect was shown to be dose dependant and was seen within 3 days of the last injection. Linear deposits of rabbit IgG were detected in all glomeruli by immunofluorescence and both linear and granular deposits of mouse IgG were also found. There was a significant increase in glomerular size and in the numbers of cells in Bowman's capsule (P < 0.01), and increases in mitotic indices to 0·8 per cent were seen by day 13.
Monoclonal antibodies specific for subsets of epidermal keratins: Biochemical and immunocytochemical characterization—applications in pathology and cell culture
Authors: Knight Jennifer , Gusterson Barry , Jones Robin Russell , Landells William , Wilson Patricia
Published: June 2005

Keratin composition has been widely used as a biochemical marker of differentiation in normal epithelia, cell culture systems and tumours of epithelial tissues. We have been developing a model system for the study of human squamous epithelial cell differentiation, and among a panel of monoclonal antibodies we have generated for analysing this system are two antibodies recognizing subsets of epidermal keratins. The two antibodies, designated LICR‐LON‐16a and LICR‐LON‐29b, were raised to the human squamous carcinoma cell line LICR‐LON‐HN‐5, and we describe here their biochemical and immunocytochemical characterization. Antibody 16a reacts with only epidermal basal cells in normal human skin and shows specificity for the 45 and 46 kdalton keratins. Antibody 29b stains all living layers of the epidermis, and reacts with a broad range of ketain polypeptides, (45–56 kdaltons) in immunoblotting analyses. We have investigated the alterations of cellular staining that occur in chronic hyperproliferative skin diseases and carcinomas and compared this with the staining of multilayered cultures of normal keratinocytes and the HN‐5 cell line. We show that in squamous cell carcinomas and in HN‐5 cell xenografts 16a and 29b stain only the well‐differentiated cell types. Furthermore we found that the basal cell specificity of 16a was lost in all of the hyper‐proliferative skin lesions examined including psoriasis and eczema. This transition to suprabasal staining pattern was also seen in the cultures of normal keratinocytes and HN‐5 cells. We conclude that aberrant keratin synthesis or abnormal post‐translational processing of keratins associated with an increased rate of cell turnover could account for the altered expression of the epitope recognized by antibody 16a.
G‐protein‐coupled receptor domain overexpression in Halobacterium salinarum: Long‐range transmembrane interactions in heptahelical membrane proteins
Authors: Jaakola Veli‐Pekka , Rehn Maria , Moeller Martina , Alexiev Ulrike , Goldman Adrian , Turner George J.
Published: June 2005

The aminergic α2b‐adrenergic receptor (α2b‐AR) third intracellular loop (α2b‐AR 3i) mediates receptor subcellular compartmentalization and signal transduction processes via ligand‐dependent interaction with Gi‐ and Go‐ proteins. To understand the structural origins of these processes we engineered several lengths of α2b‐AR 3i into the third intracellular loop of the proton pump bacteriorhodopsin (bR) and produced the fusion proteins in quantities suitable for physical studies. The fusion proteins were expressed in the Archaeon Halobacterium salinarum and purified. A highly expressed fusion protein was crystallized from bicelles and diffracted to low resolution on an in‐house diffractometer. The bR‐α2b‐AR 3i203–292 protein possessed a photocycle slightly perturbed from that of the wild‐type bR. The first half of the fusion protein photocycle, correlated with proton release, is accelerated by a factor of 3, whereas the second half, correlated with proton uptake, is slightly slower than wild‐type bR. In addition, there is a large decrease in the pKa, (from 9.6 to 8.3) of the terminal proton release group in the unphotolyzed state of bR‐α2b‐AR 3i as deduced from the pH‐dependence of the M‐formation. Perturbation of a cytoplasmic loop has thus resulted in the perturbation of proton release at the extracellular surface. The current work indicates that long‐range and highly coupled intramolecular interactions exist that are capable of “transducing” structural perturbations (e.g., signals) across the cellular membrane. This gene fusion approach may have general applicability for physical studies of G‐protein‐coupled receptor domains in the context of the bR structural scaffold. Proteins 2005. © 2005 Wiley‐Liss, Inc.
Flexibility of prolyl oligopeptidase: Molecular dynamics and molecular framework analysis of the potential substrate pathways
Authors: Fuxreiter Monika , Magyar Csaba , Juhász Tünde , Szeltner Zoltán , Polgár László , Simon István
Published: June 2005

The flexibility of prolyl oligopeptidase has been investigated using molecular dynamics (MD) and molecular framework approaches to delineate the route of the substrate to the active site. The selectivity of the enzyme is mediated by a seven‐bladed β‐propeller that in the crystal structure does not indicate the possible passage for the substrate to the catalytic center. Its open topology however, could allow the blades to move apart and let the substrate into the large central cavity. Flexibility analysis of prolyl oligopeptidase structure using the FIRST (Floppy Inclusion and Rigid Substructure Topology) approach and the atomic fluctuations derived from MD simulations demonstrated the rigidity of the propeller domain, which does not permit the substrate to approach the active site through this domain. Instead, a smaller tunnel at the inter‐domain region comprising the highly flexible N‐terminal segment of the peptidase domain and a facing hydrophilic loop from the propeller (residues 192–205) was identified by cross‐correlation analysis and essential dynamics as the only potential pathway for the substrate. The functional importance of the flexible loop has been also verified by kinetic analysis of the enzyme with a split loop. Catalytic effect of engineered disulfide bridges was rationalized by characterizing the concerted motions of the two domains. Proteins 2005. © 2005 Wiley‐Liss, Inc.
α1‐antitrypsin deficiency and liver disease: Clinical presentation, diagnosis and treatment
Authors: Hussain M. , Mieli‐Vergani G. , Mowat A. P.
Published: June 2005

The α1‐antitrypsin deficient subject (protease inhibitor (PI) phenotype ZZ) has an increased susceptibility to liver disease. The condition is most commonly identified in early infancy as a conjugated hyperbilirubinaemia with hepatitis (11%) or a bleeding state due to vitamin K malabsorption (2%). 50% of cases have cirrhosis and 25% die in the first decade of life. A further 2% present with cirrhosis in later childhood. Adult males are at risk of hepatoma development with or without cirrhosis. Diagnosis is by isoelectric focussing or allele‐specific oligonucleotide hybridization. The treatment is that of cholestasis and cirrhosis including transplantation. The pathobiology of the deficiency state, the mechanism of liver damage and the vulnerability of the newborn liver are discussed in this review. A plea is made for a trial of infusions of α1‐antitrypsin in early infancy, as is used safely but without proven efficacy in the emphysematous PIZZ subject. Prospects of therapy by gene modification are also reviewed.
α1‐antitrypsin deficiency and liver disease
Authors: Birrer P. , McElvaney N. G. , Chang‐Stroman L. M. , Crystal R. G.
Published: June 2005

α1‐Antitrypsin (α1AT) deficiency, one of the most common lethal hereditary disorders among Caucasians, is associated with emphysema in adults, while in children it is associated with liver disease. Produced in the liver and released into the plasma, α1AT serves as the body's major inhibitor of neutrophil elastase, a powerful proteolytic enzyme capable of degrading extracellular structural proteins. The pathogenesis of the liver disease associated with α1AT deficiency is not as well understood, but is clearly linked to specific mutations in coding exons of the α1AT gene, and the resulting accumulation of α1AT within hepatocytes. At present, therapy for the liver disease associated with α1AT deficiency is symptomatic, with liver transplantation as a last resort. New strategies are being developed to suppress the accumulation of α1AT by transferring the normal gene into the liver.
Paediatric liver transplantation: Review of current experience
Authors: Buckels J. A. C.
Published: June 2005

During the 1980s the results of liver replacement in children improved dramatically, with 12‐month survival rates rising from around 20% to over 85% at the most experienced centres. This improvement has been due to several factors, including better patient selection and timing of transplantation, advances in immunosuppressive therapy, and developments in liver preservation. Moreover, the learning curve effect has contributed with advances both in surgical technique and in the rapid diagnosis and treatment of complications, including the need to retransplant patients in whom the first graft has been irreversibly damaged. One major development is the refinement of the anatomically reduced grafts where a larger, usually adult, graft is cut down to fit a child. This has allowed a greater number of children to be grafted, including emergency cases such as fulminant hepatic failure in whom there is insufficient time to wait for a size‐matched donor.
Services for thalassaemia as a model for cost‐benefit analysis of genetics services
Authors: Modell B. , Kuliev A. M.
Published: June 2005

Economic appraisal of genetics services is usually limited to consideration of financial costs and benefits, but this can generate misunder‐standings about the aims of these services. We propose a general framework for economic analysis that includes non‐financial costs and benefits, and the concept of genetic fitness.
β‐Glucuronidase deficiency: Enzyme studies in an affected family and prenatal diagnosis
Authors: Maire I. , Mandon G. , Zabot M. T. , Mathieu M. , Guibaud P.
Published: June 2005

Aβ‐glucuronidase deficiency found in serum, leukocytes and fibroblasts and an increased [35S]sulphate incorporation in fibroblasts led us to diagnose two cases of type VII mucopolysaccharidosis in one family. In spite of the wide distribution of activities in serum from controls, decreasedβ‐glucuronidase activity allowed us to demonstrate the heterozygous status of the parents and two other children. Following these studies, an antenatal diagnosis was performed when the mother was pregnant again; amniotic fluid and cultured amniotic cells were used for enzyme activity determination. A heterozygous fetus was suspected and confirmed after birth. The reliability of various biological materials for enzymatic diagnosis and existence of genetic variants in the normal population are discussed.
Evolution of the neuroimaging changes in fucosidosis type II
Authors: Terespolsky D. , Clarke J. T. R. , Blaser S. I.
Published: June 2005

We report on clinical and neuroradiological findings in two patients with fucosidosis type II; a 7‐year‐old Jordanian boy and a 3 1/2‐year‐old Anglo‐Canadian girl. This rare, autosomal recessive disorder is caused by deficiency of lysosomalα‐fucosidase and is manifested clinically by progressive mental and motor deterioration, coarse facies, growth retardation, recurrent infections, dysostosis multiplex, angiokeratoma corporis diffusum, visceromegaly and seizures. Cranial CT and magnetic resonance imaging showed density and signal abnormalities in the thalamus, globus pallidus and internal capsules bilaterally, as well as progressive CT density alterations in supratentorial white matter including the internal medullary laminae of the thalami and the internal capsules.
Congenital nephrotic syndrome: A novel phenotype of type I carbohydrate‐deficient glycoprotein syndrome
Authors: van der Knaap M. S. , Wevers R. A. , Monnens L. , Jakobs C. , Jaeken J. , van Wijk J. A. E.
Published: June 2005

Type I carbohydrate‐deficient glycoprotein (CDG) syndrome is a genetic multisystem disorder generally without overt renal problems. We report a neonate with neurological abnormalities and congenital nephrotic syndrome of diffuse mesangial sclerosis type. Serum transferrin isoelectric focusing showed the typical abnormalities of type I CDG syndrome. Normal transferrin focusing findings in other patients with similar renal problems excluded the possibility of a secondary biochemical phenomenon. The diagnosis of type I CDG syndrome was confirmed by demonstration of a deficiency of phosphomannomutase. No evidence of pontocerebellar atrophy was found in imaging or at autopsy. We conclude that congenital nephrotic syndrome may occur in type I CDG syndrome, and that this diagnosis should be considered in patients with congenital nephrotic syndrome. Absence of pontocerebellar atrophy does not exclude the diagnosis of type I CDG syndrome.
Deficiency of pyrimidine 5′‐nucleotidase in human leukocytes
Authors: de Korte D. , van Doorn C. C. H. , Sijstermans J. M. J. , van Gennip A. H. , Roos D.
Published: June 2005

Both erythrocytes and leukocytes from a patient with erythrocyte pyrimidine 5′‐nucleotidase (P5N) deficiency were shown to contain increased amounts of pyrimidine nucleotides. These findings suggested that the leukocytes were also deficient for P5N. Measurement of the P5N activity in lysates from lymphocytes or granulocytes, in the presence of inhibitors for non‐specific 5′‐nucleotidase or alkaline phosphatase, indeed showed a deficiency for P5N in lymphocytes and granulocytes of the patient with erythrocyte P5N deficiency. However, the P5N deficiency in the leukocytes did not cause clinical disturbances in addition to the weak haemolytic anaemia.
Inheritance and prevalence of von Willebrand's disease severe form in a Brazilian population
Authors: Fischer R. R. , Lerner C. , Bandinelli E. , Fonseca A. S. K. , Roisenberg I.
Published: June 2005

Reviewed data suggest that the prevalence of severe von Wille‐brand's disease is influenced by ethnic and geographic factors. In the State of Rio Grande do Sul, Brazil, seven genealogies in which 11 individuals had a severe expression of von Willebrand's disease were localized. These affected subjects had similar laboratory results and all of them seemed to have resulted from double genetic defects, but the genealogic examination revealed that four of them probably resulted from combinations of autosomal recessive genes, while in the remaining the presence of dominant genes was likely and the involvement of genes for types I or II of von Willebrand's disease was possible. All of their examined relatives were asymptomatic but some of them presented unusual laboratory results, indicative of heterozygosis. The prevalence of severe cases in the surveyed population was higher than expected even when only the recessive forms were considered. It entered the expected values when it was presumed that these were all the cases currently living in the State. Genetic heterogeneity of the severe form was confirmed and it is suggested that the designations ‘severe von Willebrand's disease’ and ‘type III von Willebrand's disease’ should not be used as synonyms.
Crigler‐Najjar type II disease inheritance: A family study
Authors: Labrune P. , Myara A. , Hennion C. , Gout J. P. , Trivin F. , Odievre M.
Published: June 2005

The inheritance of Crigler‐Najjar type II disease is still contested. Autosomal dominant transmission with incomplete penetrance and autosomal recessive transmission have been proposed. We had the opportunity to study the hepatic activity of bilirubin uridinediphosphate glucuronyltransferase in parents whose first child had been affected by Crigler‐Najjar type II disease. The demonstration of reduced activity of glucuronidation in the liver of both parents suggests autosomal recessive inheritance. The second infant of this couple was affected by the same disease and was treated with success by phenobarbital.
Holocarboxylase synthetase deficiency: 9‐year follow‐up of a patient on chronic biotin therapy and a review of the literature
Authors: Michalski A. J. , Berry G. T. , Segal S.
Published: June 2005

We report on the long‐term medical and neurodevelopmental follow‐up of a patient with the rare and potentially lethal disease, holocarboxylase synthetase deficiency. He was originally treated prenatally with biotin megatherapy and for 9 years with 6 mg/day since his only episode of fulminant acidosis at 3 months of age. While growth and general health have been normal, the patient has exhibited signs of minimal brain dysfunction. However, evaluation of unaffected siblings suggests that this may be unrelated to his metabolic disease. A review of the literature and recommendations for optimal treatment are provided.
Association of hyperprolinaemia type I and heparin cofactor II deficiency with CATCH 22 syndrome: evidence for a contiguous gene syndrome locating the proline oxidase gene
Authors: Jaeken J. , Goemans N. , Fryns J. ‐P. , François I. , de Zegher F.
Published: June 2005

Increased proline levels were found in plasma of a girl with slight psychomotor retardation, epilepsy, obesity, scoliosis, hypocalcaemia, variable lymphocytopenia and facial dysmorphy suggestive of CATCH 22 syndrome. Fluorescencein situ hybridization indicated the presence of a submicroscopic 22q11 deletion, confirming this diagnosis. Further investigation showed evidence that the patient was heterozygous for heparin cofactor II deficiency and for hyperprolinaemia type I, a proline catabolic disorder due to proline oxidase deficiency. This association extends the CATCH 22 syndrome and suggests that expression of the proline oxidase gene depends on the chromosome 22q11 region.
Mucopolysaccharidosis IVA: a comparative study of polymorphic DNA haplotypes in the Caucasian and Japanese populations
Authors: Rezvi G. M. M. , Tomatsu S. , Fukuda S. , Yamagishi A. , Cooper A. , Wraith J. E. , Iwata H. , Kato Z. , Yamada N. , Sukegawa K. , Shimozawa N. , Suzuki Y. , Kondo N. , Orii T.
Published: June 2005

Mucopolysaccharidosis IVA (MPS IVA) is an autosomal recessive disorder caused by deficiency of the lysosomal enzymeN‐acetylgalactosamine‐6‐sulphate sulphatase (GALNS). The genetic heterogeneity at the GALNS locus was studied in 62 mutant alleles and 376 normal alleles in the Caucasian population and also in 40 mutant and 100 normal alleles in the Japanese population. For this study, six different restriction fragment length polymorphisms (RFLPs) at the GALNS locus were analysed to search for the frequency of each RFLP produced byStyI,SphI,RsaI,HaeIII,StuI andHapII restriction endonucleases. We detected a total of 27 haplotypes in the Caucasian and Japanese population. Of these 27 haplotypes, 18 haplotypes were present in the Caucasian population and the most common of these was haplotype 1 (ABHcde) in both mutant and normal alleles. In contrast, in the Japanese population we found 20 of the 27 haplotypes and the most common in mutant and normal alleles was haplotype 2 (abhcDE). Within these two populations a parent in the MPS IVA family has an average probability of greater than 77% (in the Caucasian population 77.27% and in the Japanese population 78.26%) of being heterozygous, and hence informative for linkage, at one or more GALNS RFLP sites. Our results delineate the molecular heterogeneity of MPS IVA haplotypes, as well as their significant interpopulation variation, and make prenatal diagnosis and carrier detection possible in the majority of families with one affected child.
Seizures in a boy with succinic semialdehyde dehydrogenase deficiency treated with vigabatrin (γ‐vinyl‐GABA)
Authors: Matern D. , Lehnert W. , Gibson K. M. , Korinthenberg R.
Published: June 2005

Succinic semialdehyde dehydrogenase (SSADH) deficiency is an autosomal recessive disease involving the catabolism of the neurotransmitter γ‐amino‐butyric acid (GABA). The main symptoms include retardation of psychomotor and language development, muscle hypotonia and non‐progressive ataxia. Therapy consisting of approximately 75 mg/kg per day of vigabatrin, an irreversible inhibitor of GABA‐transaminase, is reported to lead to some improvement of the clinical condition in affected patients. We report on a 12‐year‐old boy with SSADH deficiency who, when treated with 75 mg/kg per day of vigabatrin, showed marked amelioration of symptoms but also EEG changes and two generalized seizures. On discontinuing vigabatrin therapy, the seizures resolved and the EEG improved, but the patient's clinical condition deteriorated to its pre‐treatment state. A stable EEG without the recurrence of seizures as well as renewed improvement of cognitive and behavioural functions was achieved with a reduced vigabatrin dose of 25 mg/kg per day. We conclude that vigabatrin in SSADH deficiency should be administered in a gradually increasing dosage combined with frequent evaluation of the clinical condition and the EEG.
Homocysteine response to methionine challenge in four obligate heterozygotes for homocystinuria and relationship with cystathionineβ‐synthase mutations
Authors: Sperandeo M. P. , Candito M. , Sebastio G. , Rolland M. O. , Turc‐Carel C. , Giudicelli H. , Dellamonica P. , Andria G.
Published: June 2005

Fasting and post‐methionine load plasma total homocysteine concentrations were investigated in the parents of two homocystinuric patients. Three genetic mutations in the cystathionineβ‐synthase gene were found. In the patient of family 1, a frequent Caucasian mutation, T833C, was found on one allele, while the mutation on the other allele has not yet been defined. In the patient of family 2, a mutation C569T, recently described by Sperandeo and colleagues, was found on one allele, while a novel mutation, G346A, was characterized on the other allele. The frequent gene mutation T833C was detected in a heterozygous mother who, surprisingly, exhibited strictly normal fasting and post‐methionine load homocysteinaemia. In contrast, in the other family, we found a novel mutation (G346A) in the mother located near Lys 119, the putative binding site of phosphopyridoxal phosphate. This mother exhibited increased fasting and post‐methionine load homocysteinaemia. These observations could explain the conflicting results reported for vascular pathologies in parents of homocystinuric patients and direct the search for genetic mutations in these vascular pathologies.
Mucopolysaccharidosis type IVA (morquio syndrome): A clinical review
Authors: Northover H. , Cowie R. A. , Wraith J. E.
Published: June 2005

Patients with MPS IV have a clinical disorder quite different from other MPS conditions. The major treatment issue revolves around the prevention of cervical myelopathy, although the other aspects of this multisystem disease should not be forgotten. Management is inevitably multidisciplinary and the paediatrician should play a lead role in the coordination of services for affected patients.

The timing of occipito‐cervical fusion and the selection of patients for this procedure remain unclear. A prophylactic approach to surgery is suggested in this review, although it is by no means clear that all patients will invariably develop cervical myelopathy if left untreated.

Parents of affected children need considerable support in dealing with the affected child and the children themselves need help to try to come to terms with their severe physical disabilities. The Society for Mucopolysaccharide Diseases (55 Hill Avenue, Amersham, Bucks HP6 5BX, UK) has offered many parents and affected individuals considerable help as well as raising significant sums for research.

Finally, it is important to remember the genetic nature of the disease and the possibility of prenatal diagnosis in subsequent pregnancies.
The salience of Garrod's ‘molecular groupings’ and ‘Inborn Factors in Disease’
Authors: Scriver C. R.
Published: June 2005

Garrod's important second book,Inborn Factors in Disease (1931), was about inherited predisposition to disease. Chemical and metabolic individuality, which are the modalities of predisposition, originated in ‘molecular groupings’ (proteins) in Garrod's view of life. Such ‘groupings’ as interlocus molecular hybrids, allelic complementation and expressions of modifier genes, can assume variant expression in heterozygotes. Here, it is shown that genetic variation in such ‘molecular groupings’ has clinical relevance, for example (1) in reproductive counselling for thalassaemia; (2) in heterozygosity where the affected enzymes are normally homopolymeric; (3) in clinical severity of ‘monogenic’ disease (e.g. familial hypercholesterolaemia and muscular dystrophy) when variation is not explained by allelic heterogeneity. The associated chemical individuality in each case can be used to identify risk and thus as a mode of predictive medicine.
Acute metabolic encephalopathy: A review of causes, mechanisms and treatment
Authors: Surtees R. , Leonard J. V.
Published: June 2005

Acute encephalopathy is a relatively common problem: one of the causes is metabolic disorders. A detailed history, examination and investigations performed during the acute illness (blood sugar, blood gases, plasma ammonia, blood lactate, plasma ketones, plasma amino acids, liver function tests, and urinary organic acids) should identify those patients in whom a metabolic disorder is likely. More detailed studies may be needed to establish a precise diagnosis.

The mechanism of the acute brain dysfunction is multifactorial. Factors that contribute include changes in blood flow and, initially, a disturbance in neurotransmitter function followed by failure of energy metabolism and cellular depolarization. Treatment of these conditions is largely supportive, with especial attention to the management of cerebral perfusion pressure.
A clinical biochemist's view of the investigation of suspected inherited metabolic disease
Authors: Blom W. , Huijmans J. G. M. , van den Berg G. B.
Published: June 2005

The necessity for a multi‐disciplinary approach to the study of genetic disease is discussed. The progress of laboratory investigation programmes made it not feasible and inefficient to run a full metabolic investigation programme in every new patient suspected of inherited metabolic disease. An application form for metabolic investigation is described, which can be used to collect clinical information relevant to metabolic disease. On the basis of the patient's clinical information, selection criteria are given to decide which laboratory investigation programme has to be performed in the individual patient. A full metabolic laboratory investigation programme is described and illustrated with some examples of abnormal metabolite patterns. Diagnostic results over a 2‐year period are presented.
Genetic aspects of prenatal diagnosis
Authors: Connor J. M.
Published: June 2005

With improved control of environmental agents, genetic conditions are now a major cause of residual handicap and mortality in all age groups. Primary prevention of this diverse group of over 5000 distinct disorders is not yet possible and effective therapy is, as yet, available for very few. Hence, the present emphasis on prevention is directed towards the identification and testing of pregnancies at risk in order to allow the option of early selective termination of pregnancy. Although over 400 distinct conditions have already been successfully prenatally diagnosed, the identification of at‐risk pregnancies on the basis of a positive family history alone can identify only a minority of affected pregnancies. In contrast, genetic screening during (or before) pregnancy offers a real prospect for detection of a majority of affected pregnancies and hence for reducing the birth frequency of serious genetic conditions. This is exemplified by the impact of the maternal serum α‐fetoprotein screening programme for neural tube defects and is under active development for autosomal aneuploidies and certain other major congenital malformations. These screening programmes will result in a reduced frequency of mental and physical handicap in the community but their implementation will require a comprehensive team approach with supraregional (or national) funding and co‐ordination.
Obstetric aspects of prenatal diagnostic methods
Authors: Whittle M. J. , Gilmore D. H. , McNay M. B.
Published: June 2005

The vast improvements in ultrasound technology have proved to be an important starting point for invasive investigative procedures in obstetrics. These methods not only allow a more precise diagnosis to be made but also offer considerable scope forin utero therapy.
Prenatal and perinatal diagnosis of peroxisomal disorders
Authors: Schutgens R. B. H. , Schrakamp G. , Wanders R. J. A. , Heymans H. S. A. , Tager J. M. , van den Bosch H.
Published: June 2005

Peroxisomes play an essential role in human cellular metabolism. Peroxisomal disorders, a group of genetic diseases caused by peroxisomal dysfunction, can be classified into three groups: (1) disorders of peroxisome biogenesis with a generalized loss of peroxisomal functions (Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum disease, hyperpipecolic acidaemia); (2) disorders with a loss of multiple peroxisomal functions (rhizomelic chondrodysplasia punctata and Zellweger‐like syndrome; (3) disorders with loss of a single peroxisomal function (X‐linked adrenoleukodystrophy, peroxisomal thiolase deficiency, bifunctional protein deficiency, acyl‐CoA oxidase deficiency, classic Refsum disease, hyperoxaluria type I and acatalasaemia). Prenatal diagnosis is indicated in all these genetic disorders with the exception of classic Refsum disease, most types of hyperoxaluria type I and acatalasaemia.

A variety of techniques is available now for the prenatal diagnosis of peroxisomal disorders in the first or second trimester of gestation. Prenatal diagnosis was performed by us in 70 pregnancies at risk for a disorder of peroxisome biogenesis, three for rhizomelic chondrodysplasia punctata, four for X‐linked adrenoleukodystrophy and two for a defect in peroxisomal β‐oxidation. Fourteen affected fetuses were identified; no false negative cases were obtained.
Prenatal diagnosis of disorders of galactose metabolism
Authors: Holton J. B. , Allen J. T. , Gillett M. G.
Published: June 2005

Of three clinically significant galactose disorders, there is only a real need and experience of prenatal diagnosis in classical galactosaemia. Prenatal diagnosis for this disorder may be carried out by galactose‐1‐phosphate uridyl transferase assay in cultured amniotic fluid cells or in chorionic villus biopsies and by galactitol estimation in amniotic fluid supernatant. Although the long‐term outcome of patients treated on a galactose‐restricted diet is recognized to be unsatisfactory, prenatal diagnosis is only rarely performed with a view to terminating the affected pregnancy.
Disorders of mitochondrial β‐oxidation: Prenatal and early postnatal diagnosis and their relevance to Reye's syndrome and sudden infant death
Authors: Pollitt R. J.
Published: June 2005

There are still many problems with the diagnosis and classification of inherited disorders of mitochondrial β‐oxidation. At present only the acyl‐CoA dehydrogenase step of the β‐oxidation spiral has been explored in any detail and a large number of patients have disorders that cannot be properly characterized. β‐Oxidation defects may present in a wide variety of ways, the most dramatic being acute encephalopathy with hepatic involvement (atypical Reye's syndrome) or ‘sudden’ death. Investigations may include urinary and plasma organic acids, metabolic stress tests and assays of overall metabolic pathways or of specific enzymes in cultured fibroblasts, lymphocytes, or other material. Early postnatal diagnosis presents particular difficulties but in medium‐chain acyl‐CoA dehydrogenase deficiency the diagnosis may be apparent from careful examination of urine. There is as yet little general experience in prenatal diagnosis of this group of disorders except for glutaric aciduria type II. Single prenatal diagnoses of medium‐chain acyl‐CoA dehydrogenase deficiency and of an incompletely characterized defect of medium‐chain fatty acid oxidation have been performed.
The control of 5‐hydroxytryptamine and dopamine synthesis in the brain: A theoretical approach
Authors: Hommes F. A. , Lee J. S.
Published: June 2005

The transport of the eight amino acids (phenylalanine, tyrosine, tryptophan, valine, leucine, isoleucine, histidine and methionine) using the large neutral amino acid transporter of the blood‐brain barrier (BBB) has been calculated using published kinetic data. The fate of the amino acids has been followed from blood to interstitial space, to cell and through metabolism which included, for tyrosine and tryptophan, the hydroxylases. The system was analysed in terms of flux control coefficients. Since the summation theorem did not hold, the system clearly behaved as a non‐homogeneous system. At physiological levels of these eight amino acids, the largest contribution to the control of the flux of tyrosine is given by the hydroxylase step, followed by the diffusional component of the transport across the BBB. For tryptophan it is the hydroxylase step, followed by the carrier‐mediated transport across the BBB. For the other amino acids it is the metabolism, followed by the diffusional component of the BBB transport.

These parameters for tyrosine and tryptophan were determined at increased levels of blood phenylalanine, tyrosine or histidine. The flux through tryptophan hydroxylase can be affected by high blood levels of tyrosine and histidine to values also observed in hyperphenylalaninaemia. Since hypertyrosinaemia (type II) and hyperhistidinaemia are not associated with mental retardation, it is concluded that interference with transport across the BBB of tyrosine and tryptophan, as well as the flux through tryptophan hydroxylase leading to the synthesis of 5‐hydroxytryptamine, do not contribute to the cause of permanent brain dysfunction in hyperphenylalaninaemia.

It can be calculated that addition of tyrosine to the diet to raise the blood tyrosine level in phenylketonuria patients may have a beneficial effect for the synthesis of neurotransmitters derived from tyrosine.
Results of newborn screening for galactose metabolic disorders
Authors: Inoue B. , Hata M. , Ichiba Y. , Wada H. , Misumi H. , Mori T.
Published: June 2005

A screening strategy has been used which uses the Paigen and Beutler methods for the determination of galactose and galactose‐1‐phosphate. A blood spot test for epimerase has also been developed. In the last 10 years, 265019 samples from newborns have been tested by these methods. Among the 154 screening positives, we have detected seven cases of epimerase‐deficient galactosaemia (Type III), seven cases of Duarte/galactosaemia heterozygotes, 48 cases of other various types of heterozygotes, four cases of persistent hypergalactosaemia, three cases of hepatitis and one case of congenital atresia of the bile duct. These results indicate that our screening system has effectively detected the infants with galactose metabolic disorders.
Immunoquantification of the low abundance lysosomal enzymeN‐acetylgalactosamine 4‐sulphatase
Authors: Brooks D. A. , McCourt P. A. G. , Gibson G. J. , Hopwood J. J.
Published: June 2005

The low abundance lysosomal enzymeN‐acetylgalactosamine 4‐sulphatase (4‐sulphatase) has been quantified using a microimmunopurification step and a monoclonal‐based ELISA detection system. The assay is similar in principle to a two‐site ELISA but uses a single monoclonal antibody against one epitope to bind 4‐sulphatase in two separate assay steps. The sensitivity of this assay is sufficient to allow the quantification of 4‐sulphatase in human cultured skin fibroblasts derived from normal controls and patients deficient in 4‐sulphatase activity (mucopolysaccharidosis type VI or Maroteaux‐Lamy syndrome). The results obtained suggest a range of mucopolysaccharidosis type VI or 4‐sulphatase deficient mutants, from those expressing little or no quantifiable 4‐sulphatase protein to those examples with quantifiable levels of 4‐sulphatase protein which is enzymically inactive. Phenotypic variability in patients with a 4‐sulphatase deficiency may therefore be partially attributed to a range of protein expressions. The method should allow the determination of 4‐sulphatase specific activity in mucopolysaccharidosis type VI patients.
International database of tetrahydrobiopterin deficiencies
Authors: Blau N. , Barnes I. , Dhondt J. L.
Published: June 2005

Approximately 2% of newborns with hyperphenylalaninaemia are deficient in tetrahydrobiopterin. Selective screening must be performed in all instances where hyperphenylalaninaemia is detected by neonatal screening. In the last 20 years, 308 patients with tetrahydrobiopterin deficiencies have been recognized as a result of screening carried out, worldwide, in Departments of Paediatrics. Of these 308 patients, 181 suffered from 6‐pyruvoyltetrahydropterin synthase deficiency, 92 from dihydropteridine reductase deficiency, 13 from pterin‐4a‐carbinolamine dehydratase deficiency, 12 from GTP cyclohydrolase I deficiency, and 10 are still unclassified. In the BIODEF database we have tabulated the most common clinical and laboratory data related to hyperphenylalaninaemia and tetrahydrobiopterin deficiencies. Additionally, there are data regarding treatment, outcome, and DNA analysis. Preliminary evaluation reveals that the degree of hyperphenylalaninaemia can vary from normal to 2500 µmol/L. Analyses of pterins in urine and measurement of dihydropteridine reductase activity from Guthrie cards are absolutely essential tests for accurate diagnosis. There is a regional (demographic) variation in the frequency of tetrahydrobiopterin deficiencies indicating the highest incidence in Saudi Arabia, probably a consequence of the high consanguinity rate.