Accumulation of very long-chain fatty acids does not affect mitochondrial function in adrenoleukodystrophy protein deficiency

X-linked adrenoleukodystrophy (X-ALD, OMIM 300100) is a severe inherited neurodegenerative disease, associated with the accumulation of very long-chain fatty acids (VLCFA). The recent unexpected observation that the accumulation of VLCFA in tissues of the Abcd1-deficient mouse model for X-ALD is not due to a deficiency in VLCFA degradation, led to the hypothesis that mitochondrial abnormalities might contribute to X-ALD pathology. Here, we report that in spite of substantial accumulation of VLCFA in whole muscle homogenates, normal VLCFA levels were detected in mitochondria obtained by organellar fractionation. Polarographic analyses of the respiratory chain as well as enzymatic assays of isolated muscle mitochondria revealed no differences between X-ALD and control mice. Moreover, analysis by electron microscopy, revealed normal size, structure and localization of mitochondria in muscle of both groups. Similar to the results obtained in skeletal muscle, the mitochondrial enzyme activities in brain homogenates of Abcd1-deficient and wild-type animals also did not differ. Finally, studies on mitochondrial oxidative phosphorylation in permeabilized human skin fibroblasts of X-ALD patients and controls revealed no abnormalities. Thus, we conclude that the accumulation of VLCFA per se does not cause mitochondrial abnormalities and vice versa-mitochondrial abnormalities are not responsible for the accumulation of VLCFA in X-ALD mice.     

SSCP analysis of paraffin wax embedded tissues in a family with an atypical form of Fabry disease

To investigate the distribution of a single base pair mutation within a family with one known case of Fabry disease, DNA from paraffin wax embedded necropsy material was studied using single-strand conformation polymorphism (SSCP) analysis. The proband, who presented with an atypical form of Fabry disease, had a G to A transition in exon 6 of the α-galactosidase A gene. This patient had mainly cardiac symptoms and late onset disease. Further cases of coronary disorders occurred in this family, including the proband's brother who died at 42 years of age of a cardiac disorder. Formalin fixed, paraffin wax embedded material from the brother and two more distant relatives was available for analysis. SSCP analysis showed that the proband's brother also carried the G to A transition. Thus, the atypical form of Fabry disease and unrelated cardiac diseases with similar clinical symptoms occurred within a single family. The variant form is rare but may account for a few of the numerous cases of cardiac disease in men and should be considered when clusters of cases of cardiac disease occur within a single family.     

Identical abnormality of the short arm of chromosome 18 in two Philadelphia-positive chronic myelocytic leukemia patients with erythroblastic transformation,resulting in duplication of BCR-ABL1 fusion

Two patients with Ph-positive chronic myelocytic leukemia in erythroblastic transformation and rearrangement of the short arm of chromosome 18 are reported. Fluorescence in situ hybridization studies showed that the 18p rearrangement resulted from translocation of the main part of chromosome 22 long arm to 18p, including BCR-ABL1 fusion. The 18p abnormality resulted, thus, in loss of 18p and duplication of BCR-ABL1 in both patients. The possible relation to the erythroblastic type of blastic phase is briefly discussed. In addition an apparently intact germline ABL1 gene was duplicated and inserted into chromosome 6 at band p21 in one of these patients.     

Change of surgical treatment in benign thyroid gland diseases-- influence of surgical management on perioperative complications

Since the beginning of 1994 standardized primary radical resection was performed in benign thyroid diseases at the surgical department, Barmherzige Brueder Hospital, Graz. In multinodular euthyroid goiter bilateral lobectomy or near total lobectomy respectively was done. In case of Graves' disease unilateral lobectomy and contralateral near total resection or "en bloc" thyroidectomy was performed. In accordance with literature a significant reduction of recurrent laryngeal nerve paralysis from approximately 9% to 1.2% occurred in our patients if this surgical procedure was performed consequently. Hypocalcemia rate after surgery needing treatment was approximately 1% at the time of demission.     

Assessment of the viability of haemopoietic cells in the livers of mouse fetuses stored at 4 degrees C. Comparison of various methods of testing their usefulness for haemopoietic transplantation

The usefulness of three different tests available in experimental haematology for the assessment of the viability of fetal liver cells obtained from murine fetuses kept in refrigerator at 4 degrees C was compared. The usefulness of these cells as potential transplants for haemopoiesis reconstitution was assessed in the test with trypan blue, in the test based on clonal growth of GM-CFU in agar, and in the test of the splenic colony forming ability of CFU-S. Fetuses kept in refrigerator at 4 degrees C tested 16 hours after circulation arrest contained still about 70% of the initial number of the haemopoietic stem cells (CFU-S). The proportion of committed cells belonging to the granulocyto-monopoiesis line (GM-CFU) decreased at the same time to about 25% of the initial number, and was a sensitive indicator of hypoxia of the studied organ. The test for the viability of the cells based on the use of trypan blue gave results reflecting better the changes in the number of CFU-S than GM-CFU cells.     

Constitutional C-band pattern in patients with adenomatosis of the colon and rectum

The pattern of polymorphism in the C-band-positive constitutive heterochromatin of chromosomes #1, #9, and #16 was studied in fibroblasts from 23 unrelated patients with adenomatosis of the colon and rectum and in peripheral lymphocytes from 78 control persons. The parameters of the heterochromatic regions analyzed were relative size, symmetry-asymmetry within homologous chromosome pairs, and frequency of inversions. The polyposis coli patients had a significantly higher frequency (p less than 0.05) of partial and total heterochromatin inversion on chromosome #9 than the control group (37.0% compared with 21.8%). In the other parameters studied, no significant differences were found between patients and controls.     

Double wavelength measurement of 3,3',5,5'-tetramethylbenzidine (TMB) provides a three-fold enhancement of the ELISA measuring range

A rapid, simple method is described which permits a three-fold enhancement of the working range of ELISA procedures using TMB as a substrate. This consists of measuring absorbance values at a wavelength away from the absorption maximum of TMB and using a predetermined multiplication factor.     

Diagnostische Bewertung des Nachweises von Gliadin-Antikörpern bei Cöliakie

Zusammenfassung Die Methoden der Komplement-bindung und der Präcipitinreaktion in Agar mit Gliadin als Antigen wurden vergleichend auf ihre Anwendbarkeit in der Diagnostik der Cöliakie geprüft.Beim Präcipitinversuch kamen pseudopositive Reaktionen bei 285 Kontrollen nie vor, bei der Komplementbindung zweimal bei 189 Kontrollen.Zwölf sichere Cöliakiefälle ergaben vergleichend geprüft acht bzw. neun positive Reaktionen, wobei die negativen Befunde für die beiden Verfahren nicht das gleiche Serum jeweils betrafen. Wurde die positive Reaktion in nur einer Methode als ausreichend betrachtet, so war das Ergebnis: elf positive Reaktionen auf zwölf Fälle.Während bei klinisch bestätigter Cöliakie die Übereinstimmung der beiden Methoden gut war, war dies bei den bloßen Verdachtsfällen nicht der Fall. Hier gab es bei der Komplementbindung mehr positive Resultate als bei der Präcipitinreaktion.Es wird vorgeschlagen, beide Methoden nebeneinander anzuwenden und sie bei negativem Ausfall zu wiederholen, wobei die gegebenen diätetischen Richtlinien einzuhalten sind. Als Antigen kann ein Glycerinextrakt aus Gluten verwendet werden, wenn gereinigtes Gliadin nicht zur Verfügung steht. Extrakte mit heißem Wasser geben zu viele pseudopositive Reaktionen.

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Summary Complement fixation tests and precipitin reactions in agar with gliadin as antigen were compared in cases of celiac disease.Pseudopositiv reactions did not occur in 285 controll cases when the precipitin test of Ouchterlony was used, but were seen in two cases of 189 controlls with the complement fixation test.If only one of the two reactions was positive and this was considered as a sufficient proof for the formation of gliadin antibodies those were found in 11 of 12 cases of clinically confirmed celiac disease. In those cases the agreement of the two methods was good, but not so in not confirmed cases where complement fixation tests gave more positive results than the precipitin reaction.It is proposed to use both methods for diagnostic help and to repeat the tests if the reactions are negativ.