Comparative analysis of monoclonal antibodies against prostate-specific membrane antigen (PSMA).

BACKGROUND: Prostate-specific membrane antigen (PSMA), also known as glutamate carboxypeptidase II (GCPII), is generally recognized as a diagnostic and therapeutic cancer antigen and a molecular address for targeted imaging and drug delivery studies. Due to its significance in cancer research, numerous monoclonal antibodies (mAbs) against GCPII have been described and marketed in the past decades. Unfortunately, some of these mAbs are poorly characterized, which might lead to their inappropriate use and misinterpretation of the acquired results. METHODS: We collected the 13 most frequently used mAbs against GCPII and quantitatively characterized their binding to GCPII by enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). Using a peptide library, we mapped epitopes recognized by a given mAb. Finally, we assessed the applicability of these mAbs to routine experimental setups, including Western blotting, immunohistochemistry, and flow cytometry. RESULTS: ELISA and SPR analyses revealed that mAbs J591, J415, D2B, 107-1A4, GCP-05, and 2G7 bind preferentially to GCPII in native form, while mAbs YPSMA-1, YPSMA-2, GCP-02, GCP-04, and 3E6 bind solely to denatured GCPII. mAbs 24.4E6 and 7E11-C5.3 recognize both forms of GCPII. Additionally, we determined that GCP-02 and 3E6 cross-react with mouse GCPII, while GCP-04 recognizes GCPII and GCPIII proteins from both human and mouse. CONCLUSION: This comparative analysis provides the first detailed quantitative characterization of the most commonly used mAbs against GCPII and can serve as a guideline for the scientific community to use them in a proper and efficient way.

Relationship between ALS and the degree of cognitive impairment, markers of neurodegeneration and predictors for poor outcome. A prospective study.

INTRODUCTION: Amyotrophic lateral sclerosis (ALS) is a common neurodegenerative disease affecting motor neurons and may be associated with impaired cognition. Reliable prognostic factors for ALS patients are still missing. METHODS: We prospectively included 67 patients, 42 women and 25 men, with clinically defined ALS. The disease severity was assessed and the patients underwent SPECT, lumbar puncture with determination of tau, hyperphosporylated tau (p-tau) and beta-amyloid and a detailed neuropsychological assessment using a standardized test battery. In patients who died, a detailed neuropathologic evaluation was performed. RESULTS: The mean survival duration was 26.8 months. The delay between the first signs and confirmation of the diagnosis was 12.75 months. Cognitive impairment did not have an impact on the evolution of the disease. There was no correlation between neuropsychological and SPECT findings. Higher age at onset, more pronounced handicap and elevated beta-amyloid in the CSF were associated with shorter survival times. In brain tissue from nine of the deceased patients with ALS and dementia, all showed signs of comorbidity, six had hallmarks of frontotemporal lobar degeneration (FTLD) and three showed Alzheimer disease pathology. Brain tissues form 11 deceased ALS patients who did not show signs of dementia, had only changes compatible with a diagnosis of motor neuron disease. CONCLUSION: In our prospective study, age, disease severity and CSF beta-amyloid levels taken together were a risk factor suggesting shorter survival times. Dementia is relatively frequent in ALS and may be a consequence of either FTLD or result from co-existing Alzheimer disease.

[Malignant subtype of cystosarcoma phyllodes with brain metastases]. / Maligní forma cystosarcoma phyllodes s metastázami do mozku.

Cystosarcoma phyllodes is an uncommon type of breast tumour. Benign, borderline, and malignant subtypes have been described. Central nervous system metastases of the malignant subtype of cystosarcoma phyllodes are rare and associated with poor prognosis. We report on a patient with malignant cystosarcoma phyllodes who developed metastatic disease six years after resection of the primary breast tumour. Partial regression of a brain metastasis was achieved using radiotherapy but the patient later died due to widespread metastatic disease which was uncontrollable by systemic chemotherapy. Because metastatic malignant cystosarcoma phyllodes are largely resistant to treatment, the most important objective is to provide optimal management of the primary tumour before dissemination occurs.

[Immunohistochemical detection of TTF-1 in intraoperative bioptic samples of adenocarcinoma of the lung: a year-long experience]. / Imunohistochemický prukaz TTF-1 v peroperacních Bioptických vzorcích plicních adenokarcinomu: rocní zkusenosti.

The thyroid transcription factor 1 (TTF-1) is a highly sensitive and specific marker of adenocarcinomas of pulmonary origin in differential diagnosis of solitary pulmonary nodules. Positivity of TTF-1 as a marker of primary pulmonary tumor could have a very high impact on surgical treatment strategy. From known protocols we developed the method of immunohistochemical investigation of intraoperative bioptic samples from frozen section lasting about 15 minutes. During last year, we applied this method on 30 cases of intraoperative bioptic samples. All investigated cases were verified by immunohistochemical examination from formalin-fixed and paraffin-embedded definite tissue samples. With the exception of two samples in which the result was inconclusive from both frozen and fixed tissue, all other samples revealed the same result. Our experience demonstrates that intraoperative immunohistochemical investigation of TTF-1 in proper consequences could be a very useful tool for routine practice.

Retrospective sequence analysis of the human PRNP gene from the formaldehyde-fixed paraffin-embedded tissues: report of two cases of Creutzfeldt-Jakob disease.

The definitive diagnosis of the CJD (Creutzfeldt-Jakob disease; very rare neurodegenerative disorder) can be established only on the basis of post-mortem examination of the central nervous system tissue. Formaldehyde-fixed paraffin-embedded (FFPE) tissue samples may thus constitute the only material available for molecular pathology analyses. We performed post-mortem analysis of the coding region of the prion-protein gene (PRNP)-sequence variations in two definite CJD cases suggestive of genetic form. Only FFPE tissues were available for molecular analyses. The PRNP gene open reading frame was amplified from the genomic DNA (FFPE isolated) in four overlapping, two round semi-nested PCR products that were directly sequenced. We found known pathogenic sequence variation g.532 G>A (Asp178Asn) in patient 1 but we did not find any pathogenic sequence variation in patient 2 despite her origin from the Slovak Orava region. Based on these results, we were able to discriminate between genetic and sporadic form of CJD in patient 1 and 2, respectively. The established method was found to be efficient for the sequence-variation analysis of the entire PRNP gene coding region using the genomic DNA isolated from the FFPE tissues; it can be employed in other retrospective molecular studies.

Argyrophilic grain disease: case report of the first two cases in the Czech Republic and review of the literature.

Argyrophilic grain disease (AgD) is a relatively newly described neurodegenerative disease with late-onset dementia. Morphologically it is characterized by the presence of abundant spindle-shaped argyrophilic grains (ArG) in neuronal processes and coiled bodies in oligodendrocytes. ArG consist of abnormally hyperphosphorylated form of tau protein. AgD is a substrate of at least 5% of all dementia cases with increasing incidence in the old age. Here we report the cases of a 91-year-old woman and an 83-year-old man clinically diagnosed with dementia. Neuropathological, histochemical and immunohistochemical examination of the brain tissue show the changes to be compatible with a definite diagnosis of AgD. This is the first description of two cases of AgD in the Czech Republic.

[Correlation of the autopsy with the clinical diagnosis in common neoplasms in a general hospital]. / Autoptická korelace klinické diagnostiky castých zhoubných novotvaru v praxi vseobecné nemocnice.

The author correlated in a group of 1100 autopsies made in 1987-1989 the clinical and autoptic diagnosis of malignant neoplasms of the bronchi, lungs, stomach, large intestine and rectum, liver and extrahepatic biliary pathways, pancreas and kidneys. In the case of the stomach, large bowel, liver and pancreas the diagnosis moreover with the diagnosis of non-tumourous diseases of these organs. As compared with worldwide autoptic correlations, the diagnostic standard of carcinoma of the bronchi and lungs is roughly equal, in carcinoma of the liver it is better, in carcinoma of the extrahepatic biliary pathways and stomach it is worse. The diagnostic sensitivity of malignant tumours varies from 0.6 to 0.4 and thus in 40-60% of autoptic findings of the mentioned neoplasms an accurate clinical diagnosis was not established. The rate of diagnostic errors of the first type is, however, in this group of differing clinical and autoptic diagnoses lower than in diseases caused by infectious causal agents, the rate of errors of the second type is high. Comparison of the diagnosis of tumourous and non-tumourous diseases of the stomach, large bowel, pancreas and liver revealed roughly the same diagnostic standard with the exception of cirrhosis of the liver where it is lower.

[Correlation of the autopsy with the clinical diagnosis]. / Autoptická korelace klinické diagnostiky.

Since Morgagni's times (1761) the purpose of autopsy is correlation of the clinical diagnosis and therapy, assessment of the cause of death, description and the diagnosis of unknown diseases, teaching and later collection of epidemiological data. The clinical and pathological correlation involves assessment of the sensitivity and specificity of the clinical diagnosis (autoptic findings are conceived as the reference value), and assessment of the frequency and seriousness of diagnostic errors. The importance of autopsies is apparent from the fact that the sensitivity and specificity of clinical diagnoses of some common diseases improved between 1930 and 1977, in others it remained the same, in others it deteriorated. Deterioration pertains in particular to the diagnosis of infectious diseases, some types of malignant tumours and diseases of the circulation, regardless of technological advances of diagnostic methods. The rate of errors in the diagnosis of basic diseases leading to death and immediate causes of death where knowledge of the autoptic diagnosis would probably changes the therapy and prognosis varies round 10%, the frequency of similar errors, where this knowledge would not alter the above procedures is twice or three times as high. If for the interpretation of autoptic results the "epidemiological" principle is used, the different incidence of common diseases in the population, as compared with official statistics becomes apparent, the official statistics being based on the collection of data from the death certificates. The latter data are biased by a high rate of diagnostic errors and selection phenomena.

[Autopsy correlations in the clinical diagnosis of serious diseases caused by infections in the practice of a general hospital]. / Autoptické korelace klinické diagnostiky závazných onemocnení zpusobených infekcními prícinami v praxi vseobecné nemocnice.

The author correlated in a group of 1100 autopsies made in 1987-1989 the clinical and autoptic diagnosis of active tuberculosis of the lungs and organs, bacterial meningitis, acute and subacute endocarditis, septicaemia, acute cholangitis, diffuse suppurative peritonitis, renal infections, and pneumonias. Class I diagnostic errors, i.e. those where knowledge of the diagnosis before autopsy would have probably changed therapy and the prognosis, were encountered in all groups of correlated diseases, however, with a high frequency in tuberculosis of the lungs and organs, septicaemia, renal infections and pneumonias. It seems that the diagnostic vigilance to some common serious diseases caused by infections declines in the practice of general hospitals. This experience was recorded also in hospitals abroad. Causes of diagnostic errors are discussed.

[Neuropathology of the amnesia syndrome in old age]. / Neuropatologie amnestického syndromu ve stárí.

In the 8-10th decade of life, people can be found who are not demented in the sense of WHO definition, but who are amnestic ("a memory problem only"). This defect is probably a result of the lesion of recent declarative memory. Immediate and remote memory are almost intact. Neuropathology of 17 such brains found numerous plaques and tangles in amygdalas and especially tangles in the IIth and IIIth strata of the entorhinal cortex and in subicula of hippocampi in 11 out of them. The number of plaques and tangles in frontal, parietal and temporal neocortex in these cases was not significant. In two other cases no significant ó "alzheimer-like" changes were found, but there was numerical atrophy of neurons of CA 1 sector of hippocampi in their caudal half perhaps due to chronic ischaemic lesion in one of them. In the other one there was an infarction of the left isthmus collateralis and retrosplenic cortex. In four cases no unequivocal structural basis for the amnesia was found--except slight numerical atrophy of large neurons of the nucleus basalis Meynerti. No significant diencephalic pathology was found. Severe "binswanger-like" changes were found in two cases of amnesia and in one case of control group, all of them were hypertonics. There exists a possibility that a part of "isolated" and relatively "benign" amnestic defects in old people is not a result of the aging of the brain but a sign of evolution of Alzheimer's disease and/or ischaemic vascular lesion of the brain destroying or disconnecting some of its information bottlenecks.

[Correlation of autopsy findings with the clinical diagnosis in common diseases of the circulatory system in a general hospital]. / Autoptická korelace klinické diagnostiky castých onemocnení obehového systému v praxi vseobecné nemocnice.

In a group of 1100 autopsies made in 1987-1989 the author correlates the clinical and autoptic diagnosis of embolism of the pulmonary artery, acute myocardial infarction, valvular defects, chronic cor pulmonale, cardiomyopathies, cerebrovascular attacks, subarachnoid haemorrhage; the latter was compared with the diagnosis of chronic subdural haematoma. The sensitivity of the clinical diagnosis of some diseases is low, in different categories 10-80% of diseases escape the clinical diagnosis. As compared with correlations of clinical and pathological findings published abroad, the standard of diagnosis of embolism of the pulmonary artery is roughly the same (some 80% controversial clinical and autoptic diagnoses), in acute infarction it is worse (some 50% controversial diagnoses), in acute cerebrovascular attacks it is worse only when compared with some (cca 25-40% controversial diagnoses according to the categories of the international classification). Evaluation of the sensitivity and specificity of the clinical diagnosis is only orientational, more important is the frequency of diagnostic controversies with regard to severity. Errors of the first type, i.e. those where knowledge of the autoptic diagnosis before death would have most probably influenced the therapy and prognosis is in embolism of the pulmonary artery and myocardial infarction 10 and 13% resp., in the other categories errors are rare or absent, there are, however, frequent errors in the diagnosis of subdural haematoma.

[Multi-infarct dementia--a disputed disease]. / Multiinfarktová demence--diskutované onemocnení.

Isolated multiinfarction dementia is rare among general hospital demented patients who died and were subjected to necropsy (7.8%, i. e. 9 observations from 115 clinically and pathologically diagnosed syndromes of dementia during a period of more than four years). It is more frequently combined with Alzheimer' disease (18.3% of the same group). The development of multiinfarction dementia is suggested by cerebrovascular or cardiovascular disease in the case-history, similarly as hypertension, a varying course, focal neurological symptoms incl. impaired speech, gait and swallowing, a positive Hachinski score. Multiinfarction dementia and Binswanger's disease may be variants of the same process.

[Binswanger's disease--a common disease of the brain and a frequent cause and component of dementia]. / Binswangerova nemoc--bezné onemocnení mozku, castá prícina a soucást demencí.

The author demonstrates on three selected observations clinical, radiological and neuropathological symptoms of Binswanger's disease. Its cause are changes of the long medullary arterioles which supply the white matter of the brain, most frequently in conjunction with hypertension. The basis of changes is ischaemic periventricular leucoencephalopathy. The disease is, contrary to recent ideas, very frequent and is formed by a spectrum of subclinical changes to deep dementia associated with neurological symptoms. Binswanger's disease is found isolated and combined with Alzheimer's disease or with multiinfarc dementia and is together with this unit the most frequent cause of vascular dementia in adult and old age.

[Structural changes in normal aging of the brain]. / Strukturální zmeny pri normálním stárnutí mozku.

During normal ageing of the brain its weight and volume decrease significantly and the ventricles of the brain enlarge. The number of neurons of the brain stem in some areas does not decline, in the neocortex the number of neurons declines during the period between the age of 24 and 100 years by a maximum of 10-15%. As to neurotransmitting systems, the numeric atrophy during ageing affects mainly the noradrenergic locus coeruleus and the dopaminergic substantia nigra; the cholinergic n. basalis is probably not affected. The main change which takes place in neurons during ageing is their diminution. Dendritic systems of neurons undergo during ageing regressive as well as progressive changes; in some areas progressive changes predominate, in others regressive ones, in some areas both types of changes are in equilibrium. During ageing in neurons the nucleoli diminish at a varying rate. From this it is concluded that protein synthesis is altered and there is an increase of lipofuscin; both these changes take place in different ways in different neuronal populations, similarly as the diminution of Nissl's substance. The relationship of Alzheimer's disease and ageing of the brain is obscure from the pathogenetic aspect; there exist statistically significant but only quantitative differences. No evidence was provided that ageing of the brain per se is the cause of dementia.

[Epidemiologic autopsy in Alzheimer's disease]. / Epidemiologická autopsie Alzheimerovy nemoci.

BACKGROUND: The prevalence of Alzheimer's disease (AD) in the population older than than 65 yr. has been estimated in industrialized countries at approximately 10% (8.1-12.5%). AD in the Czech republic in the year 1992 in the official statistics (Institute of Health Information and Statistics, UZIS) has been diagnosed in 22 cases of deceased and 175 sick patients. METHODS AND RESULTS: The crude rate of AD diagnosed by the Khachaturyans method among 2,197 autopsies of people aged 65-99 yr. performed at Thomayer University Hospital from 1988-1992 by use of principle of "epidemiologic" autopsy was estimated in this age group at 0.7%/100,000 thousand men/year and 1.4%/100 thousand women/year. Demographically standardized death rate in AD is 285.25/100 thousand in men and 604.24/100 thousand in women. The prevalence of AD in the whole group was 7.46%. The frequency of AD steeply rises with age with the exception of the tenth decade when the increase stops. CONCLUSIONS: AD is therefore one of most frequent diseases in this country too. AD in this republic is probably not being clinically and autoptically adequately diagnosed or its diagnoses do not become part of official statistics.

[Transmissible spongiform encephalopathy]. / Prenosné spongiformní encefalopatie.

Transmissible spongiform encephalopaties (TSE) are a group of rare fatal diseases of humans and animals. Prions, small infectious proteinaceous particles, are their supposed cause. Prusiner's theory (Nobel price 1997) proves that pathogenetically active prions are conformated physiological prions but problems of pathogenesis of TSE are still open. Most important representative of human TSE is Creutzfeldt-Jakob's disease (CJD), that of animal TES is bovine spongiform encephalopathy (BSE). It seems that a new variant of CJD found in more than 14 young people in the UK is in reality human BSE. Whether it means start of an epidemy of human BSE is not known.

[Thin-needle aspiration biopsy under ultrasonic control. The examination technic]. / Aspiracní biopsie tenkou jehlou za kontroly ultrazvuku. Technika vysetrení.

Based on experience with 298 aspiration biopsies (156 from the liver, 100 from the pancreas, 42 from other organs) made under ultrasonic control, the authors present a detailed description of the examination technique. At the same time they publish their own results with a mean sensitivity of 88%, specificity of 100% and diagnostic yield of 92%.

[Epidemiologic autopsy of Binswanger's disease]. / Epidemiologická autopsie Binswangerovy nemoci.

BACKGROUND: Binswanger's disease is the most substantial part of the continuum of ischemic vascular dementia (IVD). IVD is the second most frequent cause of dementia in industrialised countries. The frequency of IVD generally, and of the Binswanger's disease especially, is due to the method of statistical data collection, in the Czech republic not known. METHODS AND RESULTS: The crude rate of Binswanger's disease diagnosed histologically among the autopsies of 132 men and 212 woman aged 60-99 yr. performed at Thomayer's University Hospital from 1. 7. 1995 to 1. 7. 1996 by use of by principle of "epidemiologic" autopsy was estimated at 7.9%. This is about a half of the crude rate of Alzheimer's disease found in the same cohort. By estimating of histological "ischemic score", which is independent on clinical data, it is possible to diagnose the Binswanger's disease with high probability. CONCLUSIONS: The sensitivity and specificity of clinical diagnosis IVD generally and of BN in particular is low. IVD/BN is one of the most frequent and consequential ailment in higher age groups. IVD/BN is preventable and curable at earlier stages of development. Knowledge and precise and timely diagnosis of IVD/BN is fundamentally important for patients.

[Molecular genetic analysis of Alzheimer's dementia in the Czech population. The APP-717 mutation in the gene for amyloid protein precursor]. / Molekulárne genetická analýza Alzheimerovy demence v Ceské populaci. Mutace (APP-717) v genu pro amyloidní proteinový prekurzor (APP).

BACKGROUND: Although locus mutations in the gene for the amyloid precursor protein were already described in patients with Alzheimer's disease, there still are some patients where this mutation was not found and no link was found with other possible genetic loci on chromosomes 14 and 19. Therefore a group of subjects with Alzheimer's disease was subjected to tests for the presence of a mutation in the APP gene (in position 717). METHODS AND RESULTS: In a selected group of subjects with Alzheimer's disease (AD) in the gene for amyloid precursor protein in position 717 mutations of its transmembraneous region are found. The authors analyzed the genome DNA of cerebral tissue of Czech subjects for the presence of this mutation by means of the polymerase chain reaction with subsequent verification by sequencing analysis. In every subject genetic analyses from cerebral areas of the frontal lobe, temporal lobe, parietal lobe and hippocampus were performed. The methods used were the polymerase chain reaction (PCR) and sequencing. From the total number of 18 subjects with confirmed Alzheimer's disease and six non-related subjects without histopathological signs of Alzheimer's disease after the age of 90 years, three sequencing changes were found in position 717 of exon 17 of the transmembranous region of the precursor of beta-4 amyloid glycoprotein. In the first case it was substitution of thymin for adenine in codon 717, in the second case substitution of cytosine for thymine, in the third case a sporadic mutation of guanine for thymine in codon 717 was found. CONCLUSIONS: It was revealed that codon 717 could be a so-called hot spot site preferred for the preferential development of mutations in codon 717 in the gene for the amyloid precursor protein (APP).

[Morphologic verification of neoplastic portal vein obstruction]. / Morfologická verifikace nádorového uzáveru portální zíly.

BACKGROUND: Tumourous invasion into the portal circulation is a frequent and typical complication of primary carcinoma of the liver. No imaging method can, however, assess unequivocally the biological nature of the vascular occlusion and rule out reliably the possibility of benign thrombosis. The objective of the present work was to evaluate the yields of percutaneous thin-needle aspiration biopsy for the verification of intraluminal growth of a tumour, which influences to a considerable extent decisions and possible subsequent therapeutic provisions. METHODS AND RESULTS: Aspiration biopsy of the thrombus was performed in 33 patients with an ultrasonographically apparent occlusion of the portal circulation associated in 29 observations with primary carcinoma of the liver and in four instances with secondaries in the liver. For puncture thin needles--0.8-0.9 mm (21-20 G) were used inserted under ultrasonographic control. The presence of tumourous cells in the portal vessels was proved in 31 patients, i.e. in 94%. An oncologically negative result of biopsy was recorded in one patient with primary carcinoma of the liver and in a female patient with secondaries. Among morphologically verified tumourous occlusions complicating primary carcinoma of the liver a multinodular variant of a tumour was found in 11 observations and a solitary form (mean size of the focus 9 cm) in 10 observations. In 7 patients the vascular occlusion and positive result of biopsy revealed the existence of a diffuse infiltrating type of tumour. In the histological structure of the tumour which was assessed in 13 primary carcinomas of the liver differentiated types of neoplasms predominated unequivocally (10 cases) over non-differentiated tumours (3 cases). CONCLUSIONS: Thin-needle aspiration biopsy is a highly sensitive method for the detection of vascular expansion of liver tumours. It can be used successfully and without risk to assess staging of the disease where necessary.