Evaluation of route-to-route extrapolation factors based on assessment of repeated dose toxicity studies compiled in the database RepDose®.

The majority of repeated dose toxicity studies are available for the oral route. For risk assessment, however, data are needed from the relevant exposure route, i.e. inhalation or dermal. Instead of conducting additional animal studies, route-to-route (R2R) extrapolation may be performed. To explore uncertainties associated with this approach, we derived extrapolation factors (EF) based on no/lowest effect levels (NOELs/LOELs) in the Fraunhofer RepDose® database. For R2R extrapolation oral-to-inhalation 246 study pairs on 110 chemicals were analyzed. Systemic effects triggered the LOELs in the underlying inhalation studies in 49.2%, local effects in 21.9% and both local and systemic effects in 30.9% of the data pairs. For systemic effects in inhalation studies an EF of 2.2 (95% confidence interval: 1.2-3.1) was derived, for local effects, the EF was 4.4 (95% confidence interval: 2.0-8.6), and the EF without distinguishing local or systemic effects (any EF) was 3.2 (95%, confidence interval: 1.7-5.0). Calculation with LOELs instead of NOELs, exposure duration and intrinsic properties of the chemical (toxicity or physicochemical properties) did not influence the EF significantly. For R2R extrapolation oral-to-dermal 46 study pairs on 28 chemicals were analyzed. An overall EF of 0.4 (95%, confidence interval: 0.2-0.9) was obtained. Here, we found a significant difference of EFs for low and high toxic chemicals. Overall, we conclude that reliable systemic NOELs/LOELs can be obtained for inhalation studies via R2R extrapolation from oral studies. Based on the data for any EF we propose to use an EF of 3, which covers also the uncertainty that unexpected local effects may occur in an inhalation study. For the dermal route, our dataset was too small to allow general conclusions, but the results so far do suggest that the current ECHA guidance is conservative when assuming that dermal absorption is as high as oral absorption.

[Early symptoms and findings in optic nerve meningiomas]. / Erstsymptome und Befunde bei Meningeomen des N. opticus.

PURPOSE OF THE STUDY: While evaluating radiotherapy results in patients with primary and secondary optic nerve sheath meningiomas (ONSM) treated between 1993 and 2002, a large amount of data about early signs and symptoms has been collected which might be helpful for establishing an early diagnosis. METHODS: We have reviewed the charts of the patients available at the Centre of Ophthalmology, collecting especially pretreatment data. RESULTS: 112 patients, 4 with bilateral tumours, 83 % female were included. Mean age was 51.7 years. Visual acuity loss with relative afferent pupillary defect was the main symptom and sign in primary ONSM (38 patients). Median interval between first symptoms and diagnosis was 12 months. Optic discs were in approximately one half atrophic, the other half were swollen, and only rarely normal (3 cases only). Retinociliary shunt vessels were seen in 10 cases. Nerve fibre bundle defects were the major visual field finding (including constriction and central scotoma). Visual acuity was better than 0.5 (20 / 40) in 46 % and worse than 0.1 (20 / 200) in 30 %. In secondary ONSM, the interval to diagnosis was with a median of 6 months shorter than in primary ONSM. Approximately half of the optic discs were atrophic, only 6 % were swollen. Even here nerve fibre bundle defects were dominating, only 7 % had vertical hemianopic defect. Visual acuity was better than 0.5 (20 / 40) in 30 % and in worse than 0.1 (20 / 200) 22 %. 45 % had ocular motility disorders. DISCUSSION: Vertical hemianopic defects were surprisingly rare. The high rate of nerve fibre bundle defects and the relatively high number of patients with good visual acuity might explain why this disorder is occasionally mistaken for glaucoma. A typical clinical appearance can be outlined: mainly mid-aged women, slowly progressing visual loss, frequently motility disorders, relative afferent pupillary defect, nerve fibre bundle defects and atrophic or--mainly in primary ONSM--swollen optic disc form the characteristic picture.

Single preoperative oral application of ascorbic acid does not affect postoperative plasma levels of ascorbic acid.

BACKGROUND AND AIMS: A decrease in ascorbic acid (AA) plasma concentration is well known during the postoperative period and postulated to be caused by increased radical scavenging activity in response to surgical trauma. This often affects postoperative patients and is associated with multiple organ failure. Therefore, substitution of AA could potentially decrease the risk of postoperative complications. This study examines the effect of preoperative oral administration of 1,000 mg AA on the postoperative AA plasma concentration. METHODS: 54 patients were randomly split into two groups; patients in group 1 received no AA preoperatively while group 2 received oral AA (1,000 mg). Plasma samples were obtained preoperatively and on the first postoperative day for AA analysis (HPLC). RESULTS: In both groups the AA concentration was normal preoperatively and reduced postoperatively. CONCLUSION: A preoperative substitution of 1,000 mg AA is not sufficient to prevent postoperative lowered plasma concentration.

Genomic structure of karyopherin alpha2 ( KPNA2) within a low-copy repeat on chromosome 17q23-q24 and mutation analysis in patients with Russell-Silver syndrome.

Human karyopherin alpha2 (KPNA2), a member of the karyopherin alpha family, plays a key role in the nuclear import of proteins with a classical nuclear localization signal (NLS). KPNA2, as part of a karyopherin alpha-beta heterodimer, directly binds to the NLS of proteins and functions as an adaptor that binds NLS-containing proteins via karyopherin beta to the nuclear pore complex. The NLS protein-receptor complex is translocated through the pore by an energy-dependent mechanism. Recently, we have identified and mapped the gene for KPNA2 in close proximity to a translocation breakpoint within 17q23-q24 associated with Russell-Silver syndrome (RSS). Therefore, we considered KPNA2 as a positional candidate gene for this heterogeneous disorder. RSS is mainly characterized by pre- and postnatal growth retardation, lateral asymmetry, and other dysmorphic features. Here, we present the genomic organization of the human KPNA2 gene with 11 exons spanning approximately 10 kb on chromosome 17q23-q24. Screening for mutations within all exons and adjacent intronic sequences from 31 unrelated RSS patients revealed three single nucleotide polymorphisms (SNPs) in exons 1, 5, and 7, and five SNPs in introns 1, 4 (2 SNPs), 8, and 9, respectively. No disease-related mutation was identified by comparing the sequence data of the RSS patients with their clinically normal parents and controls.

Phenotypic findings due to trisomy 7p15.3-pter including the TWIST locus.

We report on a three-month-old boy with a 46,XY,der(Y)t(Y;7)(p11.32;p15.3) karyotype and growth deficiency, postnatal microcephaly with large fontanels, wide sagittal and metopic sutures, hypertelorism, choanal stenosis, micrognathia, bilateral cryptorchidism, hypospadias, abnormal fingers and toes, and severe developmental delay. FISH studies showed partial trisomy 7p resulting from a de novo unbalanced translocation. The application of molecular probes from the TWIST gene region (7p15.3-p21.1) and probes from the pseudoautosomal region (PAR) demonstrated that the 7p15.3-pter fragment was translocated onto Yp with the breakpoint within approximately 20 kb from the Yp telomere. We discuss the possible role of the TWIST gene in abnormal skull development and suggest that trisomy 7p cases with delayed closure of fontanels can be a result of TWIST gene dosage effect.

Kabuki syndrome-like features associated with a small ring chromosome X and XIST gene expression.

Although clinical features in Kabuki syndrome (KS; Niikawa-Kuroki syndrome) have been well defined, the underlying genetic mechanism still remains unclear. We report a 9-year-old girl with typical KS-like facial appearance, skeletal and dermatoglyphic abnormalities, severe mental retardation, and growth deficiency. In 60 of 100 GTG-banded metaphases from peripheral blood lymphocytes, a ring chromosome smaller than a G group chromosome was found, which, according to reverse painting, consisted of Xq11.1q13. The proband's karyotype was described as mos45,X/46,X,+r(X). Several loci were analyzed with fluorescence in situ hybridization (FISH) and microsatellite markers revealing that one r(X) breakpoint mapped proximal to DXS422 (Xp11.21) and the second mapped distal to XIST gene, between loci DXS128E and DXS441 (Xq13.2). Uniparental disomy for X and r(X) was excluded and the paternal origin of r(X) was identified. XIST expression was demonstrated by nested reverse transcription polymerase chain reaction (RT-PCR) using primers spanning exons 5, 6i, and 6 in RNA prepared from lymphocytes. The observation of XIST expression is in contrast to two other cases in which the XIST gene was either not present on r(X) or not expressed. To our knowledge, this is the first case of Kabuki-like syndrome manifestations with r(X) and XIST expression.

Construction of a detailed physical and transcript map of the candidate region for Russell-Silver syndrome on chromosome 17q23-q24.

Russell-Silver syndrome (RSS) is a heterogeneous disorder characterized mainly by pre- and postnatal growth retardation and characteristic dysmorphic features. The genetic cause of this syndrome is unknown. However, two autosomal translocations involving chromosome 17q25 were reported in association with RSS. Molecular analysis of the breakpoint on chromosome 17 of the de novo translocation previously described as t(1;17)(q31;q25) enabled us to refine the localization of the chromosome 17 breakpoint to 17q23-q24. Since no detailed mapping data were available for this region, we established a contig of yeast artificial chromosomes, P1 artificial chromosomes, bacterial artificial chromosomes, and cosmid clones for a 17q segment flanked by the sequence-tagged site (STS) markers D17S1557 and D17S940. This contig covers a physical distance of 4-5 Mb encompassing several novel markers. A transcript map was constructed by assigning genes and expressed sequence tags to the clone contig, and altogether 74 STS markers were mapped. Furthermore, the locus order and content provide insight into several duplication events that have occurred in the chromosomal region 17q23-q24. On the basis of our refined map, we have reduced the translocation breakpoint region to 65 kb between the newly derived markers 58T7 and CF20b. These data provide the molecular tools for the final identification of the RSS gene in 17q23-q24.

Color constancy in goldfish: the limits.

Color constancy was investigated in behavioral training experiments on colors ranging from blue to yellow, located in the color space close to Planck's locus representing the main changes in natural skylight. Two individual goldfish were trained to peck at a test field of medium hue out of a series of 13-15 yellowish and bluish test fields presented simultaneously on a black background. During training the tank in which the fish were swimming freely was illuminated with white light. Correct choices were rewarded with food. During the tests differently saturated yellow or blue illumination was used. The degree of color constancy was inferred from the choice behavior under these illuminations. Perfect color constancy was found up to a certain degree of saturation of the colored light. Beyond this level test fields other than the training test field were chosen, indicating imperfect color constancy. Color constancy was quantified by applying color metrics on the basis of the goldfish cone sensitivity functions.

Simultaneous color contrast in goldfish--a quantitative study.

A set of 9-15 colored test fields was presented to goldfish. In Experiment 1, test field hues ranged from green through yellow to red; in Experiment 2, the hues varied from blue through gray to yellow. In the training conditions, the test fields were presented with a gray or black surround. The fish learned to choose one intermediate test field hue by rewarding them with food. In the test conditions, the color of the surround was changed from gray to green, or red (Experiment 1), and from black to blue, or yellow (Experiment 2). The choice behavior of the goldfish changed substantially: one of the test fields other than the training test field was preferred. Direction and strength of simultaneous color contrast was quantified in goldfish color space. The effect of spatial stimulus configuration was investigated by changing test field size and using narrow annular surrounds. With test field radii ranging between 2 and 7.5 mm simultaneous color contrast was optimal whenever the ratio between surround width and test field radius had a value of about 1:1.

The goldfish--a colour-constant animal.

A series of either thirteen or fifteen coloured test fields with hues from blue through grey to yellow were presented on a black background. Goldfish were trained on a bluish-grey test field by food reward. In the training situation, the setup with the coloured papers was illuminated with white light. In the test situation, the colour of the illumination was changed to blue or yellow. In both test illuminations the goldfish preferred the training field in the same way as under white illumination despite the fact that this test field stimulated the cone types very differently from the training situation. As test fields were present that excited the cones in exactly the same way as under white light, but were not chosen, colour constancy can be concluded. By means of colour metrics, it was possible to quantify direction and strength of colour constancy.

Wavelength discrimination of the goldfish in the ultraviolet spectral range.

Wavelength discrimination ability of the goldfish was measured with a behavioural training technique in the UV spectral range. First, spectral sensitivity was determined for the two fish to adjust the monochromatic lights (between 334 and 450 nm) to equal subjective brightness. The results of the wavelength discrimination experiment show that, independent of which wavelength the fish were trained on, the relative choice frequency reached values above 70% only at wavelengths longer than 410 nm. Wavelength discrimination between 344 and 404 nm was not possible. Accordingly, the delta lambda function increases steeply between 400 and 380 nm, with values between about 12 and 90 nm, respectively. Model computations indicate that the delta lambda function cannot be explained on the basis of the cone sensitivity spectra. Instead, inhibitory interactions have to be assumed which suppress the short wavelength flanks of the short-, mid-, and long-wavelength sensitive cone types in the UV range.

Demonstration of proteases in basal cell carcinomas. A histochemical study using amino acid-4-methoxy-2-naphthylamides as chromogenic substrates.

BACKGROUND: Proteases are reported to play an essential part in the proliferative, invasive, and metastasizing behavior of malignant tumors. The aim of the current study was to determine the activity and localization of proteases in basal cell carcinomas (BCC) histochemically. METHODS: Various proteases were identified histochemically in frozen sections of BCC. The following amino acid-4-methoxy-2-naphthylamides (MNA) were used as chromogenic substrates:alanine-MNA for the detection of aminopeptidase M (APM), glycyl-proline-MNA for dipeptidyl peptidase IV (DPP IV), lysyl-proline-MNA and lysyl-alanine-MNA for dipeptidyl peptidase II (DPP II), glycyl-arginine-MNA for dipeptidyl peptidase I (DPP I), and carbobenzoxy (CBZ)-arginyl-arginine-MNA for cathepsin B. RESULTS: APM activity was high in the peritumorous connective tissue, whereas the tumor epithelium and epidermis had negative results. DPP IV showed a highly positive reaction in both tumor epithelium and surrounding connective tissue. Cathepsin B and DPP I reacted strongly in the tumor epithelium but not in the peritumorous connective tissue. CONCLUSIONS: The marked activity of APM, DPP IV, DPP I, and cathepsin B may be related to the proliferation and invasive growth of BCC. The distribution of the activity of APM and DPP IV indicates dynamic interactions between the tumor epithelium and the adjacent connective tissue in the neoplastic process.

Destruction of tumour parenchyma in basal cell carcinoma by tumour-associated neutral proteases: a histochemical study.

Proteolytic activity was demonstrated histochemically in frozen sections of basal cell carcinomas (BCCs). After incubation of tissue sections in 0.1 M phosphate buffer with 0.25 M NaCl the tumour epithelium was almost completely destroyed. The basal and squamous cell layers of the epidermis disintegrated to varying degrees, particularly where they were directly in contact with tumour epithelium. Serine and metalloprotease inhibitors diminished this tissue destruction. Iodoacetate enhanced tumour destruction, urea and potassium thiocyanate even more so. The high proteolytic activity of BCC demonstrated in this study may be an important factor in the proliferative, invasive and destructive behaviour of this tumour.

[The value of scintimetry compared with histomorphology, parathormone assay, densitometry and radiology in renal osteopathy (author's transl)]. / Die Bedeutung der Szintimetrie im Vergleich mit Histomorphometrie, Parathormon, Densitometrie und Röntgendiagnostik bei renaler Osteopathie.

The distribution of 99mTc-Sn-MDP in the thigh was estimated according to two different indices in 60 normal patients, 21 patients undergoing haemodialysis and one who had had a renal transplant. Deviation of the indices in these patients from the normal are significant. The indices were correlated with respect to their accuracy in detecting renal osteopathy with histomorphometry (as the reference method), radiological diagnosis, densitometry, parathormone assay and other laboratory tests. Accuracy of the method is high and, like histomorphometry, is over 90%.

[Asymptomatic haematuria (author's transl)]. / Die isolierte Hämaturie.

Mesangioproliferative glomerulonephritis of varying severity was found by renal biopsy in 86.2% among 130 patients with asymptomatic haematuria. 3.2% had benign nephrosclerosis, 3.2% had benign nephrosclerosis, 1.6% had interstitial nephritis and one patient showed a previously undiagnosed perimembranous glomerulonephritis. Normal renal parenchyma was observed in only 6.9% of cases. Iummunohistological findings were positive in 25.7% of investigated cases. The majority were IgA deposits combined with IgG and C3 complement. The intensity of haematuria is not correlated with the type and severity of histological changes. "Physiological" haematuria (erythrocytes less than 7/ml) which is usually considered normal must be re-evaluated as a consequence of these histological findings. After exclusion of urological or extrarenal disease only histological investigation of the kidneys will bring the final diagnosis. Repeated radiographical or urological investigations can thus be avoided. The risks of percutaneous renal biopsy under fluoroscopic or sonographic control are decidedly less than the information gained from histological evaluation. The different histological findings indicate that isolated haematuria should be considered only as a symptom and not as a uniform disease indicating focal nephritis.