Ultrastructure of lymphocytes from patients with paracoccidioidomycosis in the lymphocyte transformation test by phytohemagglutinin.

The morphology and ultrastructure of peripheral blood lymphocytes from patients with paracoccidioidomycosis (PCM) and from unaffected individuals (controls) were studied before and after Ficoll-Hypaque separation and at the end of culture, stimulated with phytohemagglutinin. Patient lymphocytes were cultured in medium with autologous plasma (from the patient himself) and with homologous plasma (from an unaffected donor), while donor lymphocytes were cultured in medium with plasma from a patient or with plasma from the donor himself. The Ficoll-Hypaque mixture caused no morphological or ultrastructural changes in the lymphocytes of patients or of unaffected donors. Patient lymphocytes cultured in medium with autologous plasma showed different degrees of cytoplasmic and nuclear alterations, such as organelle dissolution, vacuoles, amorphous masses, deformed nuclei, and absence of nucleoli. Lymphocytes from control individuals cultured in patient plasma also showed ultrastructural alterations, though they were less marked, and a reduced number of 'blasts'. Patient lymphocytes cultured in medium with homologous plasma (from a control individual) showed a morphology similar to that of lymphocytes from control individuals cultured in medium with their own plasma, although with a lower number of 'blasts'. On the basis of the results obtained using that methodology, we draw the following conclusions: separation by Ficoll-Hypaque does not seem to alter the ultrastructure of patient or donor lymphocytes; patients with diffuse PCM and more markedly impaired general condition can exhibit lymphocytes with morphological and ultrastructural alterations capable of affecting their biological systems and functionality.(ABSTRACT TRUNCATED AT 250 WORDS)

The use of decalcified granulated homologous cortical bone matrix in the correction of diaphyseal bone defect. An experimental study in rabbits.

In order to investigate an alternative way to correct diaphysial bone defects, granulated decalcified homologous cortical bone was used as a graft. Because of the suitable anatomic arrangement, the forearm of rabbits was chosen as an experimental model of bone defect. A 2 cm long bone cylinder was removed from the diaphysis of both radii preserving the periosteum. The artificial bone defect was filled with granular decalcified bone on the right side. The left side was used as control and kept empty or filled with undecalcified granular bone. The 18 animals were sacrificed in batches, 3,6, and 9 weeks after the operation. New bone formation was followed by X-rays, routine histology and incorporation of calcein blue, xylenol orange and tetracycline. In the decalcified granular bone grafts new bone formation was already detected at the first week and 9 weeks after the graft operation there was a well developed cylindric ossicle, in 89% of the cases. In some cases a medullary canal was present. No bone formation was found neither in the empty defects nor in the ones filled with undecalcified granular bone grafts.