bluuebunny/crossref_metadata_2025 huggingface dataset:
Dynamic contrast‐enhanced MRI for assessing the disease activity of multiple myeloma: A comparative study with histology and clinical markers
Authors: Nosàs‐Garcia Sílvia , Moehler Thomas , Wasser Klaus , Kiessling Fabian , Bartl Reiner , Zuna Ivan , Hillengass Jens , Goldschmidt Hartmut , Kauczor Hans‐Ulrich , Delorme Stefan
Published: June 2005

Purpose

To examine whether parameters of dynamic, contrast‐enhanced MRI (DCE MRI) reflect the degree of infiltration and vessel density in corresponding bone marrow biopsy specimens.

Materials and Methods

The pelvis of 24 patients with multiple myeloma (MM) was examined using contrast‐enhanced DCE MRI. Biopsy was obtained from the spina iliaca posterior superior. Using a two‐compartment model (assuming one intravascular and one interstitial compartment), the parameters amplitude (A, the maximum, relative signal increase over baseline) and the exchange rate constant (kep, describing the redistribution of contrast agent from the interstitial into the intravascular compartment) in the biopsied region were calculated and compared with the histological and clinical data.

Results

DCE MRI parameters were significantly higher in lesions with marked infiltration than with mild or no infiltration (P < 0.05). The amplitude normalized to that in the iliac artery (An), but not kep, was higher in lesions with high vessel‐density at histology (P = 0.01). Higher kep levels were found in presence of increased serum immunoglobulins.

Conclusion

Increased contrast uptake in the bone marrow of MM patients indicates at least moderate tumor involvement. Furthermore, the contrast enhancement correlates with vessel‐density and serum markers of disease activity. J. Magn. Reson. Imaging 2005;22:154–162. © 2005 Wiley‐Liss, Inc.
Excess plasma proteins as a cause of chronic inflammation and lymphoedema: Biochemical estimations
Authors: Gaffney R. M. , Casley‐Smith J. R.
Published: June 2005

The subcutaneous tissue of rats was injected with plasma, PVP or saline. Precautions were taken to avoid Immunological reactions and the release of mediators. The repeated injection of plasma proteins caused changes typical of chronic inflammation in the skin and to a much greater extent in the fascia. Coumarin significantly reduced the extent of this inflammatory reaction in the skin but not in the fascia. It increased the numbers of cells in the skin and may have stimulated them to lyse the excess protein. After 32 days, PVP injection caused some of the changes seen in chronic inflammation, but to a much lesser extent than those found with plasma. The only effect of coumarin was to reduce the water content of the skin (and hence the skin weight).

It is concluded that the accumulation of excess proteins in the tissues can indeed cause chronic inflammation. In particular, chronic lymphoedema can be regarded as a form of chronic inflammation. It is also concluded that in the presence of excess protein, the benzopyrone, coumarin, is capable of stimulating cells, other than macrophages, in the skin.
Transmission electron microscopy of human oesophageal carcinomas
Authors: Robinson K. M. , Gregory M. A.
Published: June 2005

Twelve human oesophageal carcinoma samples were examined by both light and electron microscopy in order to characterise the ultrastructural features of this neoplasm. Those samples from well differentiated tumours contained abundant tonofilaments, mainly in a paranuclear position, while cells from poorly differentiated tumours displayed few such filaments but had more prominent cytoplasmic organelles. Desmosomes were well developed in all samples, some of which displayed numerous clearly defined intracellular desmosomes not necessarily associated with cell membrane material. Myelinoid figures were abundant and the bizarre forms which were present in some samples may be related either to abnormal metabolism or to viral activity. Glycogen, lipid droplets and intermediate filaments, possibly related to basement membrane material were observed in several samples, No single feature diagnostic for carcinoma of the oesophagus emerged from this study, but several structural similarities between oesophageal and other carcinomas were noted.
The glomerular basal lamina in hereditary nephritis
Authors: Yoshikawa N. , Cameron A. H. , White R. H. R.
Published: June 2005

Characteristic ultrastructural alterations of the glomerular basal lamina have been reported in hereditary nephritis. The basal lamina is irregularly thickened and the lamina densa shows replication with a “basket weave” pattern, enclosing electron‐lucent lacunae which frequently contain small dense particles. However there is controversy regarding the specificity of this lesion in hereditary nephritis. To determine the specificity, 366 renal biopsies from 310 children were studied retrospectively. Twenty‐four out of 27 patients with hereditary nephritis showed the characteristic changes of the basal lamina and they were widespread in 17.

Two patients with recurrent haematuria but no family history of deafness or haematuria showed similar extensive changes and are regarded as new mutant cases of hereditary nephritis.

Similar changes were seen in 17 of the 281 patients with other conditions but were always localized to a few capillary loops. We conclude that a widespread “basket weave” pattern appears to be confined to hereditary nephritis and is seen in the great majority of such cases.
Furosemide induced hepatotoxicity
Authors: Walker R. M. , McElligott T. F.
Published: June 2005

Furosemide‐induced (400mg/kg ip) hepatotoxicity progressing to centrilobular necrosis was studied by light and electron microscopy in Swiss white mice. Centrilobular glycogen depletion and cytoplasmic foaminess, usually accompanied by extensive vacuolation, were detectable by light microscopy 1 1/2 hr after furosemide. Centrilobular congestion and hydropic single cell necrosis developed after 3 hr. Electron microscopy revealed disaggregation of polyribosomes, vesiculation of endoplasmic reticulum, an endocytic origin for the vacuolation, and a definite sequence in the development of congestion. Vacuolation coincided with loss of microvilli and resulted in detachment of sinusoidal lining cells from hepatocytes. Small vacuoles apparently formed at the sinusoidal margin of hepatocytes by fusion of microvilli. Congestion and occlusion of sinusoidal lumens developed as a consequence of erythrocytes entering the enlarged Disse space, possibly through pores in the lining cells, and thence entering endocytic vacuoles. The vacuolation, loss of microvilli and terminal hydropic degeneration suggest an important role for the plasma membrane in the development of furosemide‐induced hepatotoxicity. Comparisons with known membrane toxins support this concept.
Direct evidence that inflammatory multinucleate giant cells form by fusion
Authors: Murch A. R. , Grounds Miranda D. , Marshall C. A. , Papadimitriou J. M.
Published: June 2005

Multinucleate giant cells (MGC) are believed to be formed by fusion of macrophages. In a chimaeric mouse composed of two histoincompatible strains each homozygous for one of the two isoenzymic forms of glucose‐6‐phosphate isomerase it was found that hybrid enzyme was produced in MGC‐rich leucocytic exudates. This hybrid can only occur if nuclei of the two different strains reside within a common syncytial cytoplasm, demonstrating unequivocally that macrophage fusion occurred between cells of the two strains. Since the two strains were histoincompatible it appears that no strain specific recognition is necessary for fusion to occur.
Rat pancreas allotransplantation: A short term comparison of rejection patterns with different methods of exocrine drainage
Authors: Lindsey N. J. , Nolan M. S. , Slater D. N. , Savas C. P. , Boyle P. F. , Herold A. , Beck S. , Fox M.
Published: June 2005

Pancreas transplantation has been performed in the rat to study the effect on rejection of duct ligation, duct obliteration and duct open intraperitoneal drainage. On day six, an acceleration of graft damage was seen in the duct ligated and obliterated allografts compared to the duct open group. However, by day 11, rejection was complete in all three groups. In comparison with pancreatic isografts, the histological diagnosis of rejection can be difficult to define, particularly in the duct ligated and obliterated groups. The most useful criterion was the localisation of the cellular infiltrate to the walls of small veins and within the islets of Langerhans.
A quantitative histomorphologic analysis of lymph node granulomas in sarcoidosis in relation to radiological stage I and II
Authors: van Maarsseveen A. C. M. Th. , Veldhuizen R. W. , Stam J. , Alons C. L. , Mullink H.
Published: June 2005

In a detailed study of granulomas in lymph nodes of 19 patients with Sarcoidosis we assessed and correlated a number of features, with size, development and radiographic staging of the disease. Except with the epithelioid cells the number of constituent cells (lymphocytes, giant cells and polymorphs), size, confluency and degree of necrosis of the granulomas were found to be very variable within a single gland. This indicates that the granulomas are in different stages of activity at a given time. All the features studied showed little or no correlation with the radiographic staging, which suggests that the latter is not a good index of the extent of the disease.
The regression of oestradiol‐induced pituitary tumours in the rat
Authors: Treip C. S.
Published: June 2005

Wistar rats of both sexes were implanted subcutaneously with oestradiol pellets (25 mg). The pellets were removed in two groups of animals after 120 and 220 days of implantation; in other animals the pellets were left for 230 and 370 days. The animals in whom pellets were left implanted all had large pituitary tumours. The animals from whom the pellets had been removed had pituitaries larger than normal, but much smaller than those in which the implants had been left. The results suggest that oestradiol‐induced pituitary tumours are totally hormone‐dependent and are not autonomous neoplasms. Oestradiol‐induced testicular atrophy was also reversible.
The binding of peroxidase‐labelled lectins to human breast epithelium. III—altered fucose‐binding patterns of breast carcinomas and their significance
Authors: Walker Rosemary A.
Published: June 2005

Tissue sections from 80 human breast carcinomas have been examined for evidence of binding of the two fucose‐specific lectins, Lotus tetragonolobus and Ulex europeus I, with enzymes utilized as an indicator system. Both singlestaining and double‐staining methods (Lotus tetragonolobus‐peroxidase and Ulex europeus I‐alkaline phosphatase) have been employed. In contrast to normal breast in which there is consistent reactivity of all epithelium with Lotus tetragonologus a variable loss of binding has been found within carcinomas. Surprisingly, this bears no relationship to tumour differentiation; nor does it correlate with local lymph node metastasis. Ulex europeus I has a variable reactivity with carcinomas, as it does with normal breast epithelium, and this likewise has no correlation with differentiation. However, a relationship has been found between specific patterns of binding and axillary lymph node status. The value of using two lectins with the same major sugar specificity but having different binding affinitics is illustrated in this study, in which differences in behavioural characteristics have been identified between fucose‐containing glycoconjugates having minor structural variations.
Alterations in the biophysical properties of the human endothelial cell plasma membrane induced by a chemotactic tripeptide: Correlation with enhanced adherence of granulocytes
Authors: Kirkpatrick Charles J. , Melzner Ingo
Published: June 2005

Human umbilical vein endothelial cells in monolayer culture were used to study the effects of the chemotactic tripeptide, N‐formylmethionylleucylphenylalanine (FMLP), on structure and function of the endothelium. Endothelial cell morphology was unaffected by concentrations of 10−8−10−4 M. No effect on endothelial cell proliferative capacity, as measured by the DNA content of cultures, was seen at the FMLP concentrations studied (10−8−10−6 M). Using fluorescent molecular probes to investigate FMLP‐induced alterations in membrane structure, it was shown using the monomer–excimer method with pyrene decanoic acid that FMLP caused a marked restructuring of the plasma membrane. This took the form of a restriction of the surface available to the lipophile reporter molecules, probably caused by a molecular reorganization of the membrane protein component. Experiments with diphenylhexatriene indicated that FMLP did not make the plasma membrane of the endothelial cell more fluid. Concomitant with these changes in the physical properties of the membrane, an FMLP‐induced increase in granulocyte adherence to the endothelial cells was observed. A theoretical model is presented correlating granulocyte adherence with the lateral mobility of lipids in the endothelial cell membrane. The significance of the FMLP‐induced increase in granulocyte adherence to endothelial cells for the pathogenesis of sepsis is discussed.
Paranodal tumour in breast cancer: Extranodal extension versus vascular spread
Authors: Hartveit F.
Published: June 2005

A detailed histological study of the axillary nodes from 98 breast carcinoma patients, in which 5 and 10 year follow‐up data was available, has been undertaken. The presence of tumour cells in efferent vessels appears to be the only factor which is a marker of poor prognosis. Contrary to previous reports the direct infiltration of tumour cells through the nodal capsule into perinodal fat and connective tissue is of no prognostic significance. Distinction between site and type of tumour growth within nodes is thus essential for prognostic evaluation.
Cell surface characterization of the human osteoclast: Phenotypic relationship to other bone marrow‐derived cell types
Authors: Horton M. A. , Rimmer E. F. , Lewis D. , Pringle J. A. S. , Fuller K. , Chambers T. J.
Published: June 2005

Among the differentiated progeny of stem cells transplantable by bone marrow are osteoclasts, the multinucleate cells which are the major agents of bone resorption. Although the osteoclast is well characterized from a structural and functional standpoint, its development and origin are still far from clear. We have used monoclonal antibodies to investigate the interrelationship between osteoclasts and other haemopoietic cells in man. We have analysed the distribution of 19 granulocyte–monocyte antigens in eight reactivity clusters on the non‐neoplastic osteoclasts present within nine osteoclastomas (syn. giant cell tumours of bone) and a single example of aneurysmal bone cyst. We found that osteoclasts are antigenically effete, failing to express granulocyte‐monocyte, common leucocyte or other haemopoietic determinants; the only monocyte antigens detected on osteoclasts are My‐7 and two closely related specificities, MCS.2 and DüHL60.4, which are also expressed by tissues outside the haemopoietic system. Our findings, taken together with recent transplantation studies, cast further doubt on the view that osteoclasts are specialized bone‐resorbing macrophage‐derived giant cells, and support a hypothesis that they are the end product of fusion of a hitherto unidentified circulating mononuclear cell type, the preosteoclast, which constitutes a cell lineage separate from those originating from the conventional multipotential haemopoietic stem cell, although still of bone marrow origin.
The structure and interactions of Ca2+-ATPase
Authors: Martonosi Anthony N.
Published: June 2005

Electron crystallographic studies on membrane crystals of Ca2+-ATPase reveal different patterns of ATPase-ATPase interactions depending on enzyme conformation. Physiologically relevant changes in Ca2+ concentration and membrane potential affect these interactions. Ca2+ induced difference FTIR spectra of Ca2+-ATPase triggered by photolysis of caged Ca2+ are consistent with changes in secondary structure and carboxylate groups upon Ca2+ binding; the changes are reversed during ATP hydrolysis suggesting that a phosphorylated enzyme form of low Ca2+ affinity is the dominant intermediate during Ca2+ transport. A two-channel model of Ca2+ translocation is proposed involving the membrane-spanning helices M2–M5 and M4, M5, M6 and M8 respectively, with separate but interacting Ca2+ binding sites.
Lipid structure and Ca2+-ATPase function
Authors: Lee Anthony G. , Dalton Kate A. , Duggleby Richard C. , East J. Malcolm , Starling Anthony P.
Published: June 2005

Effects of lipid structure on the function of the Ca2+-ATPase of skeletal muscle of sarcoplasmic reticulum are reviewed. Binding of phospholipids to the ATPase shows little specificity. Phosphatidylcholines with short (C14) or long (C24) fatty acyl chains have marked effects on the activity of the ATPase, including a change in the stoichiometry of Ca binding. Low ATPase activity in gel phase lipid follows from low rate of phosphorylation. Phosphatidylinositol 4-phosphate increases ATPase activity by increasing the rate of dephosphorylation of the phosphorylated ATPase. Stimulation is not seen with other anionic phospholipids; phosphatidic acid decreases ATPase activity in a Mg2+-dependent manner.
The SERCA3-type of organellar Ca2+pumps
Authors: Wuytack Frank , Dode Leonard , Baba-Aissa Fouzia , Raeymaekers Luc
Published: June 2005

Of all the SERCA pumps, SERCA3 was the latest to be described and the least well known. Its primary structure deviates more than usual from the other members of the SERCA family. It is not known whether its remarkably low affinity for Ca2+ (K0.5 > 1μM) observed upon expression in the COS cell system occurs also in its normal cellular context. SERCA3 is particularly expressed at high levels in different types of blood cells and related cells like platelets, lymphocytes, mast cells and arterial endothelial cells. It is also found in cerebellar Purkinje neurons. The physiological significance of this expression pattern remains unknown.
Modulation of SERCA2 activity: Regulated splicing and interaction with phospholamban
Authors: Verboomen Hilde , Mertens Luc , Eggermont Jan , Wuytack Frank , Van Den Bosch Ludo
Published: June 2005

Ca2+-uptake into intracellular stores is mediated by the sarco/endoplasmic reticulum Ca2+ATPases (SERCAs). This review deals first with the gene structural and the characterization of the tissue-specific SERCA2 transcript processing. Secondly, the two different protein isoforms and their regulation are described. Finally, this review ends with a discussion on the possible physiological role of the SERCA2 isoform diversity.
Fluorometric study on conformational changes in the catalytic cycle of sarcoplasmic reticulum Ca2+-ATPase
Authors: Kanazawa Tohru , Suzuki Hiroshi , Daiho Takashi , Yamasaki Kazuo
Published: June 2005

Changes in the fluoresence of N-acetyl-N′-(5-sulfo-1-naphthyl)ethylenediamine (EDANS), being attached to Cys-674 of sarcoplasmic reticulum Ca2+-ATPase without affecting the catalytic activity, as well as changes in the intrinsic tryptophan fluorescence were followed throughout the catalytic cycle by the steady-state measurements and the stopped-flow spectrofluorometry. EDANS-fluorescence changes reflect conformational changes near the ATP binding site in the cytoplasmic domain, while tryptophan-fluorescence changes most probably reflect conformational changes in or near the transmembrane domain in which the Ca2+ binding sites are located. Formation of the phosphoenzyme intermediates (EP) was also followed by the continuous flow-rapid quenching method. The kinetic analysis of EDANS-fluorescence changes and EP formation revealed that, when ATP is added to the calcium-activated enzyme, conformational changes in the ATP binding site occur in three successive reaction steps; conformational change in the calcium enzyme substrate complex, formation of ADP-sensitive EP, and transition of ADP-sensitive EP to ADP-insensitive EP. In contrast, the ATP-induced tryptophan-fluorescence changes occur only in the latter two steps. Thus, we conclude that conformational changes in the ATP binding site in the cytoplasmic domain are transmitted to the Ca2+-binding sites in the transmembrane domain in these latter two steps.
Ca2+ binding and translocation by the sarcoplasmic reticulum ATPase: Functional and structural considerations
Authors: Inesi Giuseppe , Chen Li , Sumbilla Carlota , Lewis David , Kirtley Mary E.
Published: June 2005

Three experimental systems are described including sarcoplasmic reticulum (SR) vesicles, reconstituted proteoliposomes, and recombinant protein obtained by gene transfer and expression in foreign cells. It is shown that the Ca2+ ATPase of sarcoplasmic reticulum (SR) includes an extramembranous globular head which is connected through a stalk to a membrane bound region. Cooperative binding of two calcium ions occurs sequentially, within a channel formed by four clustered helices within the membrane bound region. Destabilization of the helical cluster is produced following enzyme phosphorylation by ATP at the catalytic site in the extramembranous region. The affinity and orientation of the Ca2+ binding site are thereby changed, permitting vectorial dissociation of bound Ca2+ against a concentration gradient. A long range linkage between phosphorylation and Ca2+ binding sites is provided by an intervening peptide segment that retains high homology in cation transport ATPases, and whose function is highly sensitive to mutational perturbations.
Use of thapsigargin to study Ca2+ homeostasis in cardiac cells
Authors: Rogers Terry B. , Inesi Giuseppe , Wade Robert , Lederer W. J.
Published: June 2005

Several reports have documented that thapsigargin is a potent inhibitor of the SR Ca2+ ATPase isolated from cardiac or skeletal muscle. We have characterized the specificity of this agent in intact rat cardiac myocytes using cells maintained in the whole cell voltage clamp configuration. We have shown that thapsigargin decreases the magnitude of the Ca2+ transient and the twitch by about 80% while it slows the decay rate for these responses. These changes were not accompanied by any alterations in sarcolemmal currents or in the trigger Ca2+ generated by the inward calcium current. Taken together these results reveal that the action of thapsigargin is restricted to the SR Ca2+ ATPase in intact cardiac myocytes. Furthermore, it is demonstrated unambiguously that SR intracellular Ca2+ stores are an absolute requirement for the development of contractile tension in rat heart myocytes. It is shown that thapsigargin is a valuable probe to examine the importance of SR pools of Ca2+ and the role of the Ca2+ ATPase in intact myocytes as well as in genetically altered heart cells.
Transmembrane Ca2+ gradient and function of membrane proteins
Authors: Yang F. Y. , Huang Y. G. , Tu Y. P.
Published: June 2005

This review will focus on the recent advance in the study of effect of transmembrane Ca2+ gradient on the function of membrane proteins. It consits of two parts: 1. Transmembrane Ca2+ gradient and sarcoplasmic reticulum Ca2+-ATPase; 2. Effect of transmembrane Ca2+ gradient on the components and coupling of cAMP signal transduction pathway. The results obtained indicate that a proper transmembrane Ca2+ gradient may play an important role in modulating the conformation and activity of SR Ca2+-ATPase and the function of membrane proteins involved in the cAMP signal transduction by mediating the physical state change of the membrane phospholipids.
Ligand-gated channel of the sarcoplasmic reticulum Ca2+ transport ATPase
Authors: Wolosker Herman , de Meis Leopolde
Published: June 2005

In resting muscle, cytoplasmic Ca2+ concentration is maintained at a low level by active Ca2+ transport mediated by the Ca2+ ATPase from sarcoplasmic reticulum. The region of the protein that contains the catalytic site faces the cytoplasmic side of the membrane, while the transmembrane helices form a channel-like structure that allows Ca2+ translocation across the membrane. When the coupling between the catalytic and transport domains is lost, the ATPase mediates Ca2+ efflux as a Ca2+ channel. The Ca2+ efflux through the ATPase channel is activated by different hydrophobic drugs and is arrested by ligands and substrates of the ATPase at physiological pH. At acid pH, the inhibitory effect of cations is no longer observed. It is concluded that the Ca2+ efflux through the ATPase may be sufficiently fast to support physiological Ca2+ oscillations in skeletal muscle, that occur mainly in conditions of intracellular acidosis.
Luminal calcium regulation of calcium release from sarcoplasmic reticulum
Authors: Hidalgo Cecilia , Donoso Paulina
Published: June 2005

This article discusses how changes in luminal calcium concentration affect calcium release rates from triad-enriched sarcoplasmic reticulum vesicles, as well as single channel opening probability of the ryanodine receptor/calcium release channels incorporated in bilayers. The possible participation of calsequestrin, or of other luminal proteins of sarcoplasmic reticulum in this regulation is addressed. A comparison with the regulation by luminal calcium of calcium release mediated by the inositol 1,4,5-trisphosphate receptor/calcium channel is presented as well.
Dihydropyridine receptor-ryanodine receptor interactions in skeletal muscle excitation-contraction coupling
Authors: Meissner Gerhard , Lu Xiangyang
Published: June 2005

Much recent progress has been made in our understanding of the mechanism of sarcoplasmic reticulum Ca2+ release in skeletal muscle. Vertebrate skeletal muscle excitation-contraction (E-C) coupling is thought to occur by a “mechanical coupling”� mechanism involving protein-protein interactions that lead to activation of the sarcoplasmic reticulum (SR) ryanodine receptor (RyR)/Ca2+ release channel by the voltage-sensing transverse (T−) tubule dihydropyridine receptor (DHPR)/Ca2+ channel. In a subsequent step, the released Ca2+ amplify SR Ca2+ release by activating release channels that are not linked to the DHPR. Experiments with mutant muscle cells have indicated that skeletal muscle specific DHPR and RyR isoforms are required for skeletal muscle E-C coupling. A direct functional and structural interaction between a DHPR-derived peptide and the RyR has been described. The interaction between the DHPR and RyR may be stabilized by other proteins such as triadin (a SR junctional protein) and modulated by phosphorylation of the DHPR.
Fetal and neonatal bile acid synthesis and metabolism — Clinical implications
Authors: Balistreri W. F.
Published: June 2005

It has become apparent that with sophisticated technology we are now able to recognize defective bile acid metabolism in a wide variety of disease states. Recognition of specific aberrations, such as inborn errors in bile acid metabolism manifesting as neonatal cholestasis, offers new opportunities for therapeutic intervention. Future studies should determine the incidence of inborn errors in patients with enigmatic and unexplained liver diseases such as idiopathic neonatal hepatitis.
Clinical approach to inherited metabolic diseases in the neonatal period: A 20‐year survey
Authors: Saudubray J. M. , Ogier H. , Bonnefont J. P. , Munnich A. , Lombes A. , Hervé F. , Mitchel G. , Poll Thé B. , Specola N. , Parvy P. , Bardet J. , Rabier D. , Coudé M. , Charpentier C. , Frézal J.
Published: June 2005

Every newborn with unexplained neurological deterioration, ketosis, metabolic acidosis or hypoglycaemia should be suspected of having an inherited error of intermediary metabolism. Many of these conditions can be diagnosed clinically with the aid of simple laboratory investigations. Since a substantial number of these diseases respond well to treatment but may otherwise be fatal, and in order to assure adequate prenatal diagnosis in subsequent pregnancies, a high index of suspicion and rapid diagnosis are necessary in the face of the clinical presentations described. According to three major clinical presentations observed in 218 neonates with inborn errors of intermediary metabolism (neurological distress ‘intoxication’ type, neurological distress ‘energy deficiency’ type and hypoglycaemia with liver dysfunction) and according to the proper use of few laboratory investigations, we propose a method of diagnosis which groups these children into five categories. Initial therapy, and sophisticated investigations can be planned on the basis of this grouping.
A clinician's view of the mass screening of the newborn for inherited diseases: Current practice and future considerations
Authors: Sardharwalla I. B. , Wraith J. E.
Published: June 2005

The case for or against mass screening for inherited diseases is discussed. There is universal acceptance for mass screening for phenylketonuria and congenital hypothyroidism. The case for mass screening for galactosaemia and for maple syrup urine disease is not very strong; they could be considered under the heading of ‘urgent screening of the sick newborn’. It is difficult to find good arguments for mass screening for congenital adrenal hyperplasia. For screening for glucose‐6‐phosphate dehydrogenase deficiency and sickle cell disease, the established criteria for mass screening do not apply. A simple tool for early detection is now available and the population afflicted with a mutant gene which causes major health problems should receive special attention from its government. It is too early to offer any comment about cystic fibrosis screening; further developments must be awaited.
Prospects for gene therapy now and in the future
Authors: Akhurst R. J.
Published: June 2005

Advances in gene technology and cell biology have supplied the means to undertake human gene therapy in the near future. Techniques have been developed for the efficient introduction of gene sequences into the pluripotential stem cells of the haematopoietic system and our increased understanding of gene‐regulatory mechanisms should allow therapeutic gene expression levels to be obtained.

Gene therapy should, at present, be termed gene supplementation since it will involve the addition of corrective genes to the host cell genome. It may only be used to treat recessively inherited disorders. Prospects for the future include the use of homologous recombination to correct or replace defective genes, allowing the treatment of dominantly inherited diseases.
Partial lyonisation of steroid sulphatase gene in single hair roots
Authors: Willems P. J. , de Bruijn H. W. A. , Groenhuis A. , Mooyaart B. R. , Berger R.
Published: June 2005

Steroid sulphatase (STS) activity was measured in single hair roots from 9 patients with X‐linked ichthyosis, 7 obligate heterozygotes, 12 male controls and 12 female controls. STS activity was deficient (<0.3 pmol dehydroepian‐drosterone (DHEA) in 5 h per hair root) in the patients as compared to male controls (4.6±1.8 pmol DHEA in 5 h per hair root). All single hair roots from obligate heterozygotes showed STS activity (3.0±0.9 pmol DHEA in 5 h per hair root). Female controls had a significantly higher STS activity (6.0±1.8 pmol DHEA in 5 h per hair root) than male controls (p<0.05) and heterozygotes (p<0.001). Both the female‐male sex ratio of control STS activity (1.3) and the absence of STS‐deficient hair roots in heterozygotes confirm previous evidence for incomplete lyonisation of the X‐linked steroid sulphatase gene.
Regional distribution of the phenylalanine‐sensitive ATP‐sulphurylase in brain
Authors: Matsuo K. , Hommes F.
Published: June 2005

The regional distribution of the phenylalanine‐sensitive ATP‐sulphurylase in fetal calf brain coincides with demyelinated lesions observed in the central nervous systems of untreated PKU patients. This would be expected if this species of ATP‐sulphurylase played a role in the pathogenesis of brain dysfunction in the untreated or poorly controlled phenylketonuria patient.
HPRT‐Deficiency associated with normal PRPP concentration and APRT activity
Authors: Gordon R. B. , Keough D. T. , Emmerson B. T.
Published: June 2005

Deficiencies of HPRT are usually associated with increased concentrations of PRPP and increased levels of APRT activity in erythrocytes. We report the case of a male with a partial deficiency of HPRT in whom these two parameters were normal. The clinical features of this patient were those associated with severe hyperuricaemia and gout. Studies of intact erythrocytes showed rates of incorporation of [14C]hypoxanthine and of [14C]adenine into purine nucleotides which were almost indistinguishable from normal. However, HPRT activity in erythrocyte lysates was only 9% of normal. In cell extracts of cultured lymphoblasts, the HPRT activity was 20% of control values and the APRT activity was normal. The PRPP concentration and the rate ofde novo purine synthesis in cultured lymphoblasts were both intermediate between controls and lymphoblasts from patients with the Lesch‐Nyhan syndrome.
Phosphorylase b kinase deficiency in man: a review
Authors: van den Berg I. E. T. , Berger R.
Published: June 2005

Phosphorylase b kinase is involved in the activation of glycogen phosphorylase and is thus involved in the breakdown of glycogen. The enzyme exists as several tissue specific isoenzymes of which the muscle enzyme (rabbit) has been most characterized. It is a multimeric protein composed of four subunits, α, β, γ and δ. The four subunits are coded on different chromosomes, the α, β and γ subunit genes being on the X, 16 and 17 chromsomes respectively. The δ subunit is a calmodulin and confers calcium sensitivity on phosphorylase b kinase. Tissue specificity of the enzyme is conferred, at least in some cases, by variation in the γ subunit.

Seven different clinical types of phosphorylase b kinase deficiency have been described. The most common type is X‐linked and affects the liver only; other types affect liver, muscle and liver, muscle or heart and have an autosomal recessive mode of inheritance, while in some types the mode of inheritance is not clear. Diagnosis based on the study of erythrocytes or leukocytes can be misleading due to the tissue specific nature of the enzyme, and liver or muscle biopsies may be required.
A preliminary report of the collaborative study of maternal phenylketonuria in the United States and Canada
Authors: Koch R. , Hanley W. , Levy H. , Matalon R. , Rouse B. , Cruz F. Dela , Azen C. , Friedman E. Gross
Published: June 2005

The Maternal Phenylketonuria Collaborative Study (MPKUCS), encompassing all the United States and provinces of Canada, is a prospective, longitudinal investigation designed to ascertain the efficacy of phenylalanine‐restricted therapy in protecting the fetus from high maternal phenylalanine concentrations in women with hyperphenylalaninaemia.

Preliminary findings are reported for 147 pregnancies for whom the recommended therapeutic range of blood phenylalanine was 120–360 µmol/L. Sixty‐three pregnancies had complete data for analysis. Dietary control was attempted prior to conception in 10 out of 63 women. Significant negative correlations were noted in length, weight and head circumference and blood phenylalanine concentrations during pregnancy. Average reported phenylalanine levels by trimester for 63 hyperphenylalaninaemic pregnancies resulting in live births revealed that no group requiring treatment achieved levels below 360 µmol/L until the third trimester. Median birth measurement percentiles revealed that all groups studied generally had smaller head size compared with birth length and weight. Those started on diet after the first trimester achieved a head circumference below the 10th percentile.

The implication of small head circumference for subsequent intellectual development is unclear at this time. Furthermore, the study must evaluate more offspring of women having optimal preconception and pregnancy restriction of phenylalanine.
A retrospective study of a patient with homozygous form of acute intermittent porphyria
Authors: Beukeveld G. J. J. , Wolthers B. G. , Nordmann Y. , Deybach J. C. , Grandchamp B. , Wadman S. K.
Published: June 2005

In 1964 a child with an exceptional form of porphyria was described; she excreted persistently excessive amounts of delta‐aminolaevulinic acid, porphobilinogen and uroporphyrin in her urine from early childhood. The biochemical profile resembled that of acute intermittent porphyria (AIP). The child died at the age of 8 years. Reinvestigation of some urine samples by HPLC revealed differences in comparison with urines of other patients with AIP. The clinical picture characterized by porencephaly and severe retardation in development was completely different from that of AIP. Her mother suffered from AIP but the father never had attacks. Investigations on blood and urine samples of the father showed that he also was affected. Due to the early onset in the index patient, its persistent character, and the fact that both parents are affected we postulate retrospectively to have diagnosed a case of homozygous or a double heterozygous AIP, hitherto undescribed.
Impaired sulphated glycosaminoglycan metabolism in a patient with GM‐2 gangliosidosis (Tay‐Sachs disease)
Authors: Toma L. , Pinto W. , Rodrigues V. C. , Dietrich C. P. , Nader H. B.
Published: June 2005

An abnormal urinary excretion of sulphated glycosaminoglycans in a patient with GM‐2 gangliosidosis (Tay‐Sachs disease) is described. Besides the accumulation of GM‐2 ganglioside in liver and lack of hexosaminidase A, the patient shows an abnormal urinary excretion of an iduronic acid‐rich low molecular weight heparan sulphate. Also, no dermatan sulphate could be detected in the urine, whereas this compound was the main sulphated glycosaminoglycan in the liver of the patient. Heparan sulphate was the main glycosaminoglycan of normal liver. The total amount of sulphated glycosaminoglycans in the urine and liver of the patient did not differ significantly from the amounts found in the liver and urine of normal subjects. Several plasma glycosidases have been assayed and the activities did not differ significantly from the values obtained for the plasma of normal subjects.
Structure and function of the mitochondrial genome
Authors: Clayton D. A.
Published: June 2005

It is now clear that molecular defects in human mitochondrial DNA play a significant role in human disease. Mitochondrial DNA mutations range from single base changes in the 16.5 kilobase‐pair genome up to large deletions and rearrangements. Independently of the actual cause of a given mutation, it is possible to predict at least some of the consequences of changes in mitochondrial DNA sequence. This paper reviews our overall understanding of the mode and mechanism of mitochondrial DNA replication and transcription and how this relates to mitochondrial gene expression. This provides a background to anticipate the nature and extent of mitochondrial DNA sequence changes that might be of physiological consequence.
Abnormalities of human sex determination
Authors: Ferguson‐Smith M. A.
Published: June 2005

Cytogenetic and molecular studies in patients with abnormalities of sex determination have been the key to the isolation and investigation of candidates for the primary testis determining factor (TDF). A gene, SRY, isolated from the sex determining region of the Y chromosome within 5 kilobases of the pairing segment boundary, has been characterized recently which fulfils the expectations of TDF. It is expressed in the embryonic gonads at the critical time of differentiation; it is highly conserved among mammals; it has the structure of a transcription regulator; and mutations within its conserved domain are found in 10% of sex‐reversed XY females. The murine homologue of this gene has been shown to cause sex reversal in XX embryos following injection of a 14 kb DNA fragment containing SRY into fertilized eggs. However, most XX true hermaphrodites and a proportion of XX sex‐reversed males lack SRY despite the presence of testicular differentiation. It is postulated that the constitutive activation of an X‐linked gene, TDF‐2, normally regulated by SRY, is responsible for male differentiation in these cases. The female phenotype of XY individuals with duplications of Xp may be the result or deletion of disruption of TDF‐2.
Prenatal diagnosis of Hurler disease by analysis of α‐iduronidase in chorionic villi
Authors: Young E. P.
Published: June 2005

Twenty‐four pregnancies at risk for Hurler disease (MPS I) were monitored by measurement of α‐iduronidase in chorionic villi. Adequate samples were obtained for direct assay of the villi in 22 pregnancies. Five were found to be affected and the pregnancies were terminated. In another pregnancy an equivocal result was obtained on direct assay but analysis of the cultured chorionic cells showed the fetus to be affected. In one pregnancy where an exceptionally small biopsy was obtained, direct assay indicated the fetus to be unaffected. Following amniocentesis this result was shown to be incorrect. These results confirm that, provided an adequate sample is obtained, an accurate diagnosis can be made by direct assay of chorionic villi in pregnancies at risk for Hurler disease.
Prospective versus clinical diagnosis and therapy of acute neonatal hyperammonaemia in two sisters with carbamyl phosphate synthetase deficiency
Authors: Tuchman M. , Mauer S. M. , Holzknecht R. A. , Summar M. L. , Vnencak‐Jones C. L.
Published: June 2005

Two female siblings were treated for acute neonatal hyperammonaemia due to complete carbamyl phosphate synthetase I deficiency. The first child was detected clinically at 65 hours of age and therapy started at 79 hours. The second child was followed from birth and therapy started at 5 hours of age. The extrapolated rate of increase of blood ammonia, in the first hours of life before therapy started, was 19 µmol L−1 h−1 in both babies. Peak blood ammonia level was 2235 µmol/L in the first (clinically detected) child and 271 µmol/L in the second (prospectively followed) child. The second child became symptomatic at 3 hours of age when blood ammonia level was as low as 90 µmol/L, whereas blood ammonia levels above 100 µmol/L caused no symptoms during recovery. The child detected clinically required haemodialysis and peritoneal dialysis to treat the hyperammonaemia. In the prospectively treated child, early therapy with intravenous sodium benzoate and sodium phenylacetate slowed the rate of increase in blood ammonia level, but this therapy did not prevent the need for peritoneal dialysis.
Neuropathological and clinical correlations in hurler disease
Authors: Watts R. W. E. , Spellacy E. , Hume Adams J.
Published: June 2005

We report studies on two patients (1 and 2) with Hurler disease. They both had all of the non‐neurological features of Hurler disease to a similar and extreme degree and similar signs of brain damage on computed tomography. However, intellectual function was unusually well‐preserved in patient 1, but seriously and typically impaired in patient 2. The reason for this discrepancy has been investigated by reference to the neuropathological findings, the results of α‐l‐iduronidase assays using different substrates and comparisons to other cases (patients 3 and 4). We suggest that patient 1 is an unusual variant of the disease who may have had a very low residual α‐l‐iduronidase activity in neuronal cells only, and that this could not be demonstrated by either enzyme assays on whole brain using the 4‐methylumbelliferyliduronide substrate (Crowet al., 1983) or in studies on fibroblast lysates using a radioactive disaccharide substrate.
Incidence of disorders tested by systematic screening: Confidence limits and comparison of programmes
Authors: Bachmann C. , Colombo J. ‐P.
Published: June 2005

Using the data from screening done in Switzerland since 1965, we showed that the probability of finding cases with increased phenylalanine or leucine concentrations is compatible with a Poisson distribution. If there are no trends present from year to year and if the Poisson distribution is an accurate fit further statistical treatment is possible. The advantage of using the Poisson distribution is exemplified by the calculation of confidence limits for the incidence of hyperphenylalaninaemia, maple syrup urine disease and hypothyroidism. Furthermore, statistical comparisons between different screening programs are easily done.
Pregnancy and argininosuccinic aciduria
Authors: Worthington S. , Christodoulou J. , Wilcken B. , Peat B.
Published: June 2005

We present the outcome of a pregnancy in a woman with mild argininosuccinic lyase deficiency to add to the collective experience of the maternal and fetal effects of urea cycle defects. In females affected with argininosuccinic lyase deficiency, careful clinical and biochemical monitoring of pregnancy will minimize the risk of metabolic decompensation in the perinatal period. Furthermore, it would appear that argininosuccinate is not teratogenic to the development of the human fetus.
Major hyperpipecolataemia in a normal adult
Authors: Vallat C. , Denis S. , Bellet H. , Jakobs C. , Wanders R. J. A. , Mion H.
Published: June 2005

We describe the fortuitous discovery of a 44‐year‐old man with a very high hyperpipecolataemia (250 µmol/L; normal <2.5). This patient has none of the clinical features seen in peroxisomal diseases, he is a strictly normal intelligent adult. A stereochemical study of this pipecolic acid was performed usingd‐amino acid oxidase, and identified it asl‐pipecolic acid. We suggest that isolatedl‐hyperpipecolataemia may be a benign trait.
First trimester prenatal diagnosis of metachromatic leukodystrophy on chorionic villi by ‘Immunoprecipitation—electrophoresis’
Authors: Poenaru L. , Castelnau L. , Besançon A‐M. , Nicolesco H. , Akli S. , Theophil D.
Published: June 2005

Prenatal diagnosis of metachromatic leukodystrophy (MLD) due to arylsulphatase A (ASA) deficiency can be performed by amniocentesis with the disadvantage of a late pregnancy termination.

Whether chorionic villi (CV) obtained by trophoblast biopsy during the first trimester of pregnancy can be useful for diagnosis depends on the reliability of results. The complexity of arylsulphatase expression in CV and the existence of several isozymes make diagnosis difficult. However, the use of an anti‐ASA antiserum enabled us to discriminate between ASA and a comigrating contaminant isozyme, and the antigen‐antibody (Ag‐Ab) complex gave better evidence of the presence or absence of ASA after enhancement of activity with 4‐methylumbelliferyl sulphate (4‐MUS).

We propose that immunoprecipitation followed by electrophoresis could be a valuable method of MLD prenatal diagnosis on chorionic villi.
Prolidase deficiency: A patient without hydroxyproline‐containing iminodipeptides in urine
Authors: Wysocki S. J. , Hahnel R. , Mahoney T. , Wilson R. G. , Panegyres P. K.
Published: June 2005

A 17‐year‐old girl was shown to have prolidase deficiency on the basis of the presence of large amounts of proline‐containing dipeptides in urine and an almost complete absence of prolidase in plasma and erythrocytes. Unlike most earlier cases of this genetic defect our patient did not excrete hydroxyproline‐containing dipeptides in her urine.
Pristanic acid does not accumulate in peroxisomal acyl‐CoA oxidase deficiency: Evidence for a Distinct peroxisomal pristanyl‐CoA oxidase
Authors: ten Brink H. J. , Poll‐The B. T. , Saudubray J. M. , Wanders R. J. A. , Jakobs C.
Published: June 2005

The concentration of pristanic acid was measured in plasma from a patient with an isolated peroxisomal very long chain fatty acid (VLCFA) acyl‐CoA oxidase deficiency, a defect in peroxisomalβ‐oxidation resulting in accumulation of VLCFA in plasma and tissues. Although peroxisomes are believed to be involved in pristanic acidβ‐oxidation, the pristanic acid level in the patient's plasma was within the control range. This finding provides evidence for the existence of a pristanyl‐CoA oxidase distinct from the specific trihydroxycholestanoyl‐CoA and VLCFA acyl‐CoA oxidases.