Urinary sodium dodecyl sulfate electrophoresis with silver staining: a noninvasive diagnostic tool for obstructive uropathy in children.

PURPOSE: Obstructive uropathy such as ureteropelvic junction obstruction in the newborn is a major diagnostic and therapeutic dilemma. We investigated whether urinary sodium dodecyl sulfate electrophoresis with polyacrylamide gel electrophoresis with silver staining could be used to discriminate between children requiring and those not requiring pyeloplasty. MATERIALS AND METHODS: In a pilot study we analyzed the urine of 18 children (mean age 2.7 years) with grade III or IV hydronephrosis according to the Society for Fetal Urology classification. A total of 44 healthy children were studied as controls. Children with hydronephrosis were followed using ultrasound, (99m)technetium mercaptoacetyltriglycine diuretic renography and voiding cystourethrography. Urine was obtained by spontaneous voiding and studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis with silver staining using Melzer's modification. After the study period test results were compared to outcomes, ie whether patients required surgery, and to normalization of previously abnormal protein excretion patterns. RESULTS: All but 1 of the healthy controls had a normal electrophoresis assessment. Of 9 patients followed for hydronephrosis 7 had an abnormal electrophoresis result preoperatively. One child had to be operated on twice because of relapse of ureteropelvic junction obstruction. Six children returned to a normal electrophoresis result postoperatively, including the child who was operated on twice. All children with an initially normal electrophoresis assessment displayed persistent normal values, except 1. Children shifting from a normal to an abnormal electrophoresis result underwent surgery after exclusion of urinary tract infection. CONCLUSIONS: Sodium dodecyl sulfate polyacrylamide gel electrophoresis with silver staining seems to be a good predictive test for clinically relevant ureteropelvic junction obstruction. Further studies are being performed to see whether the test can stand against the gold standard, (99m)technetium mercaptoacetyltriglycine diuretic renography.

Diacap alpha-polysulfone HI PS: a new dialysis membrane with optimum beta2-microglobulin elimination.

BACKGROUND: Plasma concentration of beta2-microglobulin (beta2-m) in the case of renal insufficiency is about 20 to 30 times higher than normal. Beta2-m is associated with secondary amyloidosis, a late complication of regular dialysis therapy. To prevent the complications of secondary amyloidosis beta2-m should therefore be eliminated as efficiently as possible during dialysis treatment. This can be accomplished with dialysis membranes which guarantee sufficient clearance for this molecule. It is a matter of discussion whether removal of beta2-m by dialysis may be able to prevent secondary amyloidosis. METHODS: The dialyzers Diacap HI PS 15 (B. Braun Melsungen) and F70 S (Fresenius Medical Care) were compared in five anuric dialysis patients. Arterial blood was taken at the start and at the end of dialysis. Dialysate samples were taken after 30 and 210 minutes and filtrate samples after 60 and 240 minutes from the start of dialysis. Beta2-m and total protein concentration were measured in plasma, filtrate and dialysate. SDS-PAGE of proteins in the filtrate was carried out and kinetics of beta2-m (Kt/V(beta2-m)) were calculated using the Stiller/Mann model. RESULTS: In both dialyzers beta2-m is detectable at any time in the dialysate leaving the dialyzer. In the filtrate beta2-m concentration is about 10 times higher than in the dialysate. Protein pattern in filtrate of both dialyzers is similar and corresponds to that of the glomerulum filtrate. Beta2-m reduction ratio is slightly lower than urea reduction ratio. Using both dialyzers Kt/V(beta2-m) was 0.80, removing about 60% of the generated beta2-m. CONCLUSIONS: In both dialyzers there is considerable removal of beta2-m. Examination of beta2-m kinetics showed an optimum of Kt/V(beta2) of 0.80 which can not be surpassed. Only 60% of generated beta2-m can be removed by three times per week hemodialysis therapy using high-flux dialyzers.

Pathological excretion patterns of urinary proteins in miners highly exposed to dinitrotoluene.

A cohort of 161 underground miners who had been highly exposed to dinitrotoluene (DNT) in the copper-mining industry of the former German Democratic Republic was reinvestigated for signs of subclinical renal damage. The study included a screening of urinary proteins excreted by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and quantitations of the specific urinary proteins alpha 1-microglobulin and glutathione-S-transferase alpha (GST alpha) as biomarkers for damage of the proximal tubule and glutathione-S-transferase pi (GST pi) for damage of the distal tubule. The exposures were categorized semiquantitatively (low, medium, high, and very high), according to the type and duration of professional contact with DNT. A straight dose-dependence of pathological protein excretion patterns with the semiquantitative ranking of DNT exposure was seen. Most of the previously reported cancer cases of the urinary tract, especially those in the higher exposed groups, were confined to pathological urinary protein excretion patterns. The damage from DNT was directed toward the tubular system. In many cases, the appearance of Tamm-Horsfall protein, a 105-kD protein marker, was noted. Data on the biomarkers alpha 1-microglobulin, GST alpha, and GST pi consistently demonstrated a dose-dependent increase in tubular damage, which confirmed the results of screening by SDS-PAGE and clearly indicated a nephrotoxic effect of DNT under the given conditions of exposure. Within the cluster of cancer patients observed among the DNT-exposed workers, only in exceptional cases were normal biomarker excretions found.

Early Detection of Tubulo-Interstitial Kidney Disease in Children Using Highly Discriminating SDS-Gel Electrophoresis.

Tubulo-interstitial kidney disease is characterized by moderate proteinuria < 1 g/day of low molecular weight proteins in range of MW 10.000-50.000. Even in the physiological proteinuria of < 150 mg/day, tubulo-interstitial kidney disease may exist. Using optimized sodium dodecyl sulfate polyacrylamid gel electrophoresis (SDS-PAGE) according to the method of Melzer, even in proteinuria of less than 150 mg/day all relevant proteins for diagnosis of glomerular or tubulo-interstitial kidney disease can be detected. This study evaluates the tubulo-interstitial kidney disease due to polychemotherapy for different types of cancer in 115 children and in 16 children with pyelo-ureteral junction obstruction. Fifty-two and 63 children were followed up during and after chemotherapy, respectively. During therapy, renal damage was recorded in 43% of patients with leukemia, 56% with nephroblastoma, and 79% with other tumors. Tubular protein patterns were seen up to three years after termination of chemotherapy (25% in acute lymphoplastic leukemia, 35% in nephroblastoma and 62% in other tumors). Patients with persistent complete tubular proteinuria or mixed glomerular/tubular proteinuria were found to have a high risk for irreversible renal failure. Children with congenital pyelo-ureteral junction obstruction could also be classified according to SDS-PAGE protein patterns. Patients without parenchymal lesions did not need surgery. Most of those with pathologic findings in SDS-PAGE exhibited partial or complete remission after surgery. The highly discriminating SDS-PAGE permits a rapid, sensitive, reproducible, and reliable analysis of urine proteins for diagnosis and follow-up of all kinds of congenital or acquired renal parenchymal kidney diseases.

Urine protein analysis by gel electrophoresis and laser densitometry after chemotherapy in pediatric cancer patients.

A common side effect of chemotherapy is reversible or nonreversible nephrotoxicity. SDS polyacrylamide gel electrophoresis combined with laser densitometry was evaluated as a suitable method to analyze pathologic urine proteins. A total of 52 pediatric patients were followed during and 63 patients were followed after therapy. During therapy renal damage was recorded in 43% of the leukemia patients, in 56% of nephroblastoma patients, and 75% of patients with other tumors. Three or more months after therapy pathologic patterns were seen in 25% of acute lymphoblastic leukemia patients, in 35% of patients with nephroblastoma, and in 62% of other patients. Patients with persistent complete tubular proteinuria and mixed glomerular/tubular proteinuria were found to have a high risk for irreversible renal damage and should be controlled periodically. This method permits a rapid and reliable analysis of urine proteins and is suitable for follow-up tests of renal function during and after chemotherapy.

Pathological excretion patterns of urinary proteins in renal cell cancer patients exposed to trichloroethylene.

A study was carried out to investigate urinary protein excretion patterns by means of SDS-polyacrylamide-gel-electrophoresis (SDS-PAGE) in renal cell cancer patients who had previously been exposed to high levels of trichloroethylene. Thirty-eight out of 41 (93%) renal cell cancer patients investigated had former extensive trichloroethylene exposure, but only 23 out of 50 (46%) renal cell cancer patients without a history of occupational exposure to trichloroethylene revealed urinary protein patterns indicative of toxic effects on the tubular system. One hundred controls without histories of overt renal disease and not occupationally exposed to trichloroethylene were examined in the same way; only 11 (11%) of them displayed protein excretion patterns indicative of damage to the renal tubule. These results are supported by alpha 1-microglobulin excretion data. The following conclusions are drawn: (1) Substantially more cases of tubular damage are found amongst renal cell carcinoma patients having been exposed to substantial levels of trichloroethylene over many years as compared with renal cell carcinoma patients not exposed to trichloroethylene. (2) The results support the view that chronic tubular damage is a precondition for the nephrocarcinogenic effect of trichloroethylene. (3) The findings indicate that urine protein patterns, on the basis of the SDS-PAGE methodology, represent a 'biological effect parameter' for the medical surveillance of persons occupationally exposed to trichloroethylene.

Removal of beta 2-microglobulin using grafted affinity adsorbents as therapeutic approach for the treatment of hemodialysis patients.

Patients undergoing long-term hemodialysis are increasingly faced with the problem of developing amyloidosis. This is particularly manifested in a deposition of amyloid in diverse tissues and joints. For the treatment and prevention of amyloidosis, novel affinity adsorbents, which were prepared on the basis of radiation grafted polyamide, have been developed to remove beta 2-microglobulin (beta 2-MG) which is the main constituent of the amyloid. Various affinity ligands, such as alkyl residues with different chain lengths, as well as collagen and gelatin were tested for removing beta 2-MG from human serum and hemofiltrates (HF). Collagen and gelatin carriers show the best adsorption performances. They remove > 95% and > 50% of the beta 2-MG from HF and serum, respectively. The results show that the present approach can be used as the basis for future development of beta 2-MG adsorbents as an additional means in the treatment of dialysis patients.

[Clinical investigations on influence of radiation of venous blood with ultraviolet light on wound healing and postoperative complaints]. / Klinische Untersuchungen zum Einfluss einer Ultraviolett-Bestrahlung von Venenblut (UVB) auf Wundheilung und postoperative Beschwerden.

In 28 patients we performed lower third molar surgery with one following exposition of 50 ml venous blood to ultraviolet light. In 25 other accidentally selected patients we made the same operative procedure with only feigned expodition to ultraviolet rays. After real uv-treatment patients reported significantly less postoperative trouble (pain, swelling, analgetics) than in the control group. Therefore we discuss complex effects of ultraviolet light on wound healing.

[Incidence of abnormalities following the use of hormonal contraceptives]. / Fehlbildungshäufigkeit nach Anwendung hormonaler Kontrazeptiva.

In order to get evidence about the incidence of malformations as because of intrauterine exposition of steroids as after cessation of hormonal contraception in dependence on the time interval between the last intake of hormonale contraceptives (h.c.) and on different pills 5,866 pregnant women have been entered into a case-control-study. 2,834 pregnant women had taken h.c. during pregnancy or got pregnant within 6 menstrual cycles after discontinuation of pills (= cases). For control the data of 3,032 pregnant women were examined. These one had never taken pills or their pregnancy occurred after 7 menstrual cycles after cessation of pill ingestion. Perinatal mortality, Apgar-score (5 min) and incidence of twins were not different in cases and controls. The incidence of malformations was significant (p less than or equal to 0.01) higher in controls than in cases. It depended on age in form of an increase with rising age. The incidence of malformations in both groups increased significantly (p less than or equal to 0.01) with a rising cigarette consumption during pregnancy. The results of this study suggest that smoking regarding malformations represents a higher risk than the intake of h.c.

[Therapeutic results in hyperprolactinemic sterility patients]. / Therapieergebnisse bei hyperprolaktinämischen Sterilitätspatientinnen.

Out of 30 female patients wanting a baby we diagnosed in 27 cases a functional hyperprolactimaemia and in 3 women a macroprolactinom. Therapy was based on bromocryptin, in three cases preceded by a transsphenoidal adenomectomy. We obtained altogether 26 gravidities in 22 patients, including two abortions. 23 children are in good health, one child we lost by haemorrhage as a result of insertion velamentosa.

[Results of endocrinologic diagnosis in sterility patients. A clinical report of experiences]. / Ergebnisse der endokrinologischen Diagnostik bei Sterilitätspatientinnen. Ein klinischer Erfahrungsbericht.

Prolactin and FSH were radioimmunologically found in 16 cases of primary amenorrhoea, 73 cases of secondary amenorrhoea, 27 cases of oligomenorrhoea as well as 22 cases of eumenorrhoea and we diagnosed 36 hyperprolactinaemias, 4 tumours of the hypophysis and 4 hypergonadotropic amenorrheas. In 25 cases of post-pill-amenorrhoeas we found 12 functional hyperprolatinaemias. The prolactinomas were subjected to microsurgical therapy. Functional hyperprolactinaemia was treated with bromocryptin, especially in those cases where a child was wanted.

[Management of functional sterility using clomiphene and cyclophenyl at the Karl-Marx-Stadt District Hospital Gynecologic Clinic]. / Beitrag zur Behandlung der funktionellen Sterilität mit Clomiphen und Cyclophenyl an der Frauenklinik des Bezirkskrankenhauses Karl-Marx-Stadt.

We report on 39 barren patients suffering from anovulatory cycle disturbances or corpus luteum inadequacies. Them were given treatment in the Gynecological Hospital of Karl-Marx-Stadt County at Karl-Marx-Stadt in the past few years and were administered Clomiphendihydrogencitrate or Cyclophenyl to provoke ovulation. Before and during therapy, the total of estrogenes and pregnanediol were determined in the 24h-urine every two days. The clinical data and the results of the hormone investigations were analysed, and an attempt was made to derive a forecast from the estrogene and pregnanediol secretion so as to be able to form an opinion on the positive or negative result of the treatment.

[Precanceroses of the cervix uteri--contribution to diagnosis within the scope of pregnancy interruption]. / Präkanzerose der Cervix uteri - Beitrag zur Diagnostik im Rahmen der Schwangerschaftsunterbrechung

The report deals with the early diagnosis of the carcinoma of the cervix under the aspect of the induced abortion between 1972 and 1974. In 5313 cases of induced abortion, we diagnosed 13 carcinoma of the cervix of the group 0 (0,26%). The medical treatment was carried out by means of conization and hysterectomy, respectively. The significance of the searching for the carcinoma during the pregnancy is mentioned.

PIP: Clinical experience with the diagnosis of very early carcinoma (group 0) of the cervix in 5313 women seeking induced abortion (1972-1974) is reported. Early carcinoma was diagnosed in 13 women on the basis of Papanicolaou smears and colposcopy. (Other epithelial changes were observed in 5 patients.) Abortions were performed in 11 cases, followed by conization (5 cases) or hysteretomy (8 cases). Abortion clinics offer wide opportunities for the diagnosis of early cervical cancer, expecially in younger women.