Kidney-specific cadherin correlates with the ontogenetic origin of renal cell carcinoma subtypes: an indicator of a malignant potential?

INTRODUCTION AND OBJECTIVES: To evaluate retrospectively kidney-specific cadherin (Ksp-cad) expression in renal cell carcinoma (RCC) subtypes and oncocytoma in correlation with its ontogenetic origin of distal and proximal tubules and to correlate Ksp-cad expression with tumour characteristics. MATERIALS AND METHODS: Membranous and cytoplasmic expression of Ksp-cad was determined in 40 clear cell (ccRCC), 25 papillary (pRCC), 19 chromophobe carcinomas (chRCC), 27 oncocytomas (oncocytomas) (n = 111) and 32 benign kidney parenchyma specimens separated in distal tubules (DT) and proximal tubules (PT) by immunohistochemistry using tissue microarray technique. Staining intensity was quantified as a score ranging from 0 to 12. Comparison of data and correlation with tumour characteristics were done by Wilcoxon/Kruskal-Wallis tests (post hoc Tukey-Kramer analysis). RESULTS: In benign renal tissue, membranous and cytoplasmic expression of Ksp-cad in the DT was significantly higher than that in the PT (12.0 ± 0 vs. 5.2 ± 0.3 and 6.3 ± 0.5 vs. 0.0 ± 0.0, respectively; (P < 0.05)). Membranous KSP-cad expression was significantly higher in chRCC (5.2 ± 0.8) and oncocytomas (3.7 ± 0.4) than that in ccRCC (0.8 ± 0.2) and pRCC (1.4 ± 0.4; P < 0.05), while expression between oncocytomas and chRCC did not differ significantly. In RCC, Ksp-cad expression was significantly associated with higher T stage and the occurrence of synchronous metastasis (P < 0.05). Higher N stages and grading tended to correlate with a lower Ksp-cad expression. CONCLUSIONS: In this cohort, the origin of tumour subtypes-chRCC and oncocytomas develop from DT and ccRCC and pRCC from PT cells-is mirrored by the respective Ksp-cad expression. This raises the question whether DT-derived tumours have a less malignant potential than PT-derived tumours.

Metabolism of acrylonitrile by isolated rat hepatocytes.

Several of the pathways of metabolism of the suspected carcinogen acrylonitrile (AN) were identified previously in this laboratory with the use of subcellular fractions and purified enzymes (Guengerich, F.P., Geiger, L.E., Hogy, L.L., and Wright, P.L., Cancer Res., 41: 4925-4933, 1981). In order to establish the relative contributions of the various pathways leading to activated and detoxicated products, we examined AN metabolism in isolated Fischer 344 rat hepatocytes as a model. Reduced glutathione (GSH) was depleted, and cell viability was lost in an AN concentration-dependent manner. The major GSH adduct formed at all AN concentrations was identified as S-(2-cyanoethyl)GSH using thin-layer and high-performance liquid chromatography. Acid hydrolysis and amino acid analysis of labeled hepatocellular, protein revealed S-(2-carboxyethyl)-cysteine as the major adduct formed, indicating direct alkylation of cysteinyl residues by AN. 2-Cyanoethylene oxide accumulated in the hepatocyte incubations but did not appear to contribute extensively to alkylation of GSH or protein. Cyanide, resulting from hydrolysis of 2-cyanoethylene oxide, appeared to be completely converted to thiocyanate, which was identified by gel exclusion chromatography and mass spectrometry of the methyl derivative. The concentration of thiocyanate formed was directly proportional to the concentration of AN used. Cyanide does not appear to play a role in AN-mediated cell death. Alkylation of hepatocellular DNA and RNA and extracellular DNA was not observed to an extent greater than one base in 3.5 X 10(5). The relative rates of the various pathways were compared, and more than 97% of the metabolites can be accounted for by the described reactions. These results are of use in evaluating the contribution of the various pathways and modes of binding of AN to toxicity and carcinogenicity in liver and extrahepatic target tissues.

Specificity of the spermidine requirement for the replication of phi X174 DNA be cell-free extracts of Escherichia coli.

A new experimental approach for assessing the biological significance of spermidine interactions in isolated systems is applied to the stimulation by spermidine of the conversion of phi X174 virion DNA to its replicative form by cell-free extracts of Escherichia coli. At 2.5 mM Mg2+, spermidine activated the reaction 20-fold. Varying the spermidine concentration affected both the rate and extent of this DNA synthetic reaction without altering the nature of the reaction products. We evaluated the biological significance of the spermidine requirement by measuring reaction rates in the presence of a homologous series of spermidine analogs of known activity in vivo. There was a lack of specificity, in that all of these analogs were capable of efficiently substituting for spermidine in stimulating the reaction rate. The relevance of this in vitro spermidine stimulation to Escherichia coli chromosome replication in vivo is discussed in light of the results obtained with the spermidine analogs.

EEG patterns at the time of focal seizure onset.

We studied the electroencephalographic pattern at the time of focal seizure onset in 41 patients. Progressive rhythmic EEG activity was characteristic of spontaneous focal seizures in man. Focal hypersynchrony was present in 49% of cases and served as an accurate localizing indication of cortical irritability. Interictal sharp or spike discharges were absent in 34% of cases. When present, spike and sharp discharges characteristically attentuate at the onset of rhythmic ictal activity. Recognition of the pattern of rhythmic ictal transformation is often essential for the electroencephalographic diagnosis of focal seizures.

Reduced redox state allows prolonged survival of axotomized neonatal retinal ganglion cells.

Axonal injury to CNS neurons results in apoptotic cell death. The processes by which axotomy signals apoptosis are diverse, and may include deprivation of target-derived factors, induction of injury factors, bursts of reactive oxygen species (ROS), and other mechanisms. Our previous studies demonstrated that death of a dissociated retinal ganglion cell, an identified CNS neuron, is ROS-dependent. To better define the mechanisms by which ROS induce retinal ganglion cell death after axotomy, we studied their effects in dissociated neonatal rat retinal cultures. Postnatal day 2-4 Long-Evans rat retinal ganglion cells were retrogradely labeled with the fluorescent tracer 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (DiI). Postnatal day 7-9 retinas were dissociated and cultured in the presence of specific ROS generating systems, scavengers, or redox modulators. Retinal ganglion cells were identified by DiI positivity and viability determined by metabolism of calcein-acetoxymethyl ester. We found that ROS scavengers protected against retinal ganglion cell death after acute dissociation, and the effects of ROS appeared to be due to shifts in the redox potential, as retinal ganglion cell survival was critically dependent on redox state, with greatest survival under mildly reducing conditions. Culture of retinal ganglion cell with the non-thiol-containing reducing agent tris(carboxyethyl)phosphine resulted in long-term survival equivalent to or better than with neurotrophic factors. Our data suggest that axotomy-associated neuronal death induced by acute dissociation may be partly dependent on ROS production, acting to shift the redox state and oxidize one or more key thiols. Understanding the mechanisms by which ROS signal neuronal death could result in strategies for increasing their long-term survival after axonal injury.

Somatostatin receptor scintigraphy in central nervous system tumors: role of blood-brain barrier permeability.

UNLABELLED: Somatostatin receptors are expressed in meningiomas and low-grade gliomas, raising the hope that scintigraphy with 111In-DTPA-D-Phe1-octreotide might be helpful in the in vivo localization, differential diagnosis and postoperative/postradiotherapy brain tumor follow-up. METHODS: Indium-111-DTPA-D-Phe1-octreotide scintigraphy and brain scintigraphy using 99mTc-DTPA as a nonspecific tracer for blood-brain barrier integrity were simultaneously performed in 60 patients with CNS tumors using dual-isotope acquisition mode SPECT. For 23 patients, the scintigraphic findings were also compared with in vitro somatostatin receptor autoradiography of surgical biopsy specimens. RESULTS: In meningiomas (located outside the blood-brain barrier), the somatostatin receptor scan showed all tumors and scintigraphic signal intensity correlating with in vitro SSR density positive in all meningiomas. Less contrast was seen on 99mTc-DTPA scans. In all tumors inside the blood-brain barrier, the 111In-DTPA-D-Phe1-octreotide scan visualized the tumors with a disrupted blood-brain barrier, as seen by 99mTc-DTPA scintigraphy. Discrepancies, however, were observed between somatostatin receptor scintigraphy and in vitro receptor autoradiography. CONCLUSION: Combined somatostatin receptor and 99mTc-DTPA scintigraphy may be helpful for noninvasive differentiation between meningiomas and other CNS tumors. False-negative scans were observed as a result of shielding by the intact blood-brain barrier. Interpretation of negative and positive somatostatin receptor scans in CNS tumors must therefore be done with caution.

Differential susceptibility of retinal ganglion cells to reactive oxygen species.

PURPOSE: Retinal light exposure is a source of oxidative stress, and retinal cells contain molecules that scavenge or inactivate reactive oxygen species (ROS). Yet, ROS also play a role in signal transduction, and some retinal cells (e.g., neurotrophin-dependent retinal ganglion cells, RGCs) may use ROS as part of the signaling process for cell death. RGCs might therefore have specialized mechanisms for regulating ROS levels. The hypothesis that RGCs might regulate ROS differently from other retinal cells was tested by studying their differential response to oxidative stress in vitro. METHODS: RGCs were retrogradely labeled by injecting the fluorescent tracer DiI into the superior colliculi of postnatal day 2 through 4 Long-Evans rats. At postnatal days 7 through 9 the retinas were dissociated with papain and cultured with and without specific ROS-generating systems and/or scavengers. RGCs were identified by their DiI positivity using rhodamine filters. Living cells, determined by metabolism of calcein-AM viewed with fluorescein filters, were counted in triplicate. Degenerate reverse transcription-polymerase chain reaction (RT-PCR) using primers specific to peroxidase homology regions was used to survey for novel peroxidases expressed within normal retinas. RESULTS: Compared with other retinal cells, RGCs were remarkably resistant to cell death induced by superoxide anion, hydrogen peroxide, or hydroxyl radical. Catalase counteracted the effect of each ROS-generating system on retinal cells, consistent with damage occurring via a hydrogen peroxide intermediate. Aminotriazole, L-buthionine sulfoximine, and sodium azide partly abrogated the RGC resistance to oxidative stress, suggesting that this resistance may be mediated by catalase and/or glutathione peroxidase. A limited expression survey within the retina using degenerate RT-PCR did not demonstrate novel peroxidases. CONCLUSIONS: These data suggest a role for one or more endogenous peroxidases within RGCs, which could possibly be protective under conditions of axonal damage. Exploration of the unique characteristics of RGC resistance and susceptibility to injury may help in better understanding the pathophysiology of diseases associated with primary axonal damage.

Renal pathology after arterial yttrium-90 microsphere administration in pigs. A model for superselective radioembolization therapy.

RATIONALE AND OBJECTIVES: To test the selectivity of tissue damage in radioembolization, the authors performed an experimental study using superselective administration of yttrium-90 particles to deliver up to 100 Gy to the porcine kidney. Patterns and severity of damage in test organs were compared with controls, and the feasibility of this model is discussed. METHODS: Eight sows were included in the study. Bio-Rex 70 particles were applied via selective catheterization of the renal artery. Four pigs received inactive particles and four pigs received active particles. Organ distribution and shunting of yttrium-90 were determined, and kidney damage patterns were histologically analyzed. RESULTS AND CONCLUSIONS: The model demonstrates that yttrium-90-labeled resin particles can superselectively be applied. Retention of beta activity in the target organ was more than 95%. In addition to tissue shrinkage from mechanical obstruction, considerable damage ensued mainly by radiation-induced arterial necrosis and arteritis.

An anatomic atlas of the medulla oblongata of the adult goat.

An anatomic atlas of the goat brain stem was developed for use in studies that analyze medullary neuronal groups, and factors that influence variability in the location of neuronal groups were determined. The medullas of 31 adult goats (weight, 17-88 kg) were fixed, harvested, frozen, serially sectioned, stained with 0.5% neutral red, and examined with a light microscope. Obex, the point at which the central canal opens into the fourth ventricle, was taken as the zero reference point from which the rostrocaudal and mediolateral coordinates of medullary neuronal groups were determined, whereas dorsoventral coordinates were calculated from the medullary surface. Histological variations with goat body weight were quantified, and linear regression analysis provided adjustment factors for weight in all three dimensions. Similar analysis of percentage of shrinkage on fixation and processing provided adjustment factors for precise coordinates of medullary neuronal groups. For accurate location of neuronal groups, body weight and histological procedure should be taken into account. The present study provided adjustment factors for body weight and standard histological processing to locate most major medullary neuronal groups in the adult goat.

Multiple rostral medullary nuclei can influence breathing in awake goats.

The purpose of this study was to determine the effect on breathing of neuronal dysfunction in the retrotrapezoid (RTN), facial (FN), gigantocellularis reticularis (RGN), or vestibular (VN) nuclei of adult awake goats. Microtubules were chronically implanted to induce neuronal dysfunction by microinjection of an excitatory amino acid (EAA) receptor antagonist or a neurotoxin. The EAA receptor antagonist had minimal effect on eupneic breathing, but 8--10 days after injection of the neurotoxin, 7 of 10 goats hypoventilated (arterial PCO(2) increased 3.2 +/- 0.7 Torr). Overall there were no significant (P > 0.10) effects of the EAA receptor antagonist on CO(2) sensitivity. However, for all nuclei, > or =66% of the antagonist injections altered CO(2) sensitivity by more than the normal 12.7 +/- 1.6% day-to-day variation. These changes were not uniform, inasmuch as the antagonist increased (RTN, n = 2; FN, n = 7; RGN, n = 6; VN, n = 1) or decreased (RTN, n = 2; RGN, n = 3; VN, n = 2) CO(2) sensitivity. Ten days after injection of the neurotoxin into the FN (n = 3) or RGN (n = 5), CO(2) sensitivity was also reduced. Neuronal dysfunction also did not have a uniform effect on the exercise arterial PCO(2) response, and there was no correlation between effects on CO(2) sensitivity and the exercise hyperpnea. We conclude that there is a heterogeneous population of neurons in these rostral medullary nuclei (or adjacent tissue) that can affect breathing in the awake state, possibly through chemoreception or chemoreceptor-related mechanisms.

Positron emission tomography and ultrasonography. A comparative retrospective study assessing the diagnostic validity in lymph node metastases of malignant melanoma.

BACKGROUND AND DESIGN: A retrospective study involving 20 patients with melanoma with clinically suspicious lymph nodes was conducted to compare the diagnostic validity of fludeoxyglucose F 18 positron emission tomography (PET) and real-time ultrasonography in lymph node metastases of malignant melanoma. RESULTS: A total of 83 lymph nodes were assessed with ultrasonography and PET. Imaging results were confirmed by histologic studies or close follow-up ultrasonographic examinations. Positron emission tomography revealed a sensitivity of 74% and a specificity of 93%. Both investigative methods show comparative sensitivity and specificity. CONCLUSIONS: Ultrasonography is much easier to perform, less time-consuming, and less expensive than PET and it is nonhazardous; therefore, it is ideal for follow-up procedures. Since in routine staging procedures, only sites of expected lymph node involvement are examined, there is a risk of metastases being missed in cases of atypical drainage patterns. Fludeoxyglucose F 18 PET can image proliferating tumors in multiple organ systems and lymph node sites in one session, making it suitable for screening in primary staging procedures and for monitoring response to therapy. Since it is based on metabolic changes, there is good differentiation between scar and tumor tissue. Major disadvantages are restricted access to investigation centers, high imaging costs, and limited anatomical location of metastatic lesions. We conclude that PET does not offer significant advantages in the diagnosis of lymph node metastases compared with ultrasonography.

Surfactants: a survey of short-term genotoxicity testing.

The available results for tests on over 200 surfactants in nine short-term genotoxicity assay systems were reviewed. These tests included the Salmonella/microsome mutation assay, bacterial DNA repair tests, mitotic recombination in Saccharomyces cerevisiae, the mouse lymphoma cell-mutation assay, unscheduled DNA synthesis and sister chromatid exchange assays in mammalian cells, mammalian chromosome damage tests in vitro and in vivo, the dominant lethal test in rodents, and mammalian cell-transformation tests. The collected data cover all four major classes of surfactants: anionic, cationic, nonionic and amphoteric. The results of these genotoxicity tests were overwhelmingly negative. Although there were occasional positive results in bacterial or cell-transformation systems, the testing performed to date indicates that surfactants have negligible potential to cause genetic damage. The available data also indicate that none of the assays were incompatible with testing surfactants for genotoxicity.

[Methods for the superselective radioembolization of liver tumors with 90yttrium-resin particles]. / Methodik der superselektiven Radioembolisation von Lebertumoren mit 90Yttrium-Resin-Partikeln.

An improved technique for radioembolization of non-resectable liver tumours is described. 90yttrium-labelled Bio-Rex-70 particles which show high stability and good tissue tolerance are applied superselectively via the tumour-feeding artery using a microcatheter. The procedure includes both diagnostic and therapeutic angiography. Angiography is complemented by angioscintigraphy (99mTc-MAP) and an i.v. 99mTc-colloid scan of the liver. Thus it is possible pretherapeutically to evaluate tumour volume and dose distribution both in and outside of the liver, and in the lung due to shunt. These estimates were re-evaluated posttherapeutically. Radioembolization can thus be quantified and the doses absorbed by the tumour as well as the liver and lung can be calculated.

Superselective radioembolization of hepatocellular carcinoma: 5-year results of a prospective study.

Twenty patients with unresectable hepatocellular carcinoma (HCC) were followed up to 5 years after transarterial radiotherapy with 90Y-resin particles. Diagnostic radioembolizations of 99mTc-macroaggregates facilitated scintigraphic assessment of activity distribution, dose evaluation and final procedural verification. The overall survival rates were 56, 38 and 14% (after 1, 2 and 3 years, resp.). Patients with unifocal HCC and a single feeding artery (n = 7) even presented 83, 67 and 40% (2 alive after 2.75 and 4 years). With multiple arteries (n = 7), the longest survival was 26 months. Patients with multifocal HCC survived up to 33 months after selective radioembolization. Quality of life was improved in all. Survival was positively correlated with absorbed dose but residual/recurrent tumour occurred even after > or = 300 Gy. Post-treatment symptoms were minimal (35 applications), pulmonary shunt rates were correctly predicted and pulmonary complications avoided.

[Repetitive work and psychosomatic complaints]. / Repetitive Arbeit und psychosomatische Beschwerden.

200 workers of the Swiss watch industry were examined in an interdisciplinary study on the effect of repetitive work on the wellbeing of the worker. Women doing repetitive work with little autonomy complained more often about psychosomatic problems than the male workers doing non-repetitive work. This difference is interpreted as a difference of sexe rather than one of the work situation. However, there is a significant difference in the complaint about nervosity between women being paid monthly and women who were paid by piece or by hour with a premium.

Etiologies of Parkinsonism.

A case report is presented of a patient with Parkinsonism. Three contributory etiological factors are considered with the conclusion that craniocerebral trauma may have been the most significant cause of Parkinsonism in this individual.

[Fluoridation measures and caries prevalence in school children in the former DDR]. / Fluoridierungsmassnahmen und Kariesprävalenz bei Schülern in der ehemaligen DDR.

Statistical analysis of 3,576 DMFT counts from three million observations of children aged 6 to 16 in East Germany shows a strong time-lagged relationship with fluoridation activities. More significant results, however, are found due to age and a score of socioeconomic factors. Additionally, caries prevalence decreases with time. Conversely, DMFT is not affected by size or type of dental staff, neither absolutely nor relatively.

Stimulation of deoxyribonucleic acid replication fork movement by spermidine analogs in polyamine-deficient Escherichia coli.

We examined the rate of deoxyribonucleic acid (DNA) replication fork movement in polyamine-deficient cells of Escherichia coli by two independent techniques. DNA autoradiography was used to directly visualize the length of DNA produced during a given time interval, and replication rates were calculated. The amount of DNA synthesized after blocking protein synthesis also allowed calculation of replication rates. We found that the DNA chain elongation rate in polyamine-deficient cells was about half that of putrescine- or spermidine-supplemented cells. We also found that spermidine homologs of increasing chain length, when present at equal intracellular concentrations, exhibited a decreasing ability to support growth and the rate of DNA replication fork movement. The kinetics of recovery of DNA synthesis from the polyamine-deficient state were also investigated. A new rate of DNA synthesis was reached about 20 min after addition of spermidine to polyamine-limited cells. The rise in the rate of DNA synthesis was preceded by a rise in the intracellular concentration of spermidine.

beta-Endorphin immunoreactive material and authentic beta-endorphin in the plasma of males undergoing anaerobic exercise on a rowing ergometer.

Adrenocorticotropic hormone (ACTH), beta-endorphin immunoreactive material (beta-endorphin IRM), and authentic beta-endorphin (1 -31) have been determined in the plasma of 23 volunteers undergoing anaerobic exercise on a rowing ergometer. The volunteers had different histories of training from occasional physical activities up to intensive preparation for international rowing competitions. ACTH and beta-endorphin-IRM were determined using commercially available immunometric assays; for determination of beta-endorphin (1-31) a highly specific two-site fluid phase immunoprecipitation radioimmunoassay was developed, which did not cross-react with any beta-endorphin derivative or any other opioid peptide tested. In agreement with reports from the literature ACTH and beta-endorphin-IRM concentrations in the plasma rose upon anaerobic exercise in all 23 subjects; this increase in the ACTH and beta-endorphin IRM levels was significantly correlated with the increase of lactate levels observed upon anaerobic exercise. Authentic beta-endorphin (1-31) was only found in two plasma samples containing minor concentrations of the peptide. We conclude that the beta-endorphin immunoreactive material released into blood under anaerobic exercise is identical with authentic beta-endorphin (1-31) only to a minor extent and thus should not be called "beta-endorphin". The major part of the material in fact released into the blood upon anaerobic exercise is probably identical with beta-lipotropin and further components so far unknown.