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Biomedical subjects

P Mehraein

Publications and source records attributed to P Mehraein.

104 records · Page 6Linked to original sources

Persistence of Borrelia burgdorferi and histopathological alterations in experimentally infected animals. A comparison with histopathological findings in human Lyme disease.

Gerbils appear to be susceptible to infection by human isolates of Borrelia burgdorferi; we obtained 100% infection. Isolation of the B. burgdorferi from different organs six months post infection causes a generalized infection thus demonstrating that borreliae persist in these animals for a long period. Spirochetemia was present for 14 days, apparently in two intervals. The Borrelia burgdorferi specific antibody titers increased with time after infection thus indicating the persistence of spirochetes. The intraperitoneal inoculation of the B. burgdorferi to six gerbils of groups A and B induced significant histopathologic changes in most of the major organ systems and their surrounding adipose and fibrous connective tissues. The infiltrates consisted mainly of lymphocytes and histiocytes. Various numbers of plasma cells, eosinophils and high numbers of mast cells were also present. Three further animals which served as controls displayed no histological signs of inflammation in any organ system. No significant differences were noted between the histopathological findings seen in the animals of groups A and B (infected with cells from subcultures no. 25 and with no. 5, respectively). The persistence of B. burgdorferi and the high number of organs involved with slight to severe signs of inflammation in this series can be compared to persistence and to the multiorgan involvement seen in human Lyme disease. Thus gerbils can serve as suitable experimental animals to study the pathogenesis of Lyme disease and the extent of organ damage caused by B. burgdorferi.

Animals↗

Active immunization with pC protein of Borrelia burgdorferi protects gerbils against B. burgdorferi infection.

Serious infection due to Borrelia burgdorferi and the disseminated infection characteristic of the disease possess unique treatment problems. The wide and still increasing incidence of Lyme borreliosis as well as the problems in treatment call for effective prevention strategies by active immunization. Vaccination experiments were done to determine if active immunization of gerbils with recombinant OspA and pC protects against infection with strains of B. burgdorferi. Gerbils were vaccinated with recombinant OspA and pC (20 kDa protein) and challenged four weeks later with a clone (derived from B. burgdorferi strain PKo) which expresses an abundant amount of pC but only little OspA. Non-immunized gerbils challenged with the same B. burgdorferi strain were used as controls. Both groups of immunized gerbils developed antibodies against the recombinant vaccines. The pC vaccinated group was protected against infection, whereas the OspA vaccinated group showed signs of infection. The non-vaccinated group developed generalised infection. These results show that pC should be considered as a further vaccine candidate and probably needs to be combined with OspA for an efficient vaccine against B. burgdorferi.

Animals↗

A novel two-site immunoradiometric assay for beta-endorphin using nitrocellulose as solid phase.

A two-site immunoradiometric assay for the highly specific direct quantitation of nonacetylated beta h-EP in crude brain tissue samples has been developed with a detection limit of 10 fmol per well. The assay used two different antibodies with distinct specificities: a polyclonal rabbit anti-beta h-EP antibody binding between the middle portion and the C-terminal end of beta h-EP was bound to nitrocellulose membrane discs, a solid phase with a high protein binding capacity. In the following two incubation steps, the beta h-EP containing crude tissue extract--or the beta h-EP-standard--and, subsequently, the 125I-labeled monoclonal 3-E7 mouse antibody directed against the N-terminus of beta h-EP were added. Binding of beta h-EP to the solid phase antibody in the first incubation step was not affected by the addition of cross reacting opioid peptides derived from beta h-LPH up to 10 pmol per disc. Nonspecific binding of the labeled antibody to the solid phase could be lowered to 3% of total counts by the use of PBS containing nonfat dry milk as blocking solution and incubation buffer, a procedure that did not reduce maximum specific binding. Dilution studies performed with extracts sampled from the anterior hypothalamus excluded the interference of tissue factors in the assay.

Brain↗

Cytochrome c oxidase defects of the human substantia nigra in normal aging.

Based on morphological, biochemical, and molecular biologic analyses, degeneration of the dopaminergic nigrostriatal system has been reported to occur with normal aging. In the present study, the substantia nigra of 36 human brains with normal aging was investigated by means of morphometry and immunohistochemistry. The anteromedial (Am), anterointermediolateral (Ail), posteromedial (Pm), and posterolateral (Pl) nuclei of the substantia nigra were analyzed using antibodies directed against the subunits II/III of cytochrome c oxidase (COX), the complex IV of the respiratory chain. The numerical density of melanin-positive neurons with COX defects was significantly increased in the four investigated nuclei, namely Am, Ail, Pm, and Pl. These cells did not show any histologic signs of degeneration. The numerical density of melanin-positive neurons without COX defects was decreased with aging. The data of the present study indicate that complex IV defects of neurons in the substantia nigra might be one cause of neuronal dysfunction occurring during aging.

Adult↗

Evaluation of intracerebral lesions in patients with acquired immunodeficiency syndrome. Neuropathological findings and experimental data.

In this paper we present the results of post-mortem examinations of the central nervous system in 61 male patients who died with Acquired Immunodeficiency Syndrome (AIDS); it includes 23 patients with reported neurological abnormalities at the time of presentation. The analysis revealed central nervous system (CNS) neoplasms (lymphoma, Kaposi's sarcoma) and a variety of inflammatory lesions (bacterial, fungal, protozoal and viral) in 32 cases. A total of 11 patients without opportunistic infections showed significant brain abnormalities characterized by microglial nodules and/or multinucleated giant cells, changes which are probably related to infection by human immunodeficiency virus (HIV). In addition, we describes results from a series of experiments designed to define the target cell population of HIV in the brain. The expression of CD4 complex--putative receptor for HIV--was investigated using short-term cultured brain cells taken from embryonic brain anlage and from different regions of fetal brain; glioma cells were also used. Cells derived from normal embryonic and fetal brain, as well as glioma cells, were examined with respect to their susceptibility to HIV. CD4 antigen expression could be demonstrated only on glioma cells of the permanent glioma line 85HG-59 comprised of cells with properties characteristic of astrocytes. Nevertheless, normal embryonic and fetal brain cells as well as glioma cells could be infected by HIV as documented by immunocytochemical methods and southern blot analysis. HIV infected brain cells showed reduced growth rate and altered growth pattern. This study emphasizes the diversity of HIV conditioned CNS impairments, suggesting that genomic variability of HIV may result in varying cell type preference of the virus. The experimental data indicate that CD4 expression in brain cells is probably not 'conditio sine qua non' for HIV susceptibility. The alterations of HIV-infected brain cells demonstrated provide further evidence for a direct involvement of HIV in the pathogenesis of AIDS-related neurological syndromes.

Acquired Immunodeficiency Syndrome↗

Radioimmunodetection of human glioma xenografts by radiolabelled monoclonal antibodies.

Radiolabelled monoclonal antibodies (131I-MUC 8-22, 131I-MUC 2-63) were used for external scintigraphy of human glioma xenografts. To induce transplantation tumors. 5 x 10(6) cells (85HG-66) of an in vitro established human malignant astrocytoma (N66/85) were inoculated s.c. in BALB/c-nu/nu mice. The labelling of the immunoglobulins with 131iodine was carried out according to the iodogen method, the nude mice, bearing xenograft, received 30 m. 131I-labelled intact monoclonal immunoglobulins (200mCi: 7,4MBq) and the imaging was performed on days 4, 8 and 12 after the application. After 4 days, a clear tumor accumulation of iodinated MUC 2-63 antibodies recognizing surface determinants was visible. This enrichment of monoclonal antibodies (MAbs) led to a characteristic tumor presentation on day 8. Obviously, the MUC 2-63 antibodies remain in the tumor tissue for a long time, so that even on day 12 satisfactory tumor imaging is possible. On the other hand, neither with normal mouse IgG nor with MUC 8-22 antibodies - which react with intracellular structures - could a tumor localization be achieved. The result of the studies on the distribution of 131I-MUC 2-63 on day 19 was that the activity in the tumor tissue was about 4.4 times higher than in the blood and even more times higher than in solid organs.

Animals↗

Antigenic heterogeneity of human brain tumors defined by monoclonal antibodies.

The antigenic profiles of human gliomas and in vitro established cell lines were investigated using the monoclonal antibodies (MABs) MUC 8-22 and MUC 2-63. The reactivity with tissue samples and cytospin preparations obtained from 45 brain tumors was estimated by the indirect immunoperoxidase technique. In addition, computer-assisted cytofluorometry was used to quantify the intensity and distribution of antibody-binding. Various degrees of antibody-binding among and within gliomas and glioma-derived cell lines were observed. The data show that a variable percentage of cells are not labeled with the employed MABs. The spectrum of reactivity of the selected antibodies was independent of the histological grading of gliomas. However, there were significant differences in various stages of subcultivation of glioma lines. In most cases, the heterogeneity of antigen expression decreased during successive in vitro propagation of glioma cells. The extent of variation in staining intensity values differed within cell populations and reflected the antigenic heterogeneity of human brain tumors. The findings presented here suggest that the use of MABs which recognize glioma-associated antigens facilitates the objective analysis of brain tumors and is of potential value for immunohistochemical application in surgical neuropathology.

Antibodies, Monoclonal↗

Relationship between Ki-67 positive cells, growth rate and histological type of human intracranial tumors.

The proliferation rate of 40 intracranial neoplasms (30 gliomas, 1 hemangioblastoma, 3 meningiomas, 1 neurinoma and 5 brain metastases) was investigated using the monoclonal antibody Ki-67. In eleven of the gliomas recurrences could be observed, and two of them recurred for second time. In total the Ki-67 labelling indices of 53 specimens were investigated. The Ki-67 nuclear antigen was demonstrated in frozen sections by application of the appropriate monoclonal antibodies according to a modified alkaline phosphatase-antialkaline phosphatase (APAAP) technique. The proliferation rate was evaluated by cell count calculation of the staining index. Ki-67-labelled glioma cells varied from 0.2 percent in one meningioma (WHO-grade I) to 9.1 percent in one glioblastoma. In ten glioma recurrences, higher Ki-67 staining indices could be observed than in their primaries, even when the histological grading did not change substantially. In a cerebellar hemangioblastoma, a trigeminal neurinoma and two endotheliomatous meningiomas the fraction of stained nuclei was less than one percent; however, one recurrent transitional meningioma without any histological sign of malignancy showed a staining index of 2.4 percent. The staining indices of five brain metastases of different malignancies ranged from 1.5 percent in a malignant melanoma to 6.1 percent in bronchial carcinoma. In the majority of the cases examined, the percentage of Ki-67 labelled cells was in accordance with the histologic grade of the neoplasm. In general, there was a direct relationship between the number of stained nuclei and the frequency of mitoses (mitotic index) evaluated in hematoxylin-eosin stained frozen sections. Interestingly, the frequency of mitosis and stained nuclei were higher in tumor recurrences than in the primaries. The results of this study imply that immunohistological labelling of the proliferating cell fraction should become an important additional criterion to predict the biological behaviour of human nervous system neoplasms.

Adult↗

Radioimaging of experimental glioma grafts using F (ab')2-fragments of monoclonal antibodies.

Iodinated F (ab')2-fragments of monoclonal IgG2a-antibodies (McAbs) 14 AC1 raised against membrane components of an experimental rat glioma (79FR-G-41) were used for the radioimmunodetection of glioma xenografts in athymic mice. For gamma imaging, nude mice with intramuscular glioma grafts received 10-20 micrograms of 131I-labelled whole immunoglobulins and F (ab')2-fragments of the 14AC1 McAb, respectively, with an activity of 50-150 mu Ci (1.85-5.55 MBq). Scans obtained at 24, 48, 72 and 96 hours after injection demonstrated tumor accumulation of F (ab')2-fragments at 24 hours, and adequate tumor visualization without background subtraction at 48 hours. The rapid blood clearance of F (ab')2-fragments resulted in a whole body half life of 6 hours. Scintigrams using intact antibody provided comparable tumor localization at 96 hours, but also high bloodpool radioactivity and a half life period of more than 4 days. 131I-labelled normal mouse-IgG as control for non-specific uptake of immunoglobulins in the tumor revealed no tumor visualization at any time. Rapid tumor permeation, short blood clearance and a markedly reduced background radioactivity let F (ab')2-fragments appear superior to whole immunoglobulins in immunoscintigraphy, thus indicating their potential value for the improvement of radioimmunodiagnosis of glioma disease.

Animals↗

Radioimmunodetection of gliomas by administration of radiolabelled monoclonal antibodies. Experimental data.

Radiolabelled monoclonal antibodies (McAbs) raised against membrane components of an experimental rat glioma (79FR-G-41) were administered parenterally to immunodeficient mice bearing glioma grafts for tumor radioimmunodetection by external imaging. Purified McAbs (14AC1) of IgG2a isotype were labelled with Na131I (2mCi/50ml) using the Chloramin-T method. As control, for non-specific uptake of proteins in the tumor, normal mouse IgG were also iodinated. For radioimaging, nude mice bearing gliomas in the thigh muscle were injected intravenously with 15 micrograms of the 131I-McAb with an activity of approximately 150 mu Ci. Control tumor-bearing animals received the same amount of mouse 131I-IgG. Scans obtained immediately after injecting the intact 131I-14AC1 antibody and at 24, 48, 72, and 96 hours demonstrated accumulation in the tumor. The tumor was clearly visible 48 hours following injection of 131I-labelled antibody. At 96 hours after injection, the McAb showed a clearly higher uptake into the tumor as the control IgG. The biodistribution of the injected antibody was studied at 96 hours after injection following the last gamma-imaging. At this time the blood activity was still high, but the maximum activity was found in the tumor for the specific McAb. Using the 131I-14AC1 to image glioma transplants, it could be shown that grafts are permeable for the McAb. The time-course experiments administering 131I-14AC1 antibody and normal mouse 131I-IgG, demonstrated that the localization of 131I-I4AC1 antibody in glioma grafts is the result of specific antigen binding. The scintigrams using intact antibody without background subtraction provided adequate tumor visualization, but the activity in the blood was high even 96 hours after injection. More rapid clearance of blood - pool radioactivity would possibly be achieved with F(ab')2 fragments. These in vivo glioma imaging studies, together with related in vitro binding tests, indicate the potential value of monoclonal antiglioma antibodies not only for clinical tumor radioimmunodetection, but also for the evaluation of immunotherapeutic approaches to the glioma disease of man.

Animals↗

Brain involvement in generalized argyria.

Cutaneous argyria was diagnosed in a 59-year-old woman. Manic depressive psychosis developed at about the same or a short time thereafter. The patient died 6 years later from a ruptured aortic aneurysm. At autopsy silver deposits were seen in skin, mucous membranes, heart, kidney, and liver. In the central nervous system the leptomeninges and choroid plexuses contained silver granules. In addition, silver granules were visualized in the walls of many intraparenchymal vessels, particularly of the basal ganglia, hypothalamus, substantia nigra, and cerebellum. Progressive glial changes and cellular gliosis were evident in many areas of the brain. With the electron microscope the deposition of silver granules in basal membrane structures of the choroid plexus and intracerebral vasculature was amply confirmed. Furthermore, silver deposition was seen in brain parenchymal cells inside bodies of apparently lysosomal nature. The silver content of various brain regions was determined by absorption spectrophotometry.

Argyria↗

Cellular composition and distribution of gliomesenchymal nodules in the CNS of AIDS patients.

The pathology of AIDS encephalopathy and its relation to other CNS infections in AIDS was studied in 112 autopsies of AIDS patients and 14 of HIV-positive patients. Special interest was focused on gliomesenchymal nodules (GMNs), a common finding in encephalitis of various etiologies, and on giant cells. GMNs and giant cells were found in 40 patients. These 40 were used for the present study, they were further divided into five groups: I. 21 with AIDS encephalopathy; II. 3 with HIV infection but no AIDS; III. 8 with Toxoplasma encephalitis; IV. 7 with cytomegalovirus encephalitis; and V. 1 with cryptococcus encephalitis. Comparison of the morphological features of GMNs between the five groups revealed parameters which may help distinguish GMNs in HIV infection from opportunistic infections. Small GMN size, low cellular density, ill-defined demarcation from the adjacent brain paraenchyma, minimal tissue rarefaction in the area of GMNs and a strong predilection for the superficial and deep white matter were typical of AIDS encephalopathy. In contrast, opportunistic infections were usually associated with clear demarcation from surrounding brain tissue, high cellular density, strong tissue rarefaction and a predilection for gray matter. Rod cells and macrophages in great numbers were also more commonly found in the AIDS and HIV groups.

AIDS Dementia Complex↗

Immunhistological evaluation of Creutzfeldt-Jakob disease with reference to the type PrPres deposition.

German patients suspected of having Creutzfeldt-Jakob disease (CJD) and related diseases were studied pathologically. The immunohistochemical findings after hydrolytic autoclaving pretreatment sensitively detected the synaptic-type deposition of the protease-resistant isoform of the prion protein (PrPres which thus served to establish the consensus diagnosis of CJD.

Aged↗

Kaposi's sarcoma in the cerebellum of a patient with AIDS.

A case of Kaposi's sarcoma in the left cerebellar hemisphere of a patient with AIDS is presented. The lesion was not detected with imaging techniques in the patient who, during the clinical course, did not show neurological signs and symptoms. This small nodular lesion was only revealed at autopsy. This rare finding is an example of the broad spectrum of central nervous system involvement in HIV-1-infected individuals.

Acquired Immunodeficiency Syndrome↗