bluuebunny/crossref_metadata_2025 huggingface dataset:
A French Translation of the Obsessive-Compulsive Drinking Scale for Craving in Alcohol-Dependent Patients: A Validation Study in Belgium, France, and Switzerland
Authors: Ansseau M. , Besson J. , Lejoyeux M. , Pinto E. , Landry U. , Cornes M. , Deckers F. , Potgieter A. , Ades J.
Published: April 2003

The Obsessive-Compulsive Drinking Scale (OCDS) is an instrument developed to measure cognitive aspects of alcohol craving. The aim of this study was to validate the French translation of the OCDS according to the international methodology as published by Hunt and coworkers (see text), including forward-backward translations, patient interviews (9 patients), patient’s perception of acceptability (15 patients), and final validation within a treatment program (50 patients). All 74 patients were native French-speaking alcohol-dependent patients from Belgium, France, and Switzerland. The derived aggregated total (TOT) score and both subscores corresponding to the obsessive (OB) and compulsive (CP) dimensions were shown to be asymptomatically normal. Good internal consistencies were found, with Cronbach α: TOT = 0.88; OB = 0.82; CP = 0.79. The test-retest procedure was used to examine intrarater reliability (r = 0.81). The construct validity was examined with linear correlation of the two main components: r(OB, CP) = 0.62; r(OB, TOT) = 0.86; r(CP, TOT) = 0.92. Principal-components analysis revealed two main factors: the first factor representing the total scale score, while the obsessive and compulsive subscale scores were distributed along factor two. The translated scale seems to be psychometrically as valid as the original English scale and confirms the psychometric properties of the OCDS.
DQA1*Arg52, DQB1*nonAsp57, and DRB1*04 Genotypes in Chinese Children with Insulin-Dependent Diabetes mellitus
Authors: Huang Fu-Yuan , Lee Yann-Jinn , Lo Fu-Sung , Wang Chao-Hung , Lin Shuan-Pei , Hsu Chyong-Hsin , Kao Hsin-An , Tsan Kun-Wu , Chang Jan-Gowth
Published: April 2003

Ethnic comparisons are extremely important and useful for studying the HLA component involved in insulin-dependent diabetes mellitus (IDDM) predisposition. To date there have been only a few reports on the association of HLA loci and IDDM in Chinese. We report here a study on DQA1*Arg52, DQB1*nonAsp57, and DRB1*04 in IDDM children and control adults among Han Chinese living in Taiwan. One hundred and fourteen unrelated children (62 boys) with IDDM were studied. Their ages at diagnosis were between 0.3 and 15.0 years (6.8 ± 3.6 years). The control population consisted of 120 randomly selected normal adults. DQA1*Arg52(+/+), DQB1*nonAsp57(+/+), and DRB1*04(+/–) were associated with IDDM (RR = 11.50, 2.21, and 2.82; p = 1.11 × 10–15, 2.84 × 10–3, and 1.98 × 10–4, respectively). DQA1*Arg52, DQB1*nonAsp57, and DRB1*04 conferred risks for IDDM (RR = 12.79, 7.11, and 2.83; pc = 8.22 × 10–4, 5.35 × 10–3, and 5.68 × 10–4, respectively). Combinations of DQA1*Arg52 and DRB1*04 conferred the highest risk for IDDM (RR = 19.64, pc = 5.4 × 10–5). DQA1*Arg52 was associated with IDDM in subjects with DQB1*nonAsp57+ (RR = 14.87, pc = 2.41 × 10–4) and DQB1*nonAsp57 was also associated with IDDM in subjects with DQA1*Arg52+ (RR = 8.41, pc = 1.54 × 10–3), suggesting that DQA1*Arg52 and DQB1*nonAsp57 are interacting. This study demonstrates that DQA1*Arg52, DQB1*nonAsp57, and DRB1*04 confer susceptibility for IDDM to Chinese children. A combination of DQA1*Arg52 and DRB1*04 confers the highest risk and it is suggested that a susceptibility gene might be situated between DQA1*Arg52 and DRB1*04 or both are synergistic. There is an interaction between DQA1*Arg52 and DQB1*nonAsp57 and homozygosity for DQA1*Arg52/DQB1*nonAsp57, which encodes four susceptibility DQ heterodimers, confers a high risk.
Seasonal Variations in T-Lymphocyte Response to Grass Pollen Allergens from Pollen-Allergic Patients and Healthy Controls
Authors: Jepsen K.F. , Nielsen L. , Olsen O.T. , Bendtzen K. , Poulsen L.K.
Published: April 2003

Previous studies have demonstrated the presence of grass pollen-specific T cells in grass pollen-allergic patients (GPA) as well as nonallergic subjects (NA). In order to elucidate a possible seasonal variation in proliferation and cytokine production of peripheral blood mononuclear cells (PBMC), PBMC from 13 GPA and 11 NA were stimulated with extracts of Phleum pratense and tetanus toxoid before and during two grass pollen seasons. IL-4, IL-5 and interferon-γ were determined by ELISAs. PBMC from GPA demonstrated a decreased proliferative response to grass pollen allergens during the pollen season as compared to NA (p < 0.05), but no difference was found in the response to tetanus toxoid. Cells from GPA produced higher amounts of IL-4 and IL-5 than NA (p < 0.05) and the seasonal variation in GPA proliferation was paralleled by the grass pollen-induced production of both IL-4 and IL-5 (p < 0.05). We conclude that during the grass pollen season PBMC from GPA have a reduced ability to proliferate and to produce Th2-type cytokines. This may be due to seasonal migration of the grass pollen-specific T cells from the blood to the tissues of primary allergen exposure.
A History of Complement Genetics
Authors: Alper Chester A.
Published: April 2003

After a brief review of the history of the complement system from the original observations in the 1880s, the history of complement genetics is outlined, beginning in 1919 with the recognition of the complement-deficient guinea pig. A highly personal view is then presented of the early days of the discovery of human complement structural gene polymorphisms and deficiencies, particularly those of C3, BF and factor I. Finally, some recent work based on features in the DNA of the MHC-encoded complement genes is outlined that sheds some light on their evolution.
Molecular Genetics of the Human MHC Complement Gene Cluster
Authors: Yu C. Yung
Published: April 2003

The human major histocompatibility complex (MHC) complement gene cluster (MCGC) is a highly variable region that is characterized by polymorphisms, variations in gene size and gene number, and associations with diseases. Deficiencies in complement C2 are either due to abolition of C2 protein synthesis by mini-deletions that caused frameshift mutations, or blocked secretion of the C2 protein by single amino acid substitutions. One, two or three C4 genes may be present in a human MCGC haplotype and these genes may code for C4A, C4B, or both. Deficiencies of C4A or C4B proteins are attributed to the expression of identical C4 isotypes or allotypes from the C4 loci, the absence or deletion of a C4 gene, 2-bp insertion at exon 29 or 1-bp deletion at exon 20 that caused frameshift mutations. The C4 genes are either 21 or 14.6 kb in size due to the presence of endogenous retrovirus HERV-K(C4) in the intron 9 of long C4 genes. A deletion or duplication of a C4 gene is always accompanied by its neighboring genes, RP at the 5′ region, and CYP21 and TNX at the 3′ region. These four genes form a genetic unit termed the RCCX module. In an RCCX bimodular structure, the pseudogene CYP21A, and partially duplicated gene segments TNXA and RP2 are present between the two C4 loci. The RCCX modular variations in gene number and gene size contributed to unequal crossovers and exchanges of polymorphic sequences/mutations, resulting in the homogenization of C4 polymorphisms and acquisitions of deleterious mutations in RP1, C4A, C4B, CYP21B and TNXB genes. RD, SKI2W, DOM3Z and RP1 are the four novel genes found between Bf and C4. RD and Ski2w proteins may be related to RNA splicing, RNA turnover and regulation of translation. The functions of Dom3z and RP1 are being investigated. The complete genomic DNA sequence between C2 and TNX is now available. This should facilitate a complete documentation of polymorphisms, mutations and disease associations for the MCGC.
The Human Immunoglobulin Heavy Variable Genes
Authors: Pallarès Nathalie , Lefebvre Sophie , Contet Valérie , Matsuda Fumihiko , Lefranc Marie-Paule
Published: April 2003

‘Human Immunoglobulin Heavy Variable Genes’, the fourth report of the ‘IMGT Locus on Focus’ section, comprises five tables entitled: (1) ‘Number of human germline IGHV genes at 14q32.33 and potential repertoire’; (2) ‘Human germline IGHV genes at 14q32.33’; (3) ‘Human IGHV orphons on chromosome 15 (15q11.2)’; (4) ‘Human IGHV orphons on chromosome 16 (16p11.2)’, and (5) ‘Human IGHV allele table’. These tables are available at the IMGT Marie-Paule page from IMGT, the international ImMunoGeneTics database (http://imgt.cnusc.fr:8104) created by Marie-Paule Lefranc, Université Montpellier II, CNRS, France.
Immunoglobulin Allotypes among the Bicolanos of Sorsogon Province, Luzon, Philippines: Implications of Phenotypes for Filariasis
Authors: Kron Michael A. , Ramirez Bernadette , Belizario Jr. Vicente , Pandey Janardan P.
Published: April 2003

Immunoglobulin (Ig) allotypes are polymorphic genetic systems that show distinct racial arrays, thus making them powerful tools for studies of genetic admixture and biological relationships. In the province of Sorsogon, southern Luzon, Philippines, allotyping was completed on 252 persons residing in two neighboring villages. The people demonstrated 14 GM and 3 KM phenotypes. The frequency of homozygous KM3, KM1 and heterozygous KM1,3 was identical in these villages; however, half of the GM phenotypes present in one village were significantly less frequent in the other village. The frequency of KM and GM haplotypes was different from those reported in Filipino aboriginal groups, but similar to a population on Samar Island, the only other Filipino group for which Ig allotype data exist. Variability in the prevalence of parasitic disease such as lymphatic filariasis may in part reflect differences in genetic susceptibility, resulting from allotypic heterogeneity between villages.
The Human Immunoglobulin Heavy Diversity (IGHD) and Joining (IGHJ) Segments
Authors: Ruiz Manuel , Pallarès Nathalie , Contet Valérie , Barbié Valérie , Lefranc Marie-Paule
Published: April 2003

The ‘Human Immunoglobulin Heavy Diversity (IGHD) and Joining (IGHJ) segments’, fifth report of the ‘IMGT Locus on Focus’ section, comprises six tables entitled: (1) ‘Human germline IGHD segments at 14q32.33’; (2) ‘Human IGHD alleles’; (3) ‘Human germline IGHJ segments at 14q32.33’; (4) ‘Human IGHJ alleles’; (5) ‘Human germline IGHD orphons on chromosome 15 (15q11.2)’; (6) ‘Correspondence between the different human IGHD nomenclatures’, and two figures: (1) ‘Protein display of human IGH D-REGIONs’; (2) ‘Protein display of human IGH J-REGIONs’. These tables and figures are available at the IMGT Marie-Paule page from IMGT, the international ImMunoGeneTics database (http://imgt.cnusc.fr:8104) created by Marie-Paule Lefranc, Université Montpellier II, CNRS, France.
Superantigens Are Presented by and Activate Thymocytes from Infants
Authors: Thulesen S. , Jørgensen A. , Gerwien J. , Dohlsten M. , Holst Nissen M. , Ødum N. , Röpke C.
Published: April 2003

A high percentage of human fetal and postnatal thymocytes express MHC class II molecules. This raises the possibility that human thymocytes in early life are able to present peptides to other immature T cells and thereby initiate thymic selection of these cells. Here we address this question by exposing newly harvested infant thymocytes to superantigen (Sag) which binds to the T-cell receptor and to MHC class II chains outside the peptide binding groove. The results show that the thymocytes are able to present Sag and to be activated to proliferation as well as apoptosis by Sag presented by other thymocytes. The absence of responses to Sag with mutations in class II binding sites showed that class II molecules were necessary for the responses, and very low expression of class II molecules on CD4–8– cells indicates that the demonstrated T-cell/T-cell interactions are confined to T-cell receptor-positive CD4+8+, CD4+8–, and CD4–8+ cells. These latter subsets were shown to be able to present Sag to each other. These findings suggest that class II+ thymocytes may participate in the selection of self-restricted T cells during a critical period in the shaping of the human immune system.
Lack of Association between CD28/CTLA-4 Gene Polymorphisms and Atopic Asthma in the Japanese Population
Authors: Nakao Futoshi , Ihara Kenji , Ahmed Saifuddin , Sasaki Yuka , Kusuhara Koichi , Takabayashi Akira , Nishima Sankei , Hara Toshiro
Published: April 2003

Atopic asthma occurs in genetically susceptible individuals in the presence of environmental factors. Recently, the costimulation signal from CD80-CD86 to CD28/CTLA-4 has been suggested to play an important role in the development of atopic asthma. In the present study, we analyzed three polymorphic regions within the CTLA-4 gene, an A/G substitution in exon 1 position 49, a C/T base exchange in the promoter position –318 and an (AT)n repeat polymorphism in the 3′-untranslated region of exon 4, and a CD28 gene polymorphism with a T/C substitution in intron 3 position +17 in 120 patients with atopic asthma and 200 normal controls. The polymorphism frequencies of CTLA-4/CD28 genes in patients did not differ from those in normal controls. Thus, the present study was unable to reveal any association between CTLA-4/CD28 gene polymorphisms and atopic asthma in the Japanese population.
Evidence for TGF-β-Mediated ‘Defense’ of the Glomerulus: A Blackguard Molecule Rehabilitated?
Authors: Kitamura Masanori , Fine Leon G.
Published: April 2003

Transforming growth factor beta (TGF-β) has been regarded as a ‘blackguard molecule’ that induces glomerular diseases. During the process of glomerulonephritis, upregulated TGF-β stimulates the production of extracellular matrix and inhibits its degradation, leading to excessive matrix deposition. On the other hand, TGF-β has the potential to be anti-inflammatory via inhibition of mitogenesis and production of inflammatory mediators by glomerular cells. This molecule strongly inactivates infiltrating cells, especially macrophages, which play a pivotal role in the generation of glomerular injury. The aim of this article is to summarize the potentially beneficial action of TGF-β in the glomerulus and to address its ‘bright side’ in glomerular inflammation.
Interleukin 10: A Logical Candidate for Suppressing Glomerular Inflammation?
Authors: Baud Laurent , Fouqueray Bruno , Suberville Sidonie , Doublier Sophie
Published: April 2003

Inflammatory processes within the glomerulus are switched off by the local generation of anti-inflammatory mediators. These mediators include eicosanoids (e.g., lipoxins), anti-inflammatory cytokines (interleukins 4 and 13), antagonists of proinflammatory cytokines (interleukin 1 receptor antagonist), neuropoietic cytokines (leukemia inhibitory factor and interleukin 6), as well as deactivators of inflammatory macrophages (transforming growth factor beta and interleukin 10). They limit the effects of proinflammatory mediators by inhibiting their production, stability, or function. Recent attempts to reduce inflammatory lesions in experimental glomerulonephritis have focused on upregulating the expression of these anti-inflammatory mediators by using protein or gene transfer. In particular administration of interleukin 4, interleukin 1 receptor antagonist, leukemia inhibitory factor, or interleukin 10 has been shown to be effective in the treatment of nephrotoxic nephritis. Of all the mediators already tested, interleukin 10 has the greatest potential because of its strong anti-inflammatory effects and weak adverse effects.
Low Catalase Activity in Rats with Ureteral Ligation: Relation to Lipid Peroxidation
Authors: Cvetković Tatjana , Vlahović Predrag , Pavlović Dušica , Kocić Gordana , Jevtović Tatjana , Djordjević Vidosava B.
Published: April 2003

Progression of some renal diseases is characterized by generation of reactive oxygen metabolites that are also involved in the pathophysiology of obstructive nephropathy. Catalase activity and lipid peroxidation were investigated in rats with unilaterally (UUL) and bilaterally ligated ureters (BUL). Forty-eight hours after ligation, the animals were sacrificed, and enzyme activity as well as the malondialdehyde (MDA) concentration were measured in the plasma, kidneys and livers. The activity of catalase was significantly reduced in the plasma of the BUL rats and in the kidneys of both investigated groups. In the liver, catalase activity was decreased only in the BUL group. The MDA concentration in the plasma and kidneys of the BUL rats was significantly increased while in the liver it remained unchanged. These results suggest that lipid peroxidation in the induced uremic state could be responsible for catalase inactivation.
Antisense Oligonucleotides
Authors: Kashihara Naoki , Maeshima Yohei , Makino Hirofumi
Published: April 2003

Antisense technology was developed to inhibit gene expression by utilizing an oligonucleotide complementary to the mRNA which encodes the target gene. There are a few possible mechanisms for the inhibitory effects of antisense oligonucleotides. Among them, degradation of mRNA by RNase H is considered to be the major mechanism of action for antisense oligonucleotides. This technique was originally used to elucidate the function of a target gene, but may also have therapeutic applications, provided it is designed carefully and properly.
The Charge for Going by Foot: Modifying the Surface of Podocytes
Authors: Pavenstädt Hermann
Published: April 2003

The podocyte is a highly differentiated cell which forms a crucial component of the glomerular filtration barrier. It maintains a large filtration surface through the slit membranes and counteracts the distension of the glomerular basement membrane. The podocyte is covered with an anionic glycocalyx believed to be important in the maintenance of foot process structures, but the mechanisms of the cellular interaction between podocyte charge and its function are not clearly understood. It has been speculated that the charge selectivity of the glomerular barrier is influenced by angiotensin II. In experimental models of glomerular nephropathy neutralization of the polyanionic surface with polycations causes a retraction of podocyte foot processes. The effect of polycations is energy and Ca2+ dependent and results in tyrosine kinase induced phosphorylation of proteins of the foot processes. Charge alterations of the podocyte seem also associated with proteinuria in several human glomerular diseases such as membranous or diabetic nephropathy. The knowledge of the interaction between charge and podocyte function might offer new strategies in the treatment of glomerular diseases.
Glycosyl Phosphatidylinositol Anchor
Authors: Kooyman David L. , Byrne Guerard W. , Logan John S.
Published: April 2003

Recently, we and others demonstrated the unique potential for glycosyl phosphatidylinositol (GPI) anchored proteins to transfer from one cell membrane to another in a process we termed ‘painting’. The GPI-anchored proteins were shown to transfer intact and functional. The full significance of this phenomenon has yet to be fully realized, but implications exist in many areas including disease transmission (prions), cell protection (endothelial cells), and senescence (erythrocytes). It is of interest to note that cells exhibiting limited or no biosynthetic capacity (spermatozoa and erythrocytes) have been implicated thus far in cell-cell transfer of GPI-linked molecules. This observation demonstrates the potential for GPI-linked proteins to be ‘painted’ onto cells which otherwise may be incapable of expressing exogenous proteins. We show in this paper that GPI-linked CD59 and decay-accelerating factor will transfer intact from erythrocytes to endothelial cells in transgenic mice. We also demonstrate that the transfer process occurs under physiological conditions using several experimental models including organ and bone marrow transplantation. We detail the procedure to effect transfer of GPI-linked proteins from one cell type to another in either an in vivo or in vitro system.
Potential Functional Roles of Extracellular ATP in Kidney and Urinary Tract
Authors: Chan C.M. , Unwin R.J. , Burnstock G.
Published: April 2003

P2 receptors are sensitive to adenosine triphosphate (ATP) and uridine triphosphate and can be divided into two major subtypes: P2X and P2Y receptors. They are specific membrane-bound receptors which when activated by extracellular ATP initiate a variety of biological effects. The P2X receptors are ligand-gated channels, whereas the P2Y receptors are coupled to G proteins linked to second-messenger systems. In this review, the potential functional roles of extracellular ATP in the kidney and lower urinary tract are discussed and briefly explored in the context of some specific renal and urinary tract abnormalities.
Regulation of the Renal Na/K/2Cl Cotransporter Gene
Authors: Nonoguchi Hiroshi , Itoh Kazuko , Ikebe Mika , Tomita Kimio
Published: April 2003

The recently cloned bumetanide-sensitive Na/K/2Cl cotransporters (ENCC2-3) and thiazide-sensitive Na/Cl cotransporter (ENCC1) have important roles in sodium chloride absorption and secretion, cell volume regulation, ammonium accumulation in medullary interstitium, and ammonium excretion. Genetic mutations of bumetanide-sensitive Na/K/2Cl cotransporters and thiazide-sensitive Na/Cl cotransporter cause the hereditary hypokalemic alkalosis of some Bartter’s syndrome and Gitelman’s syndrome, respectively. The transporters are sensitive to diuretics and several hormones. Dehydration and acidosis influence the gene expression of ENCC3 but not of ENCC1-2. In contrast, sodium loading increases ENCC2 protein expression in the outer medulla.
Early Aldosterone Effects
Authors: Verrey François
Published: April 2003

Aldosterone stimulates sodium reabsorption across tight epithelia via corticosteroid receptors. These receptors are ligand-activated transcription factors which regulate a set of genes, leading to changes in the expression of proteins which are thought to mediate the stimulatory action on sodium reabsorption. This functional response starts after a lag period and can be schematically divided into an early phase of activation and a late phase of accumulation of effectors such as Na+ channels and pumps. Recently, the first gene products regulated sufficiently rapidly by aldosterone to possibly account for the early activation have been identified. However, the actual mediators of the physiological response remain to be determined. Besides these transcriptionally mediated effects, aldosterone has been shown to produce rapid, nongenomic actions on intracellular signalling cascades leading to an activation of Na+/H+ exchange. Such effects are mediated by a different type of receptor and have been described both in classical aldosterone target cells and other cells. Their physiological implications are as yet not clear. The aim of this minireview is to describe briefly both the classical early and rapid nongenomic effects of aldosterone and to address the question whether nongenomic effects might play a role for the Na+ transport response.
Glial Cell Line-Derived Neurotrophic Factor Stimulates Ureteric Bud Outgrowth and Enhances Survival of Ureteric Bud Cells in vitro
Authors: Towers P.R. , Woolf A.S. , Hardman P.
Published: April 2003

Development of epithelial organs requires co-ordinated interactions between epithelial and mesenchymal tissues. Studies using null mutant mice have indicated that the ret receptor and its ligand, glial cell line-derived neurotrophic factor (GDNF), are crucial for initiation of development of the metanephric kidney. However, the role of this signalling system in other branching organs has not been analysed. Here we describe detailed expression studies of ret, GDNF, and a co-receptor for GDNF (GDNFRα) in the developing mouse metanephros, lung, and submandibular salivary gland. Also, we examined the role of this signalling system in the development of these organs in vitro. In situ hybridisation revealed differences in the spatial distribution of the three transcripts in the different organs. At the initiation of metanephric development, late on embryonic day 10 (E10), ret and GDNFRα were detected in the Wolffian duct (including the presumptive ureteric bud) whilst the presumptive metanephric mesenchyme expressed GDNFRα and GDNF. Later in development, all three transcripts were restricted to the nephrogenic zone. In contrast, expression in the lung was not detectable by in situ hybridisation until after initiation of development, at E13.5. At this time ret was expressed throughout the epithelium; GDNF was detected throughout the mesenchyme, and GDNFRα was present in the proximal epithelium and mesenchyme only. Ret and GDNF were not detected in the epithelium or mesenchyme of the developing salivary gland, however, GDNFRα was expressed in the mesenchyme at E13.5 and E16.5. Functional studies demonstrated that in organ culture, GDNF significantly increased branching morphogenesis of the E11.5 metanephros, and induced the formation of ectopic ureteric buds from the base of the bud and from the Wolffian duct. The development of lung and salivary primordia were not affected under similar growth conditions. In a novel ureteric bud primary culture system, GDNF significantly increased cell numbers at 24 and 48 h. In cells cultured on laminin this increase was due to increased survival and proliferation, whereas in cells cultured on fibronectin, only survival was enhanced. Our data suggest that GDNF stimulates outgrowth of the ureteric bud, in part, by enhancing cell survival and possibly by increasing proliferation.
Localization by Immunoelectron Microscopy of Schistosoma mansoni Antigens in the Glomerulus of the Hamster (Mesocricetus auratus) Kidney
Authors: De Brito T. , Carneiro Célia Regina W. , Nakhle Maria Cristina , Lima Dirce M.C. , Abrantes-Lemos Clarice P. , Sandoval Marina , Silva Ana Maria G.
Published: April 2003

Adult worm antigen (AWA) and soluble egg antigen (SEA) were localized ultrastructurally by immunoelectron microscopy using two monoclonal antibodies in the glomeruli of hamsters infected with Schistosoma mansoni cercariae or injected with S. mansoni eggs. AWA was detected in all cercaria-infected groups from the 30th day on and was present mainly in cytoplasm of mesangial cells, mesangial matrix, and glomerular basement membrane, either as isolated gold particles or in small electron-dense deposits of probable immune origin. AWA was encountered also on the inner side of the glomerular basal membrane, close to endothelial cells, and in the foot processes of the glomerular epithelial cells. SEA was detected at similar sites, apparently in lesser amounts, in uninfected hamsters inoculated with S. mansoni eggs into the jugular vein. Schistosomal antigens are apparently processed mainly by mesangial cells which are considered to be critical in the pathogenesis of S. mansoni  associated glomerulopathy. Mesangioproliferative glomerulonephritis, immunoglobulin (IgG and IgM), and C3 deposits were observed in hamsters in which AWA and SEA were visualized. During early phases of the infection and in hamsters in which granulomatous pneumonitis was induced by S. mansoni eggs, glomeruli were unchanged or showed a slight mesangial proliferation. Our findings suggests that egg antigens also contribute to the pathogenesis of experimental glomerulopathy in the hamster.
Imaging Approaches to the Correlation of Structure and Function in the Kidney
Authors: Hare Christopher M.B.
Published: April 2003

Aims: This review sets out to cover the areas of radiology and imaging concerned with the structure and function of the kidney which may not be familiar to those outside the field. Methods: A brief historical introduction to renal imaging is followed in more detail by descriptions of the use of computed tomography in 3D reconstruction, angiography, and functional studies. Comparison with magnetic resonance imaging is made and the use of radiological techniques in intervention. Conclusion: Radiology can provide far more information about the kidneys than mere anatomy. Research promises the possibility of reducing the number of tests required to provide structural and functional information.
Conventional and Confocal Epi-Reflection and Fluorescence Microscopy of the Rat Kidney in vivo
Authors: Boyde Alan , Capasso Giovambattista , Unwin Robert J.
Published: April 2003

To visualize superficial and accessible renal tubule cells functioning in situ and to relate what we can ‘see’ to what we know of their function from more invasive in vivo or less direct in vitro studies means applying and adapting recent advances in epifluorescence and confocal microscopy to improve image resolution and to combine this with the use of fluorescent labels to monitor the handling of specific molecules by the proximal and distal renal tubule cells in vivo. Doing this in living tissue is novel, especially in the kidney. Application of confocal microscopy to the imaging of living tissue, as opposed to isolated cells, has not been widely reported. The kidney surface has been imaged before using the confocal microscope and in preliminary studies we have extended this by using a different confocal system with and without fluorescence. While the studies published up to now have been morphological, comparing standard renal (structural) histology of surface glomeruli and renal tubules with the corresponding in vivo confocal images, more dynamic, real-time studies have been limited. Individual red blood cells can be seen flowing around the peritubule capillary network and nucleated white blood cells can also be distinguished. Tubule cells, endothelial cells, the proximal tubule cell brush border and cell mitochondria can be visualized. Filtration and secretion can be observed, and the early and late parts of the proximal tubule distinguished, and the distal tubule recognized. Localization of fluorescently labeled insulin to the luminal brush border and progressive uptake of label and distribution within proximal tubule cells toward the basolateral (blood side) membrane can be demonstrated. The possibility of monitoring hemodynamic changes and tracking the filtration, uptake, secretion and absorption of fluorescently tagged molecules, as well as intracellular fluorescence, e.g. calcium or pH, is an exciting prospect and is ripe for detailed exploration.
Nuclear Magnetic Resonance Spectroscopy: A Non-Invasive Probe of Kidney Metabolism and Function
Authors: Foxall Peta J.D. , Nicholson Jeremy K.
Published: April 2003

Major advances in nuclear magnetic resonance (NMR) spectroscopic methods and technology have led to the increased use of this technique to study kidney metabolism and function. These studies include: (1) the identification of organic osmolytes in the renal medulla and their role as potential markers of medullary development and damage; (2) changes in renal epithelial cell organic solute transport, such as autosomal dominant polycystic kidney disease, and (3) the biochemical heterogeneity of the nephron and identification of markers of site-specific renal damage in experimental animals and man. The present review summarises these data with the aim of demonstrating how NMR can be used as an indirect, and non-invasive probe of homeostatic mechanisms in vivo and in vitro.
Has the Elusive ‘Natriuretic Factor’ Been Discovered, and If So, Is It a Hormone?
Authors: Wechter William J. , Murray Jr. E. David
Published: April 2003

This review summarizes the interesting and significant papers reported at the International Conference on Natriuretic and Digitalis-like Factors held at the ASH meeting in San Francisco, June 1–2, 1997. This area of investigation has been rejuvenated as of late with near structural determination of two ouabain-like isolates from human plasma (OLF) and bovine hypothalamus (HIF) which are apparently the same compound and the isolation and structural elucidation of a natriuretic metabolite of γ-tocopherol. Spectroscopic information has also been obtained for two other compounds, an ouabain-like factor from bovine adrenals and HHIF from the hypothalamus. An explanation was offered for how low concentrations of digitalis-like factors can regulate vascular reactivity when the predominant isoform of the sodium pump has a low affinity for these compounds. Various groups are examining possible in vivo synthetic pathways that could lead to the production digitalis-like factors. The natriuretic metabolite of γ-tocopherol, LLU-α, fits deWardener’s postulates for a natriuretic hormone and is being examined for its involvement in ECF control. Once the structures for some of these ouabain-like compounds are determined and they are synthesized, these compounds will also be able to be studied employing classical pharmacologic methods.
Defective Proximal Tubule Lysosomal Acidification by Bence Jones Proteins
Authors: Nicastri Ana Lucia , Brandão de Almeida Prado Maria José , Sesso Antonio , Brandão de Almeida Prado Euthymia
Published: April 2003

Proximal tubule handling of two human Bence Jones proteins (neutral and acidic BJP) was evaluated using protein A-gold labelling. After 30 min of acute light-chain infusion into 6 rats (alone or in combination with dinitrophenyl-aminopropyl-methylamine [DAMP]), kidney biopsies were processed for immunoelectron microscopy. Antibodies directed at monoclonal lambda light chains, mannose-6-phosphate cation-independent receptor (MPR) and DAMP were used.Labelling density (number of pA-gold particles/µm2), expressed as median (25–75 percentiles), differed (p < 0.05) between the two BJP, being 94.5 (32.9–212.5) vs. 19.4 (3.7–45.6) pA-gold/µm2 in endocytic vacuoles, and 297.3 (207.1–382.1) vs. 83.2 (16.6–197.0) pA-gold/µm2 in non-vacuolar electrondense endosome-lysosome structures. Labelling density for MPR was 47.7 (22.2–84.6) vs. 4.0 (2.7–6.3) pA-gold/µm2. The area of MPR-labelled structures was also different, i.e.: 0.2 (0.1–0.4) vs. 0.9 (0.5–1.8) µm2. The endosome-lysosome pH distribution range differed significantly: 6.8 (6.4–7.0) vs. 6.3 (5.8–7.0). There was a significant accumulation of neutral BJP in endocytic structures, an acidification deficit of pre-lysosomes/lysosomes and MPR retention, suggestive of defective receptor recycling with this BJP. Interference with the physiological process of lysosomal acidification may be an important mechanism of higher nephrotoxicity in some BJP.
Characterization of B2-Bradykinin Receptors in Rabbit Principal Cells of the Collecting Duct
Authors: Ardaillou Nicole , Placier Sandrine , Zhao Jun , Baudouin Béatrice , Ardaillou Raymond
Published: April 2003

Bradykinin (BK) plays a key role in collecting duct functions. Using an established line of principal cells of the rabbit collecting duct (R.C. SV3), we examined the characteristics of the BK receptors in these cells. [3H]-BK bound specifically to R.C. SV3. Saturation binding analyses allowed KD (968 ± 232 pM) and Bmax values (356 ± 43 fmol/mg protein) to be calculated. Competitive displacement of [3H]-BK was observed with Hoe-140, a specific type 2 BK receptor (BKR-2) antagonist, but not with des arg9-BK, a BKR-1 agonist. The presence of BKR-2 was confirmed by the reverse-transcription polymerase chain reaction technique. BK stimulated cytosolic calcium and inositol phosphate formation in a dose-dependent manner (from 1 nM to 1 µM). BK also inhibited the arginine vasopressin dependent increase of cyclic adenosine monophosphate. This effect could not be related to the production of prostaglandin E2. These results demonstrate the presence of high-affinity BKR-2 in the principal cells of the rabbit collecting duct that are linked to phospholipase C activity and are involved in arginine vasopressin related regulatory loops.
A Comparative Kinetic RT/-PCR Strategy for the Quantitation of mRNAs in Microdissected Human Renal Biopsy Specimens
Authors: Del Prete D. , Forino M. , Gambaro G. , D’Angelo A. , Baggio B. , Anglani F.
Published: April 2003

Molecular biology techniques, to be applicable to a diagnostic renal biopsy specimen, should (1) be highly sensitive to be performed on a very small quantity of tissue; (2) be quantitative because they have to analyze genes normally expressed in the tissue and (3) allow the analysis of as large a number of genes as possible. Among different methods, only the reverse-transcriptase polymerase chain reaction (RT/-PCR) might comply with previous requisites, but the few RT/-PCR examples on renal biopsies in the literature do not allow starting RNA quantification and quality control; furthermore they have the drawback of analyzing only few genes. In an ongoing study to assess the expression of a number of genes in glomeruli and in tubulointerstitium of patients with different nephropathies, we developed a comparative RT/-PCR kinetic strategy based on the purification and quantification of total glomerular and tubulointerstitial RNA and on the use of an internal standard, the housekeeping gene G3PDH. We demonstrate that in microdissected diagnostic renal biopsies (1) glomerular and interstitial starting RNA can be quantified; (2) the G3PDH gene may be used both as an internal standard and as an indirect marker of RNA integrity; (3) as low as 28 ng of total RNA is sufficient to obtain PCR products of eight genes, and (4) it is worth to operate on microdissected biopsy specimens because of the different expression of genes in the two renal compartments.
Modifiable Gene Expression in Mice: Kidney-Specific Deletion of a Target Gene via the Cre-loxP System
Authors: Stec David E. , Sigmund Curt D.
Published: April 2003

With the advent of gene-targeting in mouse embryonic stem (ES) cells, the use of knockout mice to study the physiological effects of loss of gene function has become increasingly prevalent. However, there are several drawbacks with conventional gene-targeting approaches which may make phenotyping of the resultant mice difficult, if not, impossible. Conventional gene-targeting results in the loss of function of the targeted gene in all cells and tissues, which can be problematic for genes which are required developmentally, which exhibit a wide tissue-specific expression pattern, or are part of complex paracrine systems. As with mice that lack the angiotensinogen or endothelin-1 gene, loss of gene function may lead to a lethal phenotype which can be manifested during embryonic development, at birth or postnatally. These limitations could potentially be circumvented by using a system in which the loss of gene function is placed under spatial and/or temporal control. We will discuss how the cre-loxP recombinase system can be applied to delete a gene in a tissue- and developmentally regulated fashion.
Homologous Recombination Based Gene Therapy
Authors: Lai Li-Wen , Lien Yeong-Hau H.
Published: April 2003

Background/Aims: Most of the current expression vector based gene therapy protocols fail to achieve clinically significant transgene expression required for treating genetic diseases. Homologous recombination, initially considered to be of limited use for gene therapy because of its low frequency in mammalian cells, has recently emerged as a potential strategy for developing gene therapy. Methods: Six recent studies of homologous recombination in mammalian cells are reviewed. Different approaches have been used in these studies including RNA/DNA chimeric oligonucleotides, small or large homologous DNA fragments, or adeno-associated viral vectors. Results: Most of these studies show a reasonable frequency of homologous recombination which warrants further in vivo testing. Conclusions: Homologous recombination based gene therapy has the potential to develop into a powerful therapeutic modality for genetic diseases. It can offer permanent expression and normal regulation of corrected genes in appropriate cells or organs and probably can be used for treating dominantly inherited diseases such as polycystic kidney disease.
mRNA Expression of Renal Urea Transporters in Normal and Brattleboro Rats: Effect of Dietary Protein Intake
Authors: Hu Ming Chang , Bankir Lise , Trinh-Trang-Tan Marie-Marcelle
Published: April 2003

Differences in dietary protein level induce differences in fractional excretion of urea, in arginine vasopressin (AVP) plasma level, and in urine concentrating activity (in which intervene the renal urea transporters (UT)). The abundance of mRNA for UT-A1 (of the inner medullary collecting duct, IMCD) UT-A2 (of the descending thin limb) and UT-B1 (of descending vasa recta) was determined by Northern analysis of total RNA extracted from medullary subregions of Sprague-Dawley rats fed for 1 week, a low, normal, or high protein diet. The implication of AVP was then examined by studying AVP-deprived (Brattleboro) rats. Our results show that none of these transporters is affected by the level of protein intake, except UT-A1 that is reduced in terminal IMCD by low protein diet in the absence of AVP (Brattleboro rats). These data suggest that (1) the previously reported effect of kidney medulla hypertonicity on UT-A2 and UT-B1 mRNA expression is somehow obliterated by protein intake deficiency or excess, and (2) AVP influences the mRNA abundance of the UT-A1 of the terminal IMCD during protein deficiency.
Osteopontin Function in Pathology: Lessons from Osteopontin-Deficient Mice
Authors: Rittling Susan R. , Denhardt David T.
Published: April 2003

Osteopontin (OPN) is a soluble secreted phosphoprotein that binds with high affinity to several different integrins. While numerous activities have been ascribed to OPN in vitro, and several in vivo functions have been suggested for the molecule, including much attention focused recently on OPN in different pathologies, the actual role that the protein plays in mammalian physiology remains conjectural. Analysis of recently developed strains of mice lacking OPN because of a targeted disruption of its gene promises to provide important information on this issue. Here, we review the data implicating OPN as functioning in a variety of pathologic conditions as well as the initial results generated with the OPN knockout mice, with particular focus on the question of whether OPN has a protective or a damaging effect in different pathologies. These data will be important to the definition of how OPN contributes to mammalian physiology and pathophysiology.
Cystic Diseases of the Kidney: Role of Adhesion Molecules in Normal and Abnormal Tubulogenesis
Authors: Wilson Patricia D. , Burrow Christopher R.
Published: April 2003

This short review summarizes some information concerning what is known about matrix adhesion molecules, focal adhesion proteins, and cell-cell adhesion molecules in normal renal development and cystic diseases of the kidney. The focus is on human nephrogenesis and disease, but utilizes critical information gained from genetically manipulated mouse models. Interestingly, a significant role for the human PKD-1-encoded gene product, polycystin-1, has been found in cell-matrix interactions via integrins during development, and mutations lead to autosomal dominant polycystic kidney disease (ADPKD). Recent studies on human ADPKD have implicated polycystin-1 in the formation of multiprotein complexes containing focal adhesion proteins at the basal cell surface of the normal ureteric bud. Further evidence of a critical role of cell-matrix interactions via focal adhesion complex formation is provided by the development of renal cystic disease in tensin knockout mice.
Leukocyte-Endothelial Cell Interactions – Lessons from Knockout Mice
Authors: Rosenkranz Alexander R. , Mayadas Tanya N.
Published: April 2003

The advent of gene targeting has led to the generation of several mouse strains deficient in select leukocyte adhesion receptors. These strains of mice have been very informative about the roles of cell adhesion molecules in leukocyte-endothelium interaction and have produced some surprises: roles for leukocyte adhesion receptors have been demonstrated in development as well as pathologies like obesity, and evidence for functional synergies between adhesion receptors have been provided. We attempt in this review to first outline the technique of gene targeting and give an overview of leukocyte adhesion receptors and mice deficient in these receptors. Second, we discuss models of experimental glomerulonephritis and what we have learned about leukocyte adhesion receptors in the pathogenesis of glomerulonephritis through studies in knockout mice.
Mesangial Cells and Their Adhesive Properties
Authors: Pröls Felicitas , Hartner Andrea , Schöcklmann Harald O. , Sterzel R. Bernd
Published: April 2003

Glomerular mesangial cells play a central role in maintaining structure and function of the glomerular capillary ultrafiltration apparatus. Under physiological and pathological conditions, mesangial cells regulate amount and composition of the surrounding extracellular matrix. Conversely, components of the embedding matrix affect the mesangial cell phenotype. These interactions are mediated via specific cell surface receptors, the best studied group of which is the β1 integrin family. The β1 integrins play a role in mesangial cell adhesion, migration, survival and proliferation. Expression and abundance of integrins in healthy and diseased glomeruli and their functions and mediation of signals are discussed in this review. Other factors modulating mesangial cell-matrix interactions, such as antiadhesive proteins, cytokines, disintegrins and nitric oxide, are also considered. The available evidence from in vitro and in vivo studies indicates that receptor-mediated interactions between mesangial cells and the normal or abnormal extracellular matrix regulate the mesangial cell phenotype and thus contribute to normal maintenance of the glomerulus and to remodeling and repair of the glomerular capillary tuft in response to injury.
Diapedesis of Leukocytes: Antisense Oligonucleotides for Rescue
Authors: Dragun Duska , Haller Hermann
Published: April 2003

Ischemia-reperfusion injury is an acute inflammatory process during which leukocytes are intimately involved. In this review, we summarize the current data on the leukocyte cell adhesion cascade in ischemia-reperfusion injury, focus upon studies which have demonstrated specific cell adhesion molecule interactions which mediate the leukocyte involvement in ischemia-reperfusion injury, and suggest future avenues of therapeutic interventions. The increased adhesion between activated vascular endothelium and peripheral blood leukocytes is central to the structural and the functional impairment in ischemia-reperfusion injury. Several families of adhesion molecules, namely the selectins, the intercellular adhesion molecules (ICAMs), and the integrins expressed either on the endothelium or on the leukocytes, are involved the cascade of events. Sequential and overlapping cellular interactions between the members of the three gene families of adhesion receptors result in adhesion of the leukocytes to the endothelium and extravasation at the site of ischemia. The functional importance of ICAM-1 and its β2 integrin ligands in ischemia-reperfusion of the kidney has been demonstrated by monoclonal antibody blockade studies, in knockout mice and by treatment with antisense oligodeoxynulceotides (ODN). We have shown that antisense ODN for ICAM-1 protected the kidney against ischemic renal failure. In addition, in transplanted kidneys, ICAM-1 inhibition by antisense ODN ameliorates ischemia-reperfusion injury and prevents delayed graft function. Recent developments in antisense ODN technology make this a promising therapeutic approach, and antisense ODN treatment of donors or donor organs for ICAM-1 may be useful for the prevention of reperfusion injury in human renal transplantation and could influence acute and chronic graft function.