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  • 1
    Electronic Resource
    Electronic Resource
    Expression of glomerular extracellular matrix components in human diabetic nephropathy: decrease of heparan sulphate in the glomerular basement membrane (1994)
    Springer
    Diabetologia 37 (1994), S. 313-320 
    Details
    ISSN: 1432-0428
    Keywords: Diabetic nephropathy ; heparan sulphate ; heparan sulphate proteoglycan ; glomerular basement membrane ; extracellular matrix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Diabetic nephropathy is characterized by albuminuria which proceeds to overt proteinuria. The highly negatively stained HS side chain of heparan sulphate proteoglycan (HSPG) is a major determinant of the charge-dependent permeability of the GBM. We set out to study the presence of HS and HSPG in the GBM of patients with diabetic nephropathy using newly developed monoclonal antibodies, and to compare HSPG expression to the expression of other previously investigated glomerular extracellular matrix compounds. Immunohistochemically, glomerular extracellular matrix components were analysed in 14 renal biopsies of patients with diabetic nephropathy and compared with those of normal control subjects. Monoclonal antibodies used were: JM403 against the HS side chain of GBM HSPG and JM72 against the HSPG-core protein. Also, a polyclonal antiserum (B31) against human GBM-HSPG-core protein was used. Additionally, antibodies were used against collagen types I, III, IV and against α1(IV)NC, α3(IV)NC and fibronectin. Staining was scored for intensity and for staining pattern by four independent observers who had no previous knowledge of the sample origin. No glomerular staining was seen for collagen type I. Collagen type III was present in some diabetic nodules. Anti-collagen type IV showed a decreased GBM staining in patients with diabetic nephropathy (p = 0.04). With anti-α1(IV)NC no changes in GBM staining intensity were observed; with anti-α3(IV)NC brilliant GBM staining was seen in both groups. Increased mesangial staining (p = 0.003) was seen with anti-collagen type IV in biopsies with nodular lesions. No differences were observed for fibronectin although it was abundantly present in the mesangial area of biopsies from patients with diabetic nephropathy. In biopsies with mesangial expansion and in biopsies with diabetic nodules, we observed a decreased GBM (p = 0.001) HS side chain staining (JM403) without changes in HSPG-core protein staining (JM72,B31). The HS staining pattern regularly changed from a linear to a more granular and irregular pattern. In patients with a creatinine clearance of more than 15 ml/min, the intensity of GBM HS staining showed an inverse correlation with the rate of proteinuria (r = -0.85, p = 0.004), suggesting a functional relationship. The decreased HS staining in the GBM may reflect the potentially disrupted charge barrier in diabetic nephropathy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00398060
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  • 2
    Electronic Resource
    Electronic Resource
    Reduction of heparan sulphate-associated anionic sites in the glomerular basement membrane of rats with streptozotocin-induced diabetic nephropathy (1995)
    Springer
    Diabetologia 38 (1995), S. 1169-1175 
    Details
    ISSN: 1432-0428
    Keywords: Key words Albuminuria ; anionic sites ; cuprolinic blue ; glomerular basement membrane ; heparan sulphate ; rat ; streptozotocin.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Heparan sulphate-associated anionic sites in the glomerular basement membrane were studied in rats 8 months after induction of diabetes by streptozotocin and in age- and sex-matched control rats, employing the cationic dye cuprolinic blue. Morphometric analysis at the ultrastructural level was performed using a computerized image processor. The heparan sulphate specificity of the cuprolinic blue staining was demonstrated by glycosaminoglycan-degrading enzymes, showing that pretreatment of the sections with heparitinase abolished all staining, whereas chondroitinase ABC had no effect. The majority of anionic sites (74 % in diabetic and 81 % in control rats) were found within the lamina rara externa of the glomerular basement membrane. A minority of anionic sites were scattered throughout the lamina densa and lamina rara interna, and were significantly smaller than those in the lamina rara externa of the glomerular basement membrane (p 〈 0.001 and p 〈 0.01 for diabetic and control rats, respectively). Diabetic rats progressively developed albuminuria reaching 40.3 (32.2–62.0) mg/24 h after 8 months in contrast to the control animals (0.8 (0.2–0.9) mg/24 h, p 〈 0.002). At the same time, the number of heparan sulphate anionic sites and the total anionic site surface (number of anionic sites × mean anionic site surface) in the lamina rara externa of the glomerular basement membrane was reduced by 19 % (p 〈 0.021) and by 26 % (p 〈 0.02), respectively. Number and total anionic site surface in the remaining part of the glomerular basement membrane (lamina densa and lamina rara interna) were not significantly changed. We conclude that in streptozotocin-diabetic rats with an increased urinary albumin excretion, a reduced heparan sulphate charge barrier/density is found at the lamina rara externa of the glomerular basement membrane. [Diabetologia (1995) 38: 1169–1175]
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422365
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  • 3
    Electronic Resource
    Electronic Resource
    Reduction of heparan sulphate-associated anionic sites in the glomerular basement membrane of rats with streptozotocin-induced diabetic nephropathy (1995)
    Springer
    Diabetologia 38 (1995), S. 1169-1175 
    Details
    ISSN: 1432-0428
    Keywords: Albuminuria ; anionic sites ; cuprolinic blue ; glomerular basement membrane ; heparan sulphate ; rat ; streptozotocin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Heparan sulphate-associated anionic sites in the glomerular basement membrane were studied in rats 8 months after induction of diabetes by streptozotocin and in age- and sex-matched control rats, employing the cationic dye cuprolinic blue. Morphometric analysis at the ultrastructural level was performed using a computerized image processor. The heparan sulphate specificity of the cuprolinic blue staining was demonstrated by glycosaminoglycan-degrading enzymes, showing that pretreatment of the sections with heparitinase abolished all staining, whereas chondroitinase ABC had no effect. The majority of anionic sites (74% in diabetic and 81% in control rats) were found within the lamina rara externa of the glomerular basement membrane. A minority of anionic sites were scattered throughout the lamina densa and lamina rara interna, and were significantly smaller than those in the lamina rara externa of the glomerular basement membrane (p〈0.001 and p〈0.01 for diabetic and control rats, respectively). Diabetic rats progressively developed albuminuria reaching 40.3 (32.2–62.0) mg/24 h after 8 months in contrast to the control animals (0.8 (0.2–0.9) mg/24 h, p〈0.002). At the same time, the number of heparan sulphate anionic sites and the total anionic site surface (number of anionic sites × mean anionic site surface) in the lamina rara externa of the glomerular basement membrane was reduced by 19% (p〈0.021) and by 26% (p〈0.02), respectively. Number and total anionic site surface in the remaining part of the glomerular basement membrane (lamina densa and lamina rara interna) were not significantly changed. We conclude that in streptozotocin-diabetic rats with an increased urinary albumin excretion, a reduced heparan sulphate charge barrier/density is found at the lamina rara externa of the glomerular basement membrane.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422365
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  • 4
    Electronic Resource
    Electronic Resource
    Expression of glomerular extracellular matrix components in human diabetic nephropathy: decrease of heparan sulphate in the glomerular basement membrane RID=""ID="" 〈e5〉Abbreviations: 〈/e5〉HS: Heparan sulphate; GBM: glomerular basement membrane; HSPG: heparan sulphate proteoglycan; NC: noncollagenous globular domain; IDDM, insulin-dependent diabetes mellitus; NIDDM, non-insulin-dependent diabetes mellitus (1994)
    Springer
    Diabetologia 37 (1994), S. 313-320 
    Details
    ISSN: 1432-0428
    Keywords: Key words Diabetic nephropathy, heparan sulphate, heparan sulphate proteoglycan, glomerular basement membrane, extracellular matrix.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Diabetic nephropathy is characterized by albuminuria which proceeds to overt proteinuria. The highly negatively stained HS side chain of heparan sulphate proteoglycan (HSPG) is a major determinant of the charge-dependent permeability of the GBM. We set out to study the presence of HS and HSPG in the GBM of patients with diabetic nephropathy using newly developed monoclonal antibodies, and to compare HSPG expression to the expression of other previously investigated glomerular extracellular matrix compounds. Immunohistochemically, glomerular extracellular matrix components were analysed in 14 renal biopsies of patients with diabetic nephropathy and compared with those of normal control subjects. Monoclonal antibodies used were: JM403 against the HS side chain of GBM HSPG and JM72 against the HSPG-core protein. Also, a polyclonal antiserum (B31) against human GBM-HSPG-core protein was used. Additionally, antibodies were used against collagen types I, III, IV and against α1(IV)NC, α3(IV)NC and fibronectin. Staining was scored for intensity and for staining pattern by four independent observers who had no previous knowledge of the sample origin. No glomerular staining was seen for collagen type I. Collagen type III was present in some diabetic nodules. Anti-collagen type IV showed a decreased GBM staining in patients with diabetic nephropathy (p =0.04). With anti-α1(IV)NC no changes in GBM staining intensity were observed; with anti-α3(IV)NC brilliant GBM staining was seen in both groups. Increased mesangial staining (p =0.003) was seen with anti-collagen type IV in biopsies with nodular lesions. No differences were observed for fibronectin although it was abundantly present in the mesangial area of biopsies from patients with diabetic nephropathy. In biopsies with mesangial expansion and in biopsies with diabetic nodules, we observed a decreased GBM (p =0.001) HS side chain staining (JM403) without changes in HSPG-core protein staining (JM72,B31). The HS staining pattern regularly changed from a linear to a more granular and irregular pattern. In patients with a creatinine clearance of more than 15 ml/min, the intensity of GBM HS staining showed an inverse correlation with the rate of proteinuria (r = –0.85, p =0.004), suggesting a functional relationship. The decreased HS staining in the GBM may reflect the potentially disrupted charge barrier in diabetic nephropathy. [Diabetologia (1994) 37: 313–320]
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00398060
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  • 5
    Electronic Resource
    Electronic Resource
    Selective proteinuria in diabetic nephropathy in the rat is associated with a relative decrease in glomerular basement membrane heparan sulphate (1995)
    Springer
    Diabetologia 38 (1995), S. 161-172 
    Details
    ISSN: 1432-0428
    Keywords: Glomerular basement membrane ; heparan sulphate ; collagen ; glomerular filtration rate ; albuminuria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In the present study we investigated whether glomerular hyperfiltration and albuminuria in streptozotocin-induced diabetic nephropathy in male Wistar-Münich rats are associated with changes in the heparan sulphate content of the glomerular basement membrane. Rats with a diabetes mellitus duration of 8 months, treated with low doses of insulin, showed a significant increase in glomerular filtration rate (p〈0.01) and effective renal plasma flow (p〈0.05), without alterations in filtration fraction or mean arterial blood pressure. Diabetic rats developed progressive albuminuria (at 7 months, diabetic rats (D): 42±13 vs control rats (C): 0.5±0.2 mg/ 24 h, p〈0.002) and a decrease of the selectivity index (clearance IgG/clearance albumin) of the proteinuria (at 7 months, D: 0.20±0.04 vs C: 0.39±0.17, p〈0.05), suggesting loss of glomerular basement membrane charge. Light- and electron microscopy demonstrated a moderate increase of mesangial matrix and thickening of the glomerular basement membrane in the diabetic rats. Immunohistochemically an increase of laminin, collagen III and IV staining was observed in the mesangium and in the glomerular basement membrane, without alterations in glomerular basement membrane staining of heparan sulphate proteoglycan core protein or heparan sulphate. Giomerular basement membrane heparan sulphate content, quantitated in individual glomerular extracts by a new inhibition ELISA using a specific anti-glomerular basement membrane heparan sulphate monoclonal antibody (JM403), was not altered (median (range) D: 314 (152–941) vs C: 262 (244–467) ng heparan sulphate/mg glomerulus). However, the amount of glomerular 4-hydroxyproline, as a measure for collagen content, was significantly increased (D: 1665 (712–2014) vs C: 672 (515–1208) ng/mg glomerulus, p〈0.01). Consequently, a significant decrease of the heparan sulphate/4-hydroxyproline ratio (D: 0.21 (0.14–1.16) vs C: 0.39 (0.30–0.47), p〈0.05) was found. In summary, we demonstrate that in streptozotocin-diabetic rats glomerular hyperfiltration and a progressive, selective proteinuria are associated with a relative decrease of glomerular basement membrane heparan sulphate. Functionally, a diminished heparan sulphate-associated charge density within the glomerular basement membrane might explain the selective proteinuria in the diabetic rats.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00400090
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  • 6
    Electronic Resource
    Electronic Resource
    Selective proteinuria in diabetic nephropathy in the rat is associated with a relative decrease in glomerular basement membrane heparan sulphate (1995)
    Springer
    Diabetologia 38 (1995), S. 161-172 
    Details
    ISSN: 1432-0428
    Keywords: Key words Glomerular basement membrane ; heparan sulphate ; collagen ; glomerular filtration rate ; albuminuria.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In the present study we investigated whether glomerular hyperfiltration and albuminuria in streptozotocin-induced diabetic nephropathy in male Wistar-Münich rats are associated with changes in the heparan sulphate content of the glomerular basement membrane. Rats with a diabetes mellitus duration of 8 months, treated with low doses of insulin, showed a significant increase in glomerular filtration rate (p 〈 0.01) and effective renal plasma flow (p 〈 0.05), without alterations in filtration fraction or mean arterial blood pressure. Diabetic rats developed progressive albuminuria (at 7 months, diabetic rats (D): 42 ± 13 vs control rats (C): 0.5 ± 0.2 mg/24 h, p 〈 0.002) and a decrease of the selectivity index (clearance IgG/clearance albumin) of the proteinuria (at 7 months, D: 0.20 ± 0.04 vs C: 0.39 ± 0.17, p 〈 0.05), suggesting loss of glomerular basement membrane charge. Light- and electron microscopy demonstrated a moderate increase of mesangial matrix and thickening of the glomerular basement membrane in the diabetic rats. Immunohistochemically an increase of laminin, collagen III and IV staining was observed in the mesangium and in the glomerular basement membrane, without alterations in glomerular basement membrane staining of heparan sulphate proteoglycan core protein or heparan sulphate. Glomerular basement membrane heparan sulphate content, quantitated in individual glomerular extracts by a new inhibition ELISA using a specific anti-glomerular basement membrane heparan sulphate monoclonal antibody (JM403), was not altered (median (range) D: 314 (152–941) vs C: 262 (244–467) ng heparan sulphate/mg glomerulus). However, the amount of glomerular 4-hydroxyproline, as a measure for collagen content, was significantly increased (D: 1665 (712–2014) vs C: 672 (515–1208) ng/mg glomerulus, p 〈 0.01). Consequently, a significant decrease of the heparan sulphate/4-hydroxyproline ratio (D: 0.21 (0.14–1.16) vs C: 0.39 (0.30–0.47), p 〈 0.05) was found. In summary, we demonstrate that in streptozotocin-diabetic rats glomerular hyperfiltration and a progressive, selective proteinuria are associated with a relative decrease of glomerular basement membrane heparan sulphate. Functionally, a diminished heparan sulphate-associated charge density within the glomerular basement membrane might explain the selective proteinuria in the diabetic rats. [Diabetologia (1995) 38: 161–172]
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00400090
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  • 7
    Electronic Resource
    Electronic Resource
    Uber eine temperaturbestandige flussige Phase zur Trennung von Polyolen
    Amsterdam : Elsevier
    Journal of Chromatography A 26 (1967), S. 495-501 
    Details
    ISSN: 0021-9673
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://linkinghub.elsevier.com/retrieve/pii/S0021-9673(01)98910-1
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  • 8
    Electronic Resource
    Electronic Resource
    Effects of Cyclosporin A on Autoimmune Disease in MRL/1 and BXSB Mice (1986)
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 24 (1986), S. 0 
    Details
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: MRL/1 and BXSB mice were treated daily with cyclosporin A (CyA) in an oral dose of 25 mg/kg body weight. With this dose, blood levels within the therapeutic range were obtained. In normal mice CyA in this dose significantly prolonged the survival of an H-2 incompatible skin graft, and suppressed delayed-type hypersensitivity (DTH). It had no influence on the magnitude of a primary antibody response. Autoimmune mice were treated from 6 to 22 weeks of age. CyA treatment did not alter significantly the anti-DNA and anti-IgG autoantibody levels in either strain compared with control mice, who received olive oil. There was a slight but significant increase in serum IgG levels in CyA-treated MRL/1 mice. Clinical signs of glomerulonephritis (decreased kidney function and albuminuria), and glomerular proliferation were not altered by CyA treatment in either strain. The amount of mesangial IgG deposits was reduced in CyA-treated MRL/1 mice, and remained unchanged in BXSB mice. The extent of the interstitial and perivascular infiltrates and the frequency and severity of necrotizing arteritis in the kidneys of MRL/1 mice were reduced by CyA treatment. The most prominent effect of CyA was an evident reduction in lymphoproliferation in MRL/1 mice. Mortality was not reduced by CyA treatment in MRL/1 and BXSB mice.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3083.1986.tb02128.x
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  • 9
    Electronic Resource
    Electronic Resource
    Lückenlos schnittrandkontrollierte Chirurgie am Paraffinschnitt von Melanomen im Gesicht (2000)
    Springer
    Der Hautarzt 51 (2000), S. 826-832 
    Details
    ISSN: 1432-1173
    Keywords: Schlüsselwörter Melanom ; Gesicht ; Schnittrandkontrollierte Chirurgie ; Lokalrezidiv ; Keywords Melanoma ; Face ; Micrographic surgery ; Local recurrence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Abstract Background and objective. Malignant melanomas on sun-exposed skin are often poorly circumscribed and thus recur frequently. The aim of our clinical trial was to compare conventional to a modified micrographic surgery of primary melanoma. Patients/Methods. 28 patients with in-situ (n=7) and invasive (n=21) melanoma were treated with conventional surgical excision with wide margins; eight of these patients developed a local recurrence. In comparison, 20 patients with primary in-situ (n=7) and invasive (n=13) melanoma as well as four patients with recurrence after conventional surgery underwent modified micrographic surgery with delayed closure of the wound. Results. Paraffin specimens of the margins revealed in half of the patients remnants of melanoma although excision was extended to non-lesional skin. Within a mean follow-up period of 21.3 months, none of our patients treated by modified surgery developed a recurrence. Conclusions. Local recurrencies of melanoma on sun exposed skin may be avoided by means of a modified micrographic surgery using permanent histologic sections.
    Notes: Zusammenfassung Hintergrund und Fragestellung. Flächige Melanome sind in aktinisch geschädigter Haut zu den Seiten hin schlecht abgrenzbar und rezidivieren daher häufig. Anhand einer vergleichenden Studie stellen wir die Vorteile einer lückenlos schnittrandkontrollierten Chirurgie bei diesen Melanomen vor. Patienten/Methodik. 28 Patienten mit In-situ- (n=7) und invasiven Melanomen (n=21) wurden mit einem Sicherheitsabstand bis zu 3 cm operiert. Im Folgenden traten bei 8 Patienten Lokalrezidive auf. Im Vergleich zu diesem Kollektiv wandten wir bei 20 Patienten mit vergleichbaren In-situ- (n=7) und invasiven (n=13) Melanomen sowie 4 Patienten mit einem Melanomrezidiv ein lückenlos schnittrandkontrolliertes, 2-zeitiges operatives Vorgehen an. Ergebnisse. Die modifizierte Schnittrandaufarbeitung am Paraffinschnitt zeigte in 50% der Patienten trotz Exzision in klinisch gesunder Haut noch Melanomanteile. Bisher entwickelte keiner der Patienten innerhalb eines mittleren Nachbeobachtungszeitraums von 21,3 Monaten ein Lokalrezidiv. Schlussfolgerung. Die lückenlos schnittrandkontrollierte Chirurgie scheint die hohe Rezidivrate von Melanomen im Gesichtsbereich zu senken.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001050051225
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  • 10
    Electronic Resource
    Electronic Resource
    The glycosaminoglycan content of renal basement membranes in the congenital nephrotic syndrome of the Finnish type (1992)
    Springer
    Pediatric nephrology 6 (1992), S. 10-15 
    Details
    ISSN: 1432-198X
    Keywords: Congenital nephrotic syndrome ; Finnish type ; Glomerular basement membrane ; Heparan sulphate ; Heparan sulphate proteoglycan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A decrease in the concentration of heparan sulphate proteoglycan (HSPG) in the glomerular basement membrane (GBM) is supposed to cause the increased GBM permeability in the congenital nephrotic syndrome (CNS). Therefore, we analysed the glycosaminoglycan (GAG) content and composition of the GBM and tubular basement membrane (TBM) from 3 patients with CNS of the Finnish type (FCNS) and 16 control infants. The GAG content, determined by spectrophotometric assay after papain digestion, was not significantly different in FCNS patients compared with controls. In addition, the GAG composition was comparable in the two groups, with heparan sulphate (HS) constituting at least 75% of the total GAG content. The urinary GAG content (expressed as mg GAG/mmol creatinine) was age dependent, but similar in both groups. Indirect immunofluorescence studies on kidney tissue from normal human infants, using monoclonal or polyclonal antibodies against the core protein of human GBM HSPG, showed linear staining of almost all renal basement membranes. A monoclonal antibody directed against the HS chain of HSPG showed strong GBM and a weak TBM staining. Kidney tissue from three patients with FCNS displayed no discernible differences in the distribution or quality of staining with the same antibodies. These biochemical and immunohistochemical results are in contrast to the decrease in anionic sites (by polyethyleneimine staining) and the replacement of GBM HS by chondroitin sulphate, observed by others in CNS of the diffuse mesangial sclerosis type.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00856820
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