Reinhold Kleinert

p62 Is a Common Component of Cytoplasmic Inclusions in Protein Aggregation Diseases

Exposure of cells to stress, particularly oxidative stress, leads to misfolding of proteins and, if they are not refolded or degraded, to cytoplasmic protein aggregates. Protein aggregates are characteristic features of a variety of chronic toxic and degenerative diseases, such as Mallory bodies (MBs) in hepatocytes in alcoholic and non-alcoholic steatohepatitis, neurofibrillary tangles in neurons in Alzheimers, and Lewy bodies in Parkinsons disease. Using 2D gel electrophoresis and mass spectrometry, we identified p62 as a novel MB component. p62 and cytokeratins (CKs) are major MB constituents; HSP 70, HSP 25, and ubiquitinated CKs are also present. These proteins characterize MBs as a prototype of disease-associated cytoplasmic inclusions generated by stress-induced protein misfolding. As revealed by transfection of tissue culture cells overexpressed p62 did not induce aggregation of regular CK filaments but selectively bound to misfolded and ubiquitinated CKs. The general role of p62 in the cellular response to misfolded proteins was substantiated by detection of p62 in other cytoplasmic inclusions, such as neurofibrillary tangles, Lewy bodies, Rosenthal fibers, intracytoplasmic hyaline bodies in hepatocellular carcinoma, and alpha1-antitrypsin aggregates. The presence of p62 along with other stress proteins and ubiquitin in cytoplasmic inclusions indicates deposition as aggregates as a third line of defense against misfolded proteins in addition to refolding and degradation.

Cerebral oximetry in dead subjects.

Near-infrared spectroscopy is a technique used to monitor cerebral oxygenation. To validate the method, we measured regional oxygen saturation (rSo2) in the brains of 18 dead subjects (mean age, 74.4 ± 14.6 years) 19.8 ± 18.2 h (range, 1–73) after cessation of systemic circulation, and in 15 healthy probands (mean age, 34.2 ± 8.7 years) with an INVOS 3100 cerebral oximeter. The mean (±SD) rSo2 in the dead subjects was 51.0 ± 26.8% [range, 6–88%; left, 48.4 ± 28.0% (n = 21); right, 54.4 ± 25.7% (n = 16)]. The mean rSo, in the control group was 68.4 ± 5.2% (range, 60–76%; left, 68.1 ± 5.0%; right, 68.7 ± 5.6%). After removal of the brain at autopsy in five of the dead subjects, the rSo2 was 73.4 ± 13.3% (15 measurements). Six of 18 of the dead subjects had values above the lowest values found in the healthy adults (≥60%). These findings raise concerns about the validity of cerebral rSo2 data in adults obtained by the INVOS 3100 system.

The spectrum of age-associated brain abnormalities: their measurement and histopathological correlates

Magnetic resonance imaging (MRI) has dramatically increased our ability to detect morphological abnormalities in relation to aging of the brain. Among those changes are alterations of the white matter which display high signal intensity on both proton density and T2-weighted images. They may be seen in the deep and subcortical white matter or in a periventricular location. In clinically asymptomatic individuals the reported prevalence ranges from 20% to 60% for deep and subcortical white matter hyperintensities and from 15% to 94% for periventricular changes. Besides different characteristics of the populations examined these wide ranges are a consequence of quite diverse rating schemes and measurement approaches. Inadequate grading of MRI hyperintensities may also explain some of the inconsistencies in the reported associations of white matter damage with cerebrovascular risk factors or cognitive functions. Therefore development of a commonly accepted rating scheme would be desirable. Histopathologic observations could lay the basis. Hyperintense periventricular capping of the frontal horns and a smooth halo of periventricular hyperintensity have been linked to disruption of the ependymal lining, subependymal gliosis and concomitant loss of myelin. Punctate lesions in the deep and subcortical white matter corresponded to minor perivascular reduction in myelin content possibly because of a lower permeability of thickened arteriolar walls. Larger patchy and confluent hyperintensities, however, appear to indicate more extensive ischemic damage consistent with advanced microangiopathy. In parallel, newer MRI techniques may also contribute to the delineation and separation of these various types of tissue alteration.

Immunohistochemical characterization of primitive neuroectodermal tumors and their possible relationship to the stepwise ontogenetic development of the central nervous system. 1. Ontogenetic studies.

SummaryAim of the present study was to establish different immunohistochemical staining patterns for a subsequent comparison with those of primitive neuroectodermal tumor (PNET) subsets, i.e. PNET-NOS (not otherwise specified) or PNET with focal neuronal, astrocytic or ependymal differentiation, to relate neoplastic to embryonal development. Tissue of the developing central nervous system, with special emphasis on the stepwise development of the rhombencephalon, the cerebellar and the retinal anlage, from 20 different human embryos and fetuses ranging from 3 to 30 weeks of gestational age (GA) was examined. Six neuronal markers, synaptophysin, chromogranin A, neuron-specific enolase (NSE), neurofilament protein (NFP; 160 kDa, 200 kDa, 70 and 200 kDa) and six other markers, glial fibrillary acidic protein (GFAP), S-100 protein, vimentin, myoglobin, desmin, cytokeratin, were assessed immunohistochemically. GFAP and S-100 protein appeared at the 6th week of GA in primitive glial cells of the cerebellar anlage, brain stem, rhombencephalon, and developing spinal cord, together with-as first neuronal marker-chromogranin A, then NFP (70 and 200 kDa, and 160 kDa) from the 8th week onward. NSE started in the 11th week and synaptophysin not earlier than the 16th week of GA. Interestingly, the differentiation of the retinal anlage started rather late with NSE positivity beginning from the 16th week and positive reactions to synaptophysin and NFPs only from the 25th and chromogranin A from the 28th week of GA onward.

Fluorescent In Situ Hybridization on Isolated Tumor Cell Nuclei: A Sensitive Method for 1p and 19q Deletion Analysis in Paraffin-Embedded Oligodendroglial Tumor Specimens

In oligodendroglial neoplasms, losses of chromosomal material at 1p and 19q associate with chemosensitivity and prolonged survival. Thus, 1p/19q testing is increasingly proposed for use in brain tumor diagnosis and prognostic assessment. Fluorescent in situ hybridization (FISH) is a classic technique for investigation of 1p/19q status in paraffin-embedded tissues. A major limitation of this method is truncation of tumor cell nuclei complicating assessment of hybridization results. In our study, we analyzed 1p and 19q status in a series of 79 oligodendroglial neoplasms (49 oligodendrogliomas, 30 oligoastrocytomas, WHO: 57 Grade II, 22 Grade III tumors) and controls (gliotic brain tissue: n = 4, diffuse low-grade astrocytoma: n = 4) using FISH on isolated whole tumor cell nuclei, prepared as cytospin preparations, thus bypassing the problem of nuclear truncation. For interpretation of FISH results, we used consensus criteria as defined by the SIOP—Europe Neuroblastoma Study Group for analysis of peripheral neuroblastic tumors. FISH yielded interpretable results in 98.7% for 1p and 92.1% for 19q. Chromosome 1p/19q alterations comprised deletions (1p: 79.5%, 19q: 80%) and imbalances (1p: 11.5%, 19q: 12.9%). 1p aberrations were more frequent in oligodendroglioma than in oligoastrocytoma (100% versus 75.9%, P = .001). The frequency of 1p/19q alterations was not significantly different in WHO Grade II or Grade III tumors or in primary and recurrent tumors. We conclude that FISH on isolated cell nuclei, with application of the SIOP Europe Neuroblastoma consensus criteria, is a sensitive method for detection and interpretation of 1p and 19q aberrations in paraffin-embedded tissue specimens of oligodendroglial neoplasms.

Tumefactive demyelinating lesions: conventional and advanced magnetic resonance imaging:

In rare instances, demyelinating disorders present with radiological features that mimic a brain tumour. This often leads to biopsy, which-apart from carrying significant morbidity-frequently turns out as nondiagnostic or dispensable. We therefore set out to assess the contribution of repeated conventional magnetic resonance imaging (MRI), 1H-MR spectroscopy and magnetization transfer imaging in establishing a correct diagnosis of tumefactive demyelinating lesions (TDLs). We studied two females and one male, who presented with TDLs that led to brain biopsy in two cases, for up to three years. TDLs were characterized by the following features: (a) delayed or absent response to high-dose steroids together with progressive lesion growth over several weeks; (b) late or sparse enhancement, ill-defined borders, signal inhomogeneity and considerable concomitant oedema; and (c) normalization of initial increases in lipid and lactate peaks within three to four weeks, followed by persistent, marked reductions of the neuronal marker NAA and MTR values around or below 30%. These imaging characteristics reflected the histological correlate of marked demyelination in the absence of significant inflammation. MRI techniques thus appear to have the potential to establish a correct diagnosis of this subtype of TDLs. Awareness of these possibilities might obviate the need for biopsy at least in some cases in future.

MRI in tuberculous meningoencephalitis: report of four cases and review of the neuroimaging literature

SummaryThe contribution of MRI is reported in four adult patients with tuberculous meningoencephalitis (TbM) and with autopsy correlation in one. Contrast-enhanced T1-weighted MRI revealed the characteristic basal meningeal inflammation of TbM and its focal spreading into adjacent brain. Mixed and T2-weighted pulse sequences delineated a plethora of parenchymal abnormalities. Their relation to TbM was established by a close matching of the patients neurological findings, contrast enhancement or a change in lesion size. The latter accurately reflected the clinical course in all patients. It remained difficult, however, to distinguish between ischaemic and inflammatory changes, which in some locations were intermixed even histologically. From our experience and that of other groups. MRI provides more diagnostic information in TbM than CT. Moreover, MRI promises to be a useful tool for monitoring treatment response.

Bilateral Medial Medullary Infarction: Magnetic Resonance Imaging and Correlative Histopathologic Findings

Bilateral medial medullary infarction is a rare event which clinically presents with flaccid tetraplegia sparing the face, bilateral disturbance of deep sensation, hypoglossal nerve palsy and respiratory failure. We here report a patient with such symptoms in whom magnetic resonance imaging enabled the detection of signal abnormalities in the lower brainstem as soon as 9 h after onset. Results of a control study 3 weeks later correlated well with the extent of infarction that was seen at autopsy. Early lesion detection in the lower medulla by magnetic resonance imaging and the unfavorable prognosis of patients with ischemic damage at that location may provide the rationale for aggressive therapeutic strategies in such a condition.

Primary paraganglioma simulating pituitary macroadenoma: a case report and review of the literature

Abstract A 76-year-old woman presented with an intrasellar and suprasellar mass which caused deterioration of visual acuity and bitemporal visual field defects. Trans-sphenoidal and transcranial partial resection revealed a primary chemodectoma. This tumour is very rare in the sellar area, where there are normally no paraganglionic cells. We review the literature and discuss possible mechanisms for the development of this tumour.

[Corneal melting in both eyes after simultaneous corneal cross-linking in a patient with keratoconus and Down syndrome].

Corneal cross-linking is one of the options for treatment of progressive keratoconus. Following the published standards regarding indication and treatment schedules, it seems to be a highly safe and effective operation. Only a very few severe complications, such as stromal scarring and bacterial keratitis, have been reported.We describe a patient with keratoconus and Down syndrome who was treated with corneal cross-linking on both eyes simultaneously. One week after the operation he developed central corneal melting without signs of infection in his right eye, and 1 month after the operation in his left eye. Penetrating keratoplasties had to be performed on both eyes and were successful. A possible reason for the corneal melting might have been a corneal stroma that was thinner than the proposed limit of 400 µm at the centre. The published recommended standards should be met.