Biomedical subjects
Publications and source records attributed to M Molnar.
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Our work and that of others defined mitosis-specific (Rad21 subfamily) and meiosis-specific (Rec8 subfamily) proteins involved in sister chromatid cohesion in several eukaryotes, including humans. Mutation of the fission yeast Schizosaccharomyces pombe rec8 gene was previously shown to confer a number of meiotic phenotypes, including strong reduction of recombination frequencies in the central region of chromosome III, absence of linear element polymerization, reduced pairing of homologous chromosomes, reduced sister chromatid cohesion, aberrant chromosome segregation, defects in spore formation, and reduced spore viability. Here we extend the description of recombination reduction to the central regions of chromosomes I and II. We show at the protein level that expression of rec8 is meiosis specific and that Rec8p localizes to approximately 100 foci per prophase nucleus. Rec8p was present in an unphosphorylated form early in meiotic prophase but was phosphorylated prior to meiosis I, as demonstrated by analysis of the mei4 mutant blocked before meiosis I. Evidence for the persistence of Rec8p beyond meiosis I was obtained by analysis of the mutant mes1 blocked before meiosis II. A human gene, which we designate hrec8, showed significant primary sequence similarity to rec8 and was mapped to chromosome 14. High mRNA expression of mouse and human rec8 genes was found only in germ line cells, specifically in testes and, interestingly, in spermatids. hrec8 was also expressed at a low level in the thymus. Sequence similarity and testis-specific expression indicate evolutionarily conserved functions of Rec8p in meiosis. Possible roles of Rec8p in the integration of different meiotic events are discussed.
Intracellular and extracellular recordings were used to assess the cholinergic function in hippocampal slices from juvenile rats chronically deprived of NGF. NGF was neutralised by implanting into the lateral ventricle of postnatal (P) day 2 rats, alphaD11 hybridoma cells (secreting monoclonal antibodies specific for NGF). Parental myeloma cells (P3U) were used as controls. At P15-P18, slow cholinergic EPSPs could be elicited in cells from both alphaD11- and P3U-treated rats. However, slices from alphaD11-implanted rats exhibited a 50% reduction in acetylcholine release following stimulation of cholinergic fibres. This effect was associated to a significant increase in the sensitivity of pyramidal cells to carbachol, as suggested by the shift to the left of the dose/response curve. This may reflect a compensatory mechanism for the reduced efficacy of cholinergic innervation in NGF-deprived rats. In both alphaD11- and P3U-treated rats, carbachol was able to induce a similar concentration-dependent depression of the field EPSPs, evoked by Schaffer collateral stimulation, suggesting that presynaptic muscarinic receptors were not altered. In rats implanted with alphaD11 cells at P15 and sacrificed at P21-P24, no changes in the sensitivity to carbachol were found. At this developmental stage, no differences in acetylcholine release were observed between P3U- and alphaD11-treated animals. These results provide physiological evidence for a regulatory role of NGF in the cholinergic function of the hippocampus during postnatal development.
The aim of the study was to test the clinical value of attenuation-corrected (AC) thallium-201 single-photon emission tomography using a moving gadolinium-153 line source in a group of patients in whom coronary angiography was planned because of clinically suspected coronary artery disease (CAD). Furthermore, we wanted to test the impact of AC on assessment of the extent and severity of perfusion abnormalities. A total of 107 patients planned to undergo coronary angiography were included in the study. In each patient, AC and NC (non-corrected) 201Tl SPET was performed. AC and NC images were evaluated visually as well as by a 31-segment semiquantitative analysis and the findings were correlated with angiographic results. Patients were assigned to two groups: group A with angina and no previous cardiac infarction or intervention and group B with known CAD because of previous myocardial infarction or intervention. With visual analysis, NC revealed a sensitivity of 88.9% in group A and 74.3% in group B, compared to 94.4% in group A and 94.3% in group B with AC. Specificity for NC was calculated to be 68.7% for group A and 91.3% for group B. AC demonstrated significantly higher specificity of 83.9% and 100% respectively. This effect was particularly demonstrated for males and bicycle workload. The extent and severity of perfusion abnormalities were significantly influenced by the use of AC, in that significantly fewer abnormal and less severely abnormal segments were demonstrated in the segmental analysis as compared to NC; this was especially true for the vascular territory of the left anterior descending artery and the right coronary artery. It is concluded that AC with a moving line source is feasible in patients with all degrees of probability of CAD. AC has a significant impact on the assessment of the severity and extent of myocardial ischaemia, especially in the posterior and septal wall.
Mitochondrial changes are frequently observed in muscle fibers of patients with inclusion body myositis (IBM) and polymyositis (PM), suggesting that mitochondrial function may be especially impaired in these forms of inflammatory myopathies. Intranuclear and cytoplasmic tubulofilamentous inclusions are characteristic, although not totally specific for IBM. In the present cases, the inclusions were strikingly pleomorphic when chloroquine had been given for long periods. The nuclear inclusions were always tubular, whereas the cytoplasmic filaments had either a tubular, a helical, or a cross-striated structure with different diameters and arrangements in association with myelin-like figures, and vacuoles. Abnormal mitochondria containing paracrystalline, globoid, and other inclusions, noted in IBM, were occasionally also seen in PM or vasculitis. By contrast, in the latter, no intranuclear or cytoplasmic tubulofilamentous inclusions were apparent in muscle fibers. This study reports for the first time the presence of membrane-bound crystalloid inclusions in a muscle fiber with numerous abnormal mitochondria; similar structures have thus far only been observed in macrophages. The identity and function of these inclusions remains unknown. Using PCR analysis we detected different mtDNA deletions not only in IBM, but also in PM and vasculitis, indicating at least some degree of association between the structural mitochondrial abnormalities and the mtDNA mutations. There was no topographical correlation between the presence of tubular or helical filaments and the mitochondrial abnormalities. As already noted by others, the mitochondrial changes in IBM were more frequent than expected in this age group. It is suggested that the presence of the mtDNA deletions in IBM and PM are not primary, but rather the result of the underlying, presumably immunological disorder causing nuclear and secondary or simultaneous mitochondrial changes.
Electrical coupling is a widespread feature of developing neuronal circuits and it contributes to the generation of patterned activity. In the developing rat hippocampus, release of GABA by coactive hilar interneurones generates widespread synchronized activity. Here it is shown that hilar interneurones strongly rectify in the outward direction when depolarized. This depolarization-induced hyperpolarization, abolished by gap junction uncouplers, is modulated by nitric oxide. This phenomenon might represent a current-shunting mechanism of the excess current by providing functional inhibition at a developmental stage when GABA is excitatory. Spatial buffering of the current might represent an osmotic mechanism for growth and differentiation.
In order to reassess the role of nerve growth factor (NGF) on rat basal forebrain cholinergic neurons (BFCNs) survival and/or phenotype maturation during the early postnatal life, we immunoneutralized NGF in vivo. Hybridoma cells producing the neutralizing anti-NGF monoclonal antibody alphaD11 were implanted in the lateral ventricle of the rat at different postnatal ages (P2, P8 and P15) and the effects on the number and the soma size of cholinacetyltransferase (ChAT) positive neurons were analysed 1, 2 or 3 weeks after the injection. A marked decrease in the number and in the soma size of BFCNs was observed implanting hybridoma cells at P2 and performing the analysis 1 week later. These effects are reversed 3 weeks after the implant of hybridoma cells at P2. At this time point, the levels of alphaD11 antibodies in the brain parenchyma are still in a vast molar excess over endogenous NGF. No effects on BFCNs were observed implanting alphaD11 cells at P15 while LGN neurons showed marked shrinkage. Our results demonstrate that the reduction in the number of ChAT-positive neurons during the first two postnatal weeks of anti-NGF treatment is not due to cell death. We conclude that NGF is not a survival factor for BFCNs, and that the influence of NGF on BFCNs cell maturation during the first 2 postnatal weeks is transient and reversible. Our results on tyrosine kinase (Trk) coexpression, suggest that NGF may cooperate with other factors in the cholinergic phenotype differentiation and maintenance after the second postnatal week.
Epidemiology of thyroid diseases in iodine-sufficient areas (ISA) deals with sporadic goiter, thyroid autoimmune diseases, and thyroid cancer. A comparison between the different studies performed is difficult because methods have changed over time and selection criteria and definitions such as prevalence or incidence were not used consistently by some authors. Sporadic goiter: in ISA, autoimmune processes play a major role in the development of sporadic goiter. In adults, sporadic diffuse goiter is most frequent in young women (16%), perhaps due to additional relative iodine deficiency especially in pregnancy, and declines with age (<10%). Sporadic nodular goiter increases from 5% in young women to 9% in older women. Autoimmune thyroid disease (AITD): thyroid autoantibodies (TAb) and histopathological lymphocytic infiltration of the thyroid is much more common in ISA (4.6% in women; 1.1% in men) than in iodine-deficient areas (IDA). The prevalence and incidence of hypothyroidism and hyperthyroidism varies, depending on whether overt and subclinical forms are included and whether newly or previously diagnosed dysfunction is considered. In an overview of the literature, the prevalence is 2 in 1000 for overt and 6 in 1000 for subclinical hyperthyroidism in ISA. The values for hypothyroidism are 5 in 1000 and 15 in 1000, respectively. Change from IDA to ISA: in former IDA, the percentage of hyperthyroidism increases up to 4 years after salt iodination. Whereas this effect is transient for Plummer's disease, a change from IDA to ISA seems to lead to a permanent increase in overt and subclinical Graves' disease. Thyroid cancer: most studies demonstrate that the histopathological types of thyroid cancer are different in IDA and ISA. There is a tendency toward an increase in differentiated and decrease of anaplastic cancer. The ratio of papillary to follicular thyroid cancer ranges from 6.5:1 to 3.4:1 in areas with high iodine intake, decreases 3.7:1 to 1.6:1 in areas with moderate iodine intake, and ranges from 1.7:1 to 0.19:1 in IDA.
We report a case of a 48-yr-old woman who underwent surgery because of papillary oxyphilic thyroid carcinoma pT3. After total thyroidectomy, we administered 2960 MBq (131)I for ablation of the residual tissue. initial follow-up visits showed no clinical, radiological or scintigraphic evidence of residual or metastatic thyroid tissue. Serum thyroglobulin levels (Tg) and (131)I whole-body scintigraphy were negative. Three years after thyroidectomy, the patient experienced seizures, and as a consequence a brain tumor was removed. It was an undetected metastasis of the primary thyroid carcinoma. Histological examinations showed that neither the primary tumor nor the metastasis produced any Tg. With this fact in mind and the knowledge of negative (131)I whole-body scans we had to concentrate on radiological (CT and MRI scans) and nonspecific scintigraphic methods such as 201TI whole-body scintigraphy in our management of the patient. Further follow-up demonstrated multiple metastasis by 201TI whole-body scan (mediastinum, bones and soft tissue), and most of them have been removed by surgery. This case report demonstrates that, in addition to (131)I whole-body scans and measurement of serum Tg, the use of nonspecific tracers like 201TI is important to detect (131)I and/or Tg negative metastases.
The establishment of synaptic connections and their refinement during development require neural activity. Increasing evidence suggests that spontaneous bursts of neural activity within an immature network are mediated by gamma-aminobutyric acid via a paradoxical excitatory action. Our data show that in the developing hippocampus such synchronous burst activity is generated in the hilar region by transiently coupled cells. These cells have been identified as neuronal elements because they fire action potentials and they are not positive for the glial fibrillary acidic protein staining. Oscillations in hilar cells are "paced" by a hyperpolarization-activated current, with properties of Ih. Coactivated interneurons synchronously release GABA, which via its excitatory action may serve a neurotrophic function during the refinement of hippocampal circuitry.
Cholinergic neurones of basal forebrain (BF) express receptors for nerve growth factor (NGF) and are sensitive to NGF. In many CNS structures, including the BF region, the expression of NGF and its receptors is developmentally regulated. To test whether BF neurones depend on NGF during restricted time windows of postnatal development, we antagonized endogenous NGF by implanting, in the lateral ventricle of the rat, hybridoma cells producing blocking antibodies specific for NGF. Implants were performed at postnatal day 2 (P2), P8 and P15. BF cholinergic neurones were drastically reduced in number only when endogenous NGF was antagonized during the first postnatal week. We conclude that BF cholinergic neurones are sensitive to NGF deprivation only during early postnatal development.
Technetium-99m tetrofosmin, a lipophilic cationic complex molecule, was introduced for myocardial imaging. In some biodistribution studies it has also been reported to accumulate in the thyroid gland. Our objectives were to determine which thyroid nodules retain tetrofosmin and whether preoperative evaluation of malignancy is possible. Tetrofosmin scintigraphy was performed in 57 patients with a cold thyroid nodule on previously performed pertechnetate scintigraphy. All patients had undergone ultrasonography and sonographically guided fine-needle aspiration biopsy. The tetrofosmin scintigrams were obtained 5 min (early image) and 1 h (late image) after intravenous injection of 370 MBq. Only nodules that showed clear tracer retention after 1 h in comparison with retention at 5 min were classified as TETRO positive. Nodules without late retention were classified as TETRO negative. All patients underwent surgery and the histological results were compared with the results of tetrofosmin scintigraphy. Ten out of 11 patients with thyroid carcinoma (two pT1, three pT2, five pT4) were TETRO negative. One patient with papillary carcinoma (pT2) was TETRO positive. The mean nodular to thyroid tissue (N/T) ratio for the late scan was 1.0+/-0.20. There were 21 patients with thyroid adenomas (seven follicular, seven microfollicular and seven oxyphilic); 15 of these patients were TETRO positive and six TETRO negative. The mean N/T ratio for the late images was 1.34+/-0.41. All patients with degenerative goitre (24 cases) and the one patient with Hashimoto's disease were TETRO negative after 1 h and the N/T ratio was 0.92+/-0.12 on the late scan. Our results indicate that 99mTc-tetrofosmin scanning is of little value preoperatively in distinguishing thyroid carcinoma from other thyroid nodules. Tetrofosmin tends to demonstrate thyroid adenomas but does not have a routine role in the assessment of thyroid nodules.
Breast cancer, the most common malignancy in women, still poses a challenge to diagnostic procedures and therapy. Despite low specificity routine mammography is the method of choice to screen women for breast cancer. In the last years other additional diagnostic procedures such as high frequency ultrasonography (US) and especially magnetic resonance imaging (MRI) have improved breast cancer diagnosis. However, all these imaging methods are lacking in specificity which makes biopsy or surgery necessary. The purpose of our study was to evaluate prospectively the sensitivity, specificity, PPV and NPV of scintimammography with a new cationic complex Tc-99m tetrofosmin in patients with suspicious mammographic lesions. One hundred and thirty seven patients in whom mammography and/or high resolution ultrasonography (10 MHz) revealed suspicious breast lesions were studied with Tc-99m tetrofosmin scintimammography. In 84 of them biopsy and/or surgery was performed for histological evaluation. After intravenous injection of 555 MBq Tc-99m tetrofosmin planar images in anterior and lateral projections (5 min.p.i.) and SPECT imaging including 3-D-reconstruction (20 min.p.i.) were performed. Scintimammography was evaluated as negative, equivocal (+), probably (+2) or definitely (+3) positive. Planar scintimammography with Tc-99m tetrofosmin was negative in 46 patients (43 true negative-f.n; 3 false negative-f.n.) and positive in 38 patients (27 true positive-t.p.; 11 false positive-t.p.). Using SPECT imaging Tc-99m scintimammography was negative in 43 cases (41 t.n.; 2f.n.) and positive in 41 cases (28 t.p.; 13 t.p). Sensitivity of Tc-99m tetrofosmin scintimammography in this prospective study was 90%, specificity 80%, PPV 71% and NPV 93% for planar imaging and 93%, 76%, 68% and 95% for SPECT, respectively. Scintimammographic results in patients with suspicious breast lesion show, that Tc-99m tetrofosmin accumulates in breast cancer as well as in some fibroadenoma with high cellularity. However, the high NPV of 93 and 95% respectively excludes breast cancer in suspicious mammographic lesions to a very high degree and therefore reduces the need of biopsy and/or surgery in most of these patients.
We have identified a cluster of mitochondrial tRNA(Leu[UUR]), mutations in a severe case of infantile myopathy. There were A to G transitions found at mtDNA positions 3259, 3261, 3266 and 3268. These point mutations change the anticodon arm and the anticodon UAA, normally found in tRNA(Leu[UUR]), to UGA which is the one of the tRNAs(Ser[UCN]). This is the first anticodon alteration described in this tRNA. Another swap straight to the anticodon of tRNA(Pro) alone was recently described in a less severe case. Until now infantile myopathies have not been attributed to defined mtDNA alterations. This study reports for the first time mtDNA point mutations causing this early onset of a mitochondrial disorder. The apparent homoplasmy of these mutations and especially the location in the anticodon must be considered lethal, if the child would not have been respirated for 5 years from its birth.
Computer retrieval in a database, comprising 7,225 muscle cases, revealed that mitochondrial myopathies do not occur more frequently in inflammatory myopathies (3.74%) than in the whole series (3.69%). A more detailed study of inclusion body myositis (IBM), however, showed that severe mitochondrial alterations were apparent in about twice as many IBM cases as expected. This confirms recent studies of others although a causal relationship has thus far not been established. Identification of mitochondrial deletions by Southern blotting corresponded to the presence of severe structural abnormalities of mitochondria. Peripheral neuropathy of variable severity was noted in all cases of IBM and mitochondrial myopathy. By contrast, the association of severe mitochondrial abnormalities with polymyositis, systemic scleroderma, and vasculitis observed in some cases of the present series may be incidental or age dependent.
The uptake of 99mTc-tetrofosmin in enlarged lymph nodes, of the lung hilus, in the case of sarcoidosis Stage I (histopathologically confirmed by mediastinoscopic biopsy) is demonstrated. On a routine chest radiograph of a 78-yr-old woman, hilar lymphadenopathy was first detected. In the following mammography, disseminated micro calcifications were found in the left breast and a 99mTc-tetrofosmin study was performed for detection of breast cancer. Scintigraphy using 99mTc-tetrofosmin showed clear uptake in the hilar lymph nodes, but not in the left breast. The 99mTc-tetrofosmin uptake in the hilar lymph nodes was due to sarcoidosis confirmed by histology. Therefore, 99mTc-tetrofosmin scintigraphy may be useful in patients with suspected sarcoidosis, especially in Stage I.
OBJECTIVE: The aim of the study was to evaluate the influence of human anti-mouse antibodies on the measurement of thyrotropin. INVESTIGATIONS: Samples from 11 patients with measureable human anti-mouse antibody titres (19 micrograms/l-3880 micrograms/l) after radioimmuno-scintigraphy were analysed with 6 different thyrotropin immuno-radiometric assay kits (IRMA). Each sample was analysed in the routine way (sample influenced by human anti-mouse antibodies), as well as after incubation with murine immunoglobulin to precipitate human anti-mouse antibodies (samples not influenced by human anti-mouse antibodies). RESULTS: Two kits showed clear deviations of measured thyrotropin levels when the human anti-mouse antibody titres were higher than 1350 micrograms/l. A third kit was influenced to a lesser extent by human anti-mouse antibodies. Three of the 6 investigated thyrotropin IRMA kits produced thyrotropin values that were unaffected by the presence of elevated human anti-mouse antibodies. In comparison with former studies after immunotherapy, the thyrotropin deviations were marginal. However, differences were found between the commercially available thyrotropin assays. According to the results of this study only three out of the six investigated kits were unaffected by human anti-mouse antibodies. CONCLUSION: Since thyrotropin is one of the key quantities for the endocrinologist dealing with the thyroid gland, every laboratory should ensure high quality thyrotropin assays by critically analysing their method for human anti-mouse antibody.
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