Ultrasensitive radioimmunoassay of prostate-specific antigen.

We describe a modification of the Yang Pros-check radioimmunoassay for prostate-specific antigen (PSA) that increases the analytical sensitivity of the assay approximately threefold (from a working range of 0.3-50 to 0.1-1.2 micrograms/L). It can detect PSA added to zero-concentration diluent (bovine serum albumin solution) at 0.10 microgram/L or added to zero-concentration control female serum at 0.20 microgram/L (P less than 0.05). In 26 patients tested after cystoprostatectomy for bladder cancer (who had normal prostates without cancer on histologic examination), PSA values by this ultrasensitive assay were all less than 0.10 microgram/L. Therefore, we propose this value as the upper limit of the 95% reference interval. In a retrospective study of two patients who developed recurrent prostate cancer, serum PSA values increased above the 0.1 microgram/L detection limit 175 and 581 days before increasing above the 0.3 microgram/L detection limit of the standard Yang assay. This ultrasensitive radioimmunoassay of PSA should prove more useful than current methods for detecting early recurrence of prostate cancer.     

Endovascular thrombolysis in deep cerebral venous thrombosis.

We present two cases of acute thrombosis of the internal cerebral veins, vein of Galen, and straight sinus without sagittal sinus involvement. Both patients had hydrocephalus and severe edema of the basal ganglia and thalami, one with hemorrhagic infarction of the thalamus. Because both patients rapidly deteriorated to a comatose state, endovascular thrombolysis was performed with urokinase infusion of the deep venous structures. Thrombolysis was continued until a patent channel with brisk flow in the venous structures was achieved. Both patients survived with minimal neurologic deficits.     

The effects of chronic oxotremorine treatment on spatial learning and tolerance development in mice

C57BL mice were treated with 0.5 mg/kg/hr oxotremorine through an implanted subcutaneous cannula for 6 days. Tolerance to oxotremorine was evaluated after treatment by constructing cumulative dose-response curves and measuring body temperature and rotarod performance. At 2 hr after removal, mice exhibited a 15-fold tolerance as measured by body temperature and a 4-fold tolerance as measured by rotarod performance. This tolerance as measured by body temperature was lost by two days after removal from treatment. Immediately after treatment, 3H-QNB binding was reduced in cortex, hippocampus, midbrain, hindbrain, and hypothalamus. Receptors returned to normal within 4 to 8 days after cessation of treatment depending on the brain region. Spatial learning was examined using the Morris water task. Mice that began their training in this task 1 day after they were removed from oxotremorine treatment were impaired in their spatial ability as evidenced by a lack of preference for the trained site during a probe trial. Mice that began their training 2 days after cessation of oxotremorine treatment showed no evidence of impairment in spatial learning. These results suggest that a loss of muscarinic receptors after oxotremorine treatment can be dissociated from tolerance loss and spatial learning deficits.     

Regulation of intestinal guanylate cyclase by the heat-stable enterotoxin of Escherichia coli (STa) and protein kinase C.

The heat-stable enterotoxin of Escherichia coli (STa) stimulates membrane-bound guanylate cyclase in intestinal epithelium and induces fluid and ion secretion. Using the T84 human colon carcinoma cell line as a model, we observed that phorbol esters markedly enhanced STa-stimulated cyclic GMP accumulation in T84 cells (C. S. Weikel, C. L. Spann, C. P. Chambers, J. K. Crane, J. Linden, and E. L. Hewlett, Infect. Immun. 58:1402-1407, 1990). In this study we document that the phorbol ester treatment increases 125I-STa-binding sites as well as membrane-bound guanylate cyclase activity in T84 cells and provide evidence that both effects are mediated by phosphorylation. Guanylate cyclase activity was increased approximately 50% in membranes prepared from intact T84 cells treated with phorbol-12,13-dibutyrate (beta-PDB) and after treatment of homogenates with beta-PDB in a manner dependent on ATP, MgCl2, and cytosol. Similarly, treatment of membranes with purified bovine brain protein kinase C in the presence of appropriate cofactors and beta-PDB resulted in an increase in STa-stimulated guanylate cyclase activity of about 70%. Likewise, the number of 125I-STa-binding sites was increased by about 25 to 40% in membranes prepared from intact cells or homogenates treated with beta-PDB; no effect on binding affinity (Kd = 0.15 nM) was noted. These experiments suggest that protein kinase C may phosphorylate the STa receptor-guanylate cyclase or a closely related protein and increase guanylate cyclase activity. The stimulatory effects of protein kinase C on STa-sensitive guanylate cyclase are opposite in direction to the profound inhibitory effects of the kinase on atrial natriuretic peptide-stimulated guanylate cyclase, demonstrating differential regulation by protein kinases within the guanylate cyclase-receptor family.     

乳腺管状小叶癌

乳腺管状小叶癌（Tubulolobular carcinoma，TLC）最初是被作为小叶癌的管状变型。作者总结了27例TLC的组织学、免疫表型和临床特征，并与纯小管癌和经典型小叶癌进行了比较。此组患者年龄43-79岁（中位年龄60岁）。1例双侧乳腺受累，5例病变为多灶性。肿瘤直径0．5-2．5cm，色灰褐，质硬。组织学观察：TLC的肿瘤细胞形成管状和条索状两种结构模式并相互混杂，且两者比例相当（统称为管状小叶模式）。     

Glomeruläre Veränderungen nach Immunisierung mit heterologem Insulin

Zusammenfassung An 35 Meerschweinchen wurden nach Immunisierung mit reinem Rinderinsulin über 24–150 Tage die glomerulären Nierenveränderungen im Blindversuch an Semidünnschnitten quantitativ untersucht (Mesangiumvolumen, glomeruläre Zellzahl, Index of diabetic glomerular lesions) und, neben der Proteinurie, im Serum die Höhe des Antikörpertiters gegen Insulin (Insulinbindungskapazität) und der Cholesterin-, Harnstoff-und Albuminspiegel bestimmt. Mit zunehmender Dauer der Immunisierung kommt es zu einer Vermehrung der Mesangiumzellen um 57% gegenüber der Norm, zu einer Abnahme der Deckzellen, zu einer Verbreiterung des Mesangiums auf 12,4%, so daß stellenweise noduläre Mesangiumverbreiterungen beobachtet werden. Der IDGL nimmt gegenüber der Norm ab. Antikörper gegen das heterologe Insulin sind in jeder Phase der Immunisierung nachweisbar, während bei den übrigen Serumwerten (Harnstoff, Albumin) nach anfänglichen Veränderungen aller Werte eine spätere Normalisierung eintritt. Der Serumcholesterinspiegel ist auch in den Spätstadien der Immunisierung erhöht. Es kommt somit nach Immunisierung mit heterologem Insulin zu einer Proliferation ortsständiger glomerulärer Zellen im Sinne einer proliferativen intercapillären Glomerulonephritis, die morphologisch an die noduläre diabetische Glomerulosklerose erinnert und deren Ursache in einer Schädigung und funktionellen Störung des Mesangiums zu sehen ist.

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Glomerular changes induced by immunization with heterologous insulin

Summary In a blind-experiment with semi-thin sections we explored quantitatively the glomerular changes (mesangial volume, number of glomerular-cells, index of diabetic glomerular lesions) in 35 guinea pigs after immunizing them with pure beef insulin for 24–150 days. We determined the proteinuria, the serum-levels of the antibody-titer against insulin (insulin-binding capacity) and the levels of cholesterol, urea and albumin. After prolonged immunization the mesangial cells increased 57%. We found a decrease of the epithelial cells, and an increase of the mesangial volume to 12.4%, sporadically of a nodular character. The index of diabetic glomerular lesions decreased. Antibodies against heterologous insulin were demonstrable in every phase of immunization. After initial changes the other serum levels (urea, albumin) returned to normal. The level of serum-cholesterol increased late in immunization.The immunization with heterologous insulin resulted in a proliferation of glomerular cells like in a proliferative intercapillary glomerulonephritis. These changes resembled morphologically the lesions of nodular diabetic glomerulosclerosis and was caused by a functional disorder and injury of the mesangium.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft. Mit technischer Assistenz von Frl. S. Helber und Frl. R. Schnösenberg.

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Nach einem Vortrag auf dem IVth International Congress of Nephrology, Stockholm, Schweden, 23. 6. 69.     

Osmolyte and Na+ transport balances of rat hepatocytes as a function of hypertonic stress

The initial event in the regulatory volume increase (RVI) of rat hepatocytes is an influx of Na+ that is then exchanged for K+ via stimulation of Na+/K+-adenosine triphosphatase (ATPase). In this study, we analysed the activation pattern of the Na+ transporters underlying RVI as a function of the degree of hypertonic stress. In confluent primary cultures, four hypertonic conditions were tested (changes from 300 to 327, 360, 400 or 450 mosmol/l) and the activities of Na+ conductance, Na+/H+ antiport, Na+-K+-2Cl- symport and Na+/K+-ATPase were quantified using intracellular microelectrodes, microfluorometry and time-dependent, furosemide- or ouabain-sensitive 86Rb+ uptake, respectively. Neither Na+ conductance nor Na+-K+-2Cl- symport responded to 327 mosmol/A. At 360, 400 and 450 mosmol/l, uptake via these transporters would lead to increases of cell Na+ by 33.0, 49.0 and 49.0 and by 4.5, 10.4 and 9.2 mmol/l per 10 min, respectively. In contrast, Na+/H+ antiport exhibited 65% of its maximal activation already at 327 mosmol/l. At the four osmolarities tested, this transporter would augment cell Na+ by 6.9, 8.9, 9.8 and 10.6 mmol/l per 10 min. The sums of Na+ import were consistent with the amounts of Na+ exported via Na+/K+-ATPase plus the actual increases of cell Na+ (21.2, 58.5, 63.6 and 68.3 mmol/l per 10 min and 2.2, 4.0, 6.3 and 8.2 mmol/l, respectively). In addition, these elevations of cell Na+ plus the increases of cell K+ (via Na+/K+-ATPase) that amounted to 5.0, 6.5, 17.5 and 18.4 mmol/l were consistent with the increases of intracellular osmotic (cationic) activity of 2.5, 11.5, 21.0 and 28.5 mmol/l, respectively, computed from RVI data. It is concluded that the principle of rat hepatocyte RVI, i.e. an initial uptake of Na+ that is then exchanged for K+ via Na+/K+-ATPase, is realized over the entire range of 9-50% hypertonicity tested. The set-point for the activation of RVI clearly lies below 327 mosmol/l. Na+/H+ antiport is the most sensitive Na+ importer involved in RVI, whereas Na+ conductance plays the prominent role from 360 mosmol/l upwards.