Genetic analysis of Tp53 from urine sediment as a tool for diagnosing recurrence and residual of bladder carcinoma.

OBJECTIVES: Longitudinal study of Tp53 mutation in urine sediments of 26 patients with mutated primary transitional cell carcinoma (TCC) of the urinary bladder at different time periods after transurethral resection of the bladder (TURB), i.e. before and after the first TURB, prior to the control resection and before treatment of a recurrence. METHODS: DNA of the critical Tp53 exons 5-8 was anaylzed by temperature gradients (TGGE) and sequence. RESULTS: (1) In 11 of 12 patients (91.7%) mutation reoccurred with the detection of recurrence of the disease. The mutation frequency in patients without any recurrence was 1 in 8 (12.5%) after a follow-up period of 4-16 months. (2) In 7 of 10 patients, the mutation was no longer detectable in the urine sediment after TURB. (3) The mutation frequency at the control resection 6 weeks after the first TURB was 5 in 7 (71.4%) in patients found to have residual and 1 in 7 (14.2%) in the tumor-free patients. (4) In 9 of 10 samples identical mutations were found by sequence in the recurrent tumor. These results show a significant correlation between the detection of a Tp53 mutation in the urine sediments and tumor recurrence or residual. CONCLUSIONS: (1) Tp53 mutations in the urine sediment could be a useful indicator of tumor recurrence or tumor residual in patients ( approximately 40%) with primary mutated bladder cancer tissue. (2) These results support the monoclonal seeding theory. (3) The finding of identical mutations at different times indicate that the tumor was never totally removed.

p53 tumour suppressor gene mutations in benign prostatic hyperplasia and prostate cancer.

OBJECTIVES: To identify and analyse point mutations in p53 tumour suppressor gene (Tp53) in benign prostatic hyperplasia (BPH) by temperature gradient gel electrophoresis (TGGE) and sequence. MATERIALS AND METHODS: 141 tissue specimens (approx. 100 mg) after transurethral resection of the prostate (TURP), 12 specimens after needle biopsy. Control samples for genetic analysis were (a) 7 prostate tissues without any sign of BPH and malignancy and (b) 103 prostate cancer (PCa) tissues. DNA of the critical Tp53 exons 5-8 was amplified and run on horizontal polyacrylamide gels under defined temperature conditions (TGGE) to yield specific gel shifts and sets of homo- and heteroduplexes in case of mutation. Sequencing with a laser-fluorescent electrophoresis unit was done from re-amplified mutant and wild-type bands. RESULTS: TGGE screening of 153 BPH samples identified 29 specimens with Tp53 mutations (5 in exon 5, 11 in exon 6, 12 in exon 7, 3 in exon 8; 1 tissue sample showed mutations in 3 exons at a time). The computed mutation frequency was 19.0%. Two patients, with mutation in BPH tissue, developed PCa 2-3 years after TURP. One patient with mutation in BPH tissue developed bladder cancer. Of 118 patients with non-mutated DNA in BPH, none is known to have a urological cancer. The Tp53 mutation frequency in 103 PCa samples was 26.2%. Significant differences of mutation frequency between BPH and PCa were detected only in lower exon 5 mutation counts in BPH. CONCLUSION: Tp53 mutation in BPH tissue may be a tumour risk factor.

Pathomorphology of the diabetic kidney.

A short survey is given on important pathomorphologic renal lesions in diabetics, particularly diabetic glomerulosclerosis including aspects on the development of its alterations. The frequency of renal complications in autopsy material of 400 diabetics is reported as well as the occurrence of typical and atypical lesions in a series of 267 biopsy specimens from diabetic patients. In about 60% of this biopsy series semithin sections with silver staining and/or immunohistology could be used which allowed a much more precise diagnosis including the recognition of atypical lesions in diabetics, e.g. glomerulonephritis and the combination of glomerulosclerosis and glomerulonephritis. Therefore, the necessity to apply additional morphological techniques in biopsy material is emphasized.

[p53 in urogenital tumors:analysis of expression and mutation]. / p53 in urogenitalen Tumoren: Eine Expressions- und Mutationsanalyse.

DNA and paraffin material of more than 100 tumors of prostate, bladder and female genital organs were analyzed for p53 aberrations and compared with normal tissues by immunohistochemistry, PCR of p53 exons 5-8 and TGGE. While normal tissues, precancerous and borderline lesions, and well differentiated carcinomas usually showed wild type p53 and negative immunostaining, high grade and/or high stage carcinomas often revealed mutant p53 (rate of mutation in exon 8 >> 7 >> 6 >> 5) and/or p53 accumulation. Accumulation of p53 protein in the absence of detectable mutant p53 was recognized more often in prostate cancer than in any other tumor examined. Although p53 aberration probably represents a late molecular event in cancerogenesis, its detection may be of clinical interest as genetic footprint in recurrent and metastatic disease.

[Analgesic nephropathy (AN) with special reference to capillary sclerosis. A multicenter study from the former East Germany]. / Zur Analgetika-Nephropathie (AN) unter besonderer Berücksichtigung der Kapillarsklerose. Eine Multicenter-Studie aus der ehemaligen DDR.

The incidence of capillary sclerosis in the mucosa of the upper urinary tract was beyond expectation, according to a multicenter study conducted at 11 pathological institutes in all regions of the former GDR from which the following more specific findings were obtained: Capillary sclerosis was recorded primarily from women (1.4:1) in 3.6% of 3,929 autopsy cases (minimum age being 40 years). This result has close to the outcome of a study conducted in Basle, Switzerland, though highest severity (0.33%) and the complete morphological picture of analgesic nephropathy (0.54%) were clearly less frequent, as compared to the above Swiss findings. No reliable conclusion can as yet be drawn regarding the infrequent case of renal pelvis carcinoma. Epidemiological and clinical studies are likely to suggest that an increase in findings might be expected in this part of Germany and might be aetiologically attributable to abuse of analgesics.

[Use of OKT-3 in treatment of rejection following kidney transplantation]. / Einsatz von OKT-3 bei der Rejektionsbehandlung nach Nierentransplantation.

Application of OKT-3 as rescue therapy in steroid-resistant rejection was effective in 7 out of 9 patients early after cadaveric kidney transplantation. Under attention of instructions for use of OKT-3 the adverse first-dose reactions are rare. The problem of OKT-3 therapy is early use and its reuse.

[Results of conversion from conventional immunosuppressive therapy to cyclosporin A in complications following kidney transplantation]. / Ergebnisse der Konversion von konventioneller immunsuppressiver Therapie auf Cyclosporin A bei Komplikationen nach Nierentransplantation.

In 41 patients suffering from complications late after kidney transplantation the conventional immunosuppression was converted to Cyclosporin A. A complete normalization was obtained in the case of acquired immunoglobulin G deficiency and in persistent leukocytopenia. 6 patients suffering from life-threatening bacterial infections did survive after conversion with functioning allograft and 8 out of 9 late rejections were reversible. Conversion may be necessary in chronic hepatopathy and chronic rejection. A conversion to cyclosporin A is recommended because of control of complications in 78% (46/59) of the recipients.

[Immunohistologic differentiation of interstitial infiltration cells in biopsies of transplanted kidneys]. / Immunhistologische Differenzierung interstitieller Infiltratzellen in Biopsien transplantierter Nieren.

Despite known morphological criteria of kidney allograft rejection a definite diagnosis of interstitial rejection in not possible in each case. Therefore, by an enzyme histological differentiation of infiltrating cells in the transplant biopsy specimen this problem should be solved. It was used the PAP method against pan T lymphocytes, T4 and T8 lymphocytes, monocytes and granulocytes. An acute interstitial rejection can be verified if 1. the number of lymphocytes and their subpopulations did increase over the maximum values, 2. without vascular rejection is the T4/T8 ration less than 1 and 3. in the presence of vascular rejection is the T4/T8 ratio greater than 1. Additional information about cyclosporin injury or viral infections may be possible. It is possible increase markedly the rate of safer diagnosis by immunohistological differentiation of infiltrating cells.

[Kidney biopsies in diabetic patients--value of semi-thin layer study]. / Nierenbiopsien von Diabetikern--Wert der Semidünnschnittuntersuchung.

Kidney biopsies of diabetics were divided into 2 groups: 113 biopsies with conventional paraffin sections (group I) were compared with 154 biopsy specimens with additional examination by semithin sections (group II). In group II glomerular lesions were observed in 81.3%, i.e. about twice as frequently as in group I or compared to the diagnoses made from the paraffin sections in group II, respectively. Most often additionally, diffuse glomerulosclerosis or glomerulonephritis were detected, partly also in combination.

[De novo diabetic glomerulosclerosis in kidney transplants]. / Diabetische Glomerulosklerose de novo in Neirentransplantaten.

Report on 3 cases who had been non-diabetics at the time of cadaver kidney transplantation. After that within 1-5 years diabetes mellitus developed. The patients died 13-14 years after transplantation (9-12 years of diabetes duration). Autopsy revealed typical nodular glomerulosclerosis including diffuse mesangial widening, glomerular aneurysms, exudative lesions, capsular drops, arteriolosclerosis, and in 2 cases with hyalinosis of the vas efferens; moreover, chronic transplant glomerulopathy was found in all cases. Steroid therapy seemed to be responsible for the diabetic state, in 2 cases apparently in combination with disposition.

[Glomerulonephritis (GN) in childhood. II. Epidemiology and clinical aspects of various histological forms of glomerulonephritis]. / Zur Glomerulonephritis (GN) im Kindesalter. II. Mitteilung: Epidemiologie und Klinik verschiedener histologischer Glomerulonephritisformen.

From 101 children with histological verified glomerulonephritis the following data were analyzed according to the morphological alterations: age distribution, symptoms and onset of the disease, previous diseases, and observation time. Differences of these parameters were found firstly between minimal glomerular changes and proliferative lesions. Within the various proliferative forms the differences were minimal. In children with a diffuse mesangio-proliferative glomerulonephritis and MPGN the characteristics of blood pressure, proteinuria and hematuria were observed during a follow-up time of 5 years. In conclusion, a distinction between single types of glomerulonephritis could be made exceptionally without examination of kidney function and morphology.

[Glomerulonephritis in sarcoidosis]. / Glomerulonephritis bei Sarkoidose.

The renal involvement in sarcoidosis can be demonstrated as nephrocalcinosis, nephrolithiasis, granulomatous nephritis, and glomerulonephritis. The clinical signs of a renal manifestation are inferior than the morphological findings, so that the reported frequency of 9-25% is an approximate value. In the literature the glomerular changes are rare. In the case of glomerulonephritis the membranous form is dominating; an extracapillar-proliferative glomerulonephritis is described in only 3 cases in the literature. The course of a 16-year-old male patient with an extracapillar-proliferative glomerulonephritis and M. Boeck is described in detail. After a prednisolone therapy for 2 years, which was introduced by methylprednisolone at 1000 mg for 3 days, the renal function was improved continuously.

[Relations between vascular, interstitial and tubular changes in the transplanted kidney]. / Beziehungen zwischen vaskulären und interstitiellen sowie tubulären Veränderungen im Nierentransplantat.

To determine the question of the extent of fibrous intimal swelling in chronic transplant vasculopathy, of fibrous interstitial broadening and of tubular atrophy, we performed semi-quantitative histological examinations on 139 biopsy specimens, 70 removed kidney allografts and 35 allografts after autopsy, respectively. A positive correlation was found between vascular and interstitial changes. Morphometric analyses were done in 9 removed transplants and 2 normal kidneys, at which in each case 4 cortical regions including the supplying middle-sized and small arteries are examined. It was shown, that the extent of intimal swelling was variable and in the same sense also the degree of interstitial proliferation. Thus, the correlation as mentioned above is confirmed. Therefore, it is presumed that the extent of chronic transplant vasculopathy with reduction of the blood supply is most important to the formation of interstitial broadening and tubular atrophy. On principle other causes are possible and must be excluded by histological and/or angiographic investigations.

[Autopsy analysis of the cause of death in patients with kidney transplants]. / Autoptische Analyse det Todesursachen bei Nierentransplantierten.

68 cases of autopsy of 950 patients who underwent a kidney transplantation in Berlin from 1970 to 1986 were analysed with regard to their main disease, the disease directly leading to death and secondary disease. The average age of the deceased was 39.6 years (15-56 years), the duration between kidney transplantation and death was 51.2 months (1-192 months). In the first place there are diseases of the liver (30 times primary disease, 28 times secondary disease), in which cases 24 times a liver cirrhosis was existing and in 11 cases a coma hepaticum caused death. Apart from this septic-septicopyaemic processes (23 times) were of great importance for the exitus, in which cases there were frequently close relations to liver diseases (10 times liver diseases primary disease, 12 times secondary disease). Furthermore are important for the occurrence of death the renal hypertension as well as hemorrhages particularly in the gastrointestinal tract. 15 times a diabetes mellitus was stated, in 13 cases severe changes of bones and joints were diagnosed. 4 times a neoplasia was present (3 times immunocytoma, once liver carcinoma). The diseases diagnosed are to be regarded as a sequela of the primary disease, above all, however, caused by the long-lasting immunosuppressive therapy and the influence on the endocrine system.

[Glomerulonephritis (GN) in childhood. I. Epidemiology, clinical aspects and kidney function in 125 children]. / Zur Glomerulonephritis (GN) im Kindesalter. I. Mitteilung: Epidemiologie, Klinik und Nierenfunktion von 125 Kindern.

The blood pressure, urinary symptoms (proteinuria, hematuria, casts), the albumin- and cholesterol concentration in the serum and the renal function (GFR, RPF) of 125 children with chronic glomerulonephritis (GN) were analysed. 76 children had only single symptoms (93% proteinuria, 33% hypertension), which indicated a GN. The serum albumin and cholesterol concentration were pathological in 43% of the patients and serum creatinine level was pathological in 20% of the children. After 6 years the individual courses of renal function demonstrated a deterioration of GFR and RPF for most of the children. It can be concluded, that the summary consideration of epidemiology, symptomatology and renal function of different glomerular lesions has only a limited application to the clinical practice.

[Results of percutaneous biopsy of kidney transplants. Analysis of 200 successful puncture biopsies]. / Ergebnisse der perkutanen Biopsie von Nierentransplantaten. Analyse von 200 erfolgreichen Punktaten.

During a period of about 4 3/4 years 200 biopsy specimens (= 100%) with judgeable tissue of kidney grafts were examined; from these 99 had been performed during the first 6 months. The best assessment concerned the interstitial cellular rejection which was observed in 73.5%, furthermore, in 9.5% cellular infiltrates of uncertain pathogenesis were found. In 19.5% acute subacute and in 35% chronic transplant vasculopathy occurred while other vascular lesions were relatively rare. Transplantat glomerulopathy was diagnosed in 25% and glomerulonephritis in 14.5%--mostly de novo. In further 26% different minor glomerular abnormalities and in 14.5% chronic ischaemic glomeruli (mostly focally) were observed. The assessment was limited in those cases with lack of arteries (15.5%) or glomeruli (8.5%) within the specimen. However, by judging tubular atrophy and interstitial fibrosis certain conclusions on chronic stenosing arterial processes may be drawn. In the early stage usually the condition of the parenchyma can be assessed well which allows some recommendation for further treatment (conservative therapy or ectomy). On the whole renal biopsy is the best method to assess the state of the transplant.

[Antiglomerular basement membrane antibody-mediated nephritis: incidence and morphological appearance in biopsy materials]. / Anti-Glomerulusbasalmembranantikörper-Nephritis. Häufigkeit und morphologische Erscheinungsformen im Biopsiematerial.

A glomerulonephritis (GN) of the basement membrane antibody-mediated type was diagnosed in 51 patients (44 men, 7 women) on the basis of immunohistological findings in renal biopsy material in a period of 15 years. This represented about 5% of the cases in which immunohistologic studies gave evidence of glomerular disease. Young men, about 20 years old, were especially affected: 35 patients were men between ages 16 and 25 (69%). In about half the cases there were also pulmonary manifestations (Goodpasture's syndrome). In 63% of the cases there was extracapillary GN with compression and/or marked destruction of the glomerular tuft. Anti-glomerular basement membrane antibody-mediated nephritis was the most frequent cause of a rapidly progressive extracapillary GN in our sample. The broad use of renal biopsy in Goodpasture's syndrome has demonstrated that the morphologic spectrum of renal manifestations is much wider than originally believed. It ranges from normal or minimal findings and mild mesangioproliferative or focal extracapillary GN to severe diffuse extracapillary GN. The rapidly progressive form of extracapillary GN was not significantly more frequent in Goodpasture's syndrome than in the cases of GN without lung findings (62%). Immunohistologic findings apparently can persist unchanged for months and therefore microscopic examinations do not allow reliable conclusions concerning the natural history of immunologic activity in this process to be drawn.

[Pathomorphological differential diagnosis of glomerular lesions in diabetics]. / Pathomorphologische Differentialdiagnose der Glomerulus-veränderungen bei Diabetikern.

In renal biopsy specimens taken from 165 diabetics 3 cases with amyloidosis and 25 with glomerulonephritis (GN) were found. Among these were 13 cases with membranous and 10 with mesangial proliferative GN. This remarkably high rate may be partly due to selection, but, it can also be explained by the techniques of investigation because most of the GN cases were diagnosed by semithin sections and/or immune histology, respectively. In paraffin sections these alterations may be misinterpreted as diffuse glomerulosclerosis (GS), borderline lesions or normal findings. This is understandable because in mesangial proliferative GN, as well as in diffuse GS, the mesangium is broadened; similarly the spike formation or basement membrane doubling of membranous GN may simulate a thickening of the basement membrane. In addition, the differential diagnosis between GS and other diseases/lesions is discussed. We conclude that the examination of specimens by adequate methods will reveal "nondiabetic" glomerular lesions in diabetics more frequently than previously reported in the literature.