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1 virus Schizophrenia Electron Microscope Studies Ultrastructural Study of the Temporal Lobe and Peripheral Blood in Schizophrenic Patients Summary. Introduction. In spite of the human, social and economic impact of schizophrenia and to be considered a disease of organic origin by several evidences, there are relatively few studies about this disease using electron microscope techniques. Patients and methods. For 25 years we have made studies about the viral hypothesis of schizophrenia by means of this technique. We have studied limbic structure samples from young adult s dead schizophrenic patients, from fetuses of schizophrenic mothers and from chicken embryos experimentally inoculated with cerebrospinal fluid from schizophrenic patients. In the last ten years we have performed studies of blood and cerebrospinal fluid samples from young schizophrenic patients. We have found some alterations with the same characteristic to those observed in the central nervous system, which are compatible with: a) the viral hypothesis and b) with herpes simplex hominis type I virus, when immunoelectronmicroscopic techniques were made. Results and conclusion. The results obtained in this work can constitute a new element favoring the possible viral etiology of schizophrenia. Dr. Segundo Mesa Castillo Hospital Psiquiátrico de la Habana 24/01/2008 Psychiatric Hospital of Havana 1 24/01/ Organic character of the disease Schizophrenia Structural alterations in schizophrenic brains in an important proportion of patients that has been shown by imagenologic and morphometric studies which point out to deeper structural atrophies in the limbic system (amygdaline nucleus and hippocampus) specially in the left cerebral hemisphere. Electron microscope studies 24/01/ /01/ Schizophrenia: electron microscopy Young adults Fetuses from schizophrenic mothers Chicken embryos inoculated with CSF Blood and cerebrospinal fluid Limbic System 24/01/ /01/

2 Sections Auditory cortex Amygdala Hippocampus Schizophrenia: electron microscopy. Central nervous system findings: Nuclear bodies Membrane alterations Particles with viral morphology Blood and cerebrospinal fluid findings: Membrane alterations, vacuoles, spherical and hexagonal particles, glycogen increase, morphological alterations 24/01/ /01/ Nuclear bodies Particles with viral morphology Nuclear envelope 24/01/ /01/ núcleo Particles Intranuclear particles Central core Particles with a central core and an envelope related to membrane structures 24/01/ /01/

3 Particles with viral morphology: below, herpes simplex virus. Above (arrow) particle observed in schizophrenia. Nucleus Particles with viral morphology Immuno-electronmicroscopy The image below is from Dennis Kunkel's excellent Microscopy Science and Photography Through a Particle labeled with anti-herpes simplex hominis type I antisera 24/01/2008 Microscope 13 24/01/ Electron microscopy studies Nucleus Fetuses and chicken embryos Cytoplasm Foetus of schizophrenic mother 300 nm 24/01/ Particles. Immunoelectromicroscopy 24/01/ Particle within an erytrocyte in a brain capillar Foetus from schizophrenic mother Digital analysis of the particles within the nucleus 24/01/ Particle labeled with anti herpes simplex antibody (arrow). Digital analysis of the particle which shows a central core and a hexagonal capsule. 24/01/

4 Cytoplasm Particles and membrane structures labeled with the antibody (anti HSV-1 antibody) Antigen antibody reaction. (arrows) Particle. Antigen antibody reaction Nucleus of neuron Particle Particle (vertical arrow) related to membrane structure (oblique arrow) 24/01/ Brain sample of chicken embryo inoculated with CSF 24/01/ Cerebrospinal fluid Negative staining and immunoelectromicroscopy Cerebrospinal fluid spherical particles 200 nm 300 nm Spherical 24/01/2008 particle labeled with coloidal gold joint to anti HSV1 antibody 21 Anti 24/01/2008 HSV1 antibody coloidal gold conjugated 22 Blood. Labelling the particles with coloidal gold with anti-hsv1 antibody Blood: 24/01/2008 particles without labelling 23 24/01/

5 Blood spherical particles Coloidal gold vacuole? Platelet Negative 24/01/2008 staining 25 24/01/ Schizophrenia: Biological Test Blood biological test Macroscopic agglutination reaction Optic microscopy Fluorescence Electron microscopy Immuno-electron microscopy 24/01/ /01/ An agglutination reaction is observed in 45% of schizophrenic patients when plasma and CSF are joined together. This agglutination is formed by platelets. Plasma plasma CSF C S F Platelets AGGLUTINATION Platelet agglutination 24/01/ Optic Microscopy 24/01/

6 Platelets Immuno-fluorescence Antiherpes antibody EOSINOPHIL Agglutination reaction Electron 24/01/2008 microscopy negative staining 31 24/01/ Anti herpes simplex hominis type I antibody Platelet Immuno-electronmicroscopy negative staining and coloidal gold 24/01/ In schizophrenic patients during the critical period of the disease big vacuoles are observed in samples of the peripheral blood which are originated from platelets (arrows). These vacuoles have hexagonal particles in some ocassions. See next slides. 24/01/ Hexagonal particles Within the big vacuoles (arrows) hexagonal particles are observed. These particles are similar to those observed in the central nervous system and are also related to membrane structures. See next slide at higher magnification. 24/01/ /01/

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