Ultrastructural identification of oligodendrocyte/myelin proteins in corpus callosum of hypothyroid animals

Thyroid hormone (T3) deficiency impairs the development of the CNS, particularly myelination. We have previously described an increase in the frequency of morphological abnormalities in the central myelin sheath in a hypothyroidism model, which reinforced the hypothesis of a role for T3 in myelin compaction. However, there are no data concerning the cellular distribution of myelin proteins in hypothyroid animals. In the present work, we describe the distribution of 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase), myelin basic protein (MBP) and proteolipid protein (PLP) throughout the central myelin sheath of a hypothyroidism model. We used euthyroid and hypothyroid adult rats at 90 days of age. In order to induce hypothyroid status, animals received 0.02% methimazol from the 19th gestation day onwards. After perfusion with a fixative mixture, small pieces of corpus callosum were obtained, dehydrated and embedded in LR White resin. Ultrathin sections were immunoreacted, using specific antibodies revealed by a secondary antibody coupled to colloidal gold particles of 10 nm. Gold particle density per region of myelin sheath for each one of these proteins was obtained. In normal animals, CNPase, PLP and MBP were identified in sites that had already been described in previous studies. In hypothyroid animals, CNPase was identified in the region corresponding to compact lamellae, which normally does not contain this protein, while, in this same region, PLP and MBP immunolabeling were decreased. These results suggest that thyroid hormone deficiency impairs the distribution of the major oligodendrocyte/myelin markers. This effect may justify the reduction in myelin sheath compaction previously demonstrated in a similar model of hypothyroidism.     

Acute Antibody-Mediated Rejection Following Heart Transplantation

Acute antibody-mediated rejection (AMR) in heart transplantation is often associated with hemodynamic compromise, and is associated with increased mortality and development of accelerated transplant coronary artery disease (TCAD). The diagnosis of AMR has historically been controversial and outcomes with aggressive immunosuppressive therapy including plasmapheresis and cyclophosphamide are poor. Advances in diagnostic techniques like the demonstration of immunopathologic evidence for antibody-mediated rejection by deposition of the complement split product C4d in tissue and detection of anti-HLA antibodies by flow cytometry will assist in further characterizing AMR. Immunosuppression targeting B-lymphocytes and use of m-TOR inhibitors to alter the predilection to develop TCAD and improve survival in AMR remains to be proven.     

ASSESSMENT OF THE PREVALENCE INDEX ON SIGNS OF COMBINATION SYNDROME IN PATIENTS TREATED AT BAURU SCHOOL OF DENTISTRY,UNIVERSITY OF SAO PAULO

A group of destructive changes occurring in jaws in patients with maxillary complete dentures and mandibular removable partial dentures (bilaterally) has been described in the literature as the combination syndrome. However, this condition is not clinically observed in all patients. The aim of this study was to establish the prevalence index on signs of combination syndrome and to verify whether these changes also occurred in patients rehabilitated with a mandibular removable partial denture (unilaterally). Sample was composed of 44 patients, completely edentulous in the maxilla. Thirty-two patients had a Kennedy Class I removable partial denture and 12 a Kennedy Class II. Three major alterations were observed in 20.5% of the studied population. Nevertheless, these changes were present only in 25% of patients with Kennedy Class I removable partial denture. Based on the findings of this study, it can be concluded that patients with Kennedy Class II removable partial denture do not have similar signs that lead to the combination syndrome’s condition.     

P64Shoe contact dermatitis

Because of its numerous etiologies, foot dermatitis can be difficult to diagnose despite of thorough history and physical examination. The differential diagnoses are challenging and include allergic contact dermatitis, dyshidrosis, juvenile plantar dermatosis, atopic dermatitis, lichen planus.
Methods:  In the Cutaneous Allergology Unit of the Department of Dermatovenereology of Pedro Hispano Hospital, between 1999 and 2003, 804 patients were observed with suspicion of contact dermatitis. In 9,3% there was a history of recurrent foot dermatitis. All these patients were patch tested with GPEDC standard and shoe series and shoe fragments.
Results:  Twenty‐five patients revealed shoe contact dermatitis with positive reactions to the series tested, with an average age of 40 years. The dermatitis involved the dorsal aspect of the foot in 20 patients and the volar aspect in 8, the lateral aspects in 4, typically sparing the instep and flexural creases of the toes. The most common allergen were para‐tertiary‐butylphenol formaldehyde resin – PTBFR (12 patients‐48%), mercapto mix (7 patients‐28%), potassium dichromate (24%), mercaptobenzothiazole (24%).
Conclusions:  The most common causes of shoe contact dermatitis were glues, followed by rubber components and chromated leather. As in other studies the most common allergen in shoe dermatitis in Portugal is PTBFR in neoprene adhesives. Rubber components (accelerators) were the second more frequent allergen in this study, probably a direct result of improved fixation of chrome and a change in footwear style and chrome sensitivity explains leather allergy. Other causes of foot contact dermatitis are iatrogenic complication, clothing (socks), cosmetics, adhesive tape and professional etiology.     

Development of a transformation system for Crinipellis perniciosa,the causal agent of witches' broom in cocoa plants

Protoplasts of the pathogenic plant fungus, Crinipellis perniciosa, were transformed to hygromycin B resistance using the pAN7-1 plasmid, which contains the Escherichia coli hph gene under the control of Aspergillus nidulans regulatory sequences. The pAN7-1 plasmid was introduced by PEG/CaCl(2) treatment. Transformation frequencies of 1.6-2.5 transformants/microg of DNA were achieved. About 54% of the transformants were abortive and 40 analyzed transformants were mitotically stable and showed different hygromycin B resistance levels. The presence of the hph gene was checked by PCR in five transformants and the integration of multiple plasmid copies into different genome sites was observed by Southern analysis. This is the first report of a C. perniciosa transformation system and represents an important step for further research into genetic manipulation of this fungal plant pathogen.     

Changes in Avidity and Level of Immunoglobulin G Antibodies to Mycobacterium tuberculosis in Sera of Patients Undergoing Treatment for Pulmonary Tuberculosis

Much is known about specific antibodies and their titers in patients with tuberculosis. However, little is known about the avidity of these antibodies or whether changes in avidity occur during the progression of the disease or during treatment. The aims of this study were to determine the avidity of antibodies to Mycobacterium tuberculosis in patients with pulmonary tuberculosis, to explore the value of avidity determination for the diagnosis of tuberculosis, and to study changes in levels of antibodies and their avidity during treatment. Antibody avidity was measured by an enzyme-linked immunosorbent assay with thiocyanate elution. Avidity indices and serum levels of immunoglobulin G to M. tuberculosis were determined for 22 patients with pulmonary tuberculosis before and during treatment and for 24 patients with other pulmonary diseases. Antibody levels and avidity were both significantly higher in untreated tuberculosis patients than in the controls. Avidity determination had more diagnostic potential than determination of the antibody levels. Tuberculosis patients with a long duration of symptoms had higher antibody avidity than those with a recent onset of symptoms, indicating affinity maturation of specific antibodies during active disease. In the early phase of treatment, a decrease in antibody avidity was observed for 73% of all tuberculosis patients, accompanied by an initial increase in antibody levels in 36% of these patients. These phenomena could be explained by an intense stimulation of the humoral response by antigens released from killed bacteria, reflecting early bactericidal activity of antituberculous drugs leading to the production of low-affinity antibodies against these released antigens.     

Genomic analysis of the nuclear receptor family: new insights into structure, regulation, and evolution from the rat genome

Completion of the Rattus norvegicus genome sequence enabled a global inventory and analysis of the nuclear receptors (NRs) in three mammalian species. Forty-nine NR members were found in mouse, 48 in human. Forty-seven were found in the rat, with gaps at the locations expected for the other two. Pairwise comparisons of their distribution in rat, mouse, and human identified 11 syntenic NR gene blocks, including three small clusters of two or three closely related genes, each spanning 40 kb to 1700 kb. The exon structure of the ligand-binding domain suggests that exon shuffling has played a role in the evolution of this family. An invariant splice junction in all members of the NR family except LXRbeta suggests a functional role for the intron. The ligand-binding domains of PXR and CAR are among the most divergent in the family. Their higher nucleotide substitution rates may be related to the central role played by these two NRs in the metabolism of the foreign compounds and may have resulted from limited positive selection.     

Schistosoma mansoni: impairment of the cell-mediated immune response in mice.

Skin-graft rejection in mice experimentally infected with Schistosoma mansoni is delayed when grafting is performed 60 days after the infection. In mice infected 30 days prior to the grafting, the grafts were rejected at the same time both in infected and in control animals. This observation indicates that impairment of cell-mediated immune response occurs in mice with mature S. mansoni infections.     

A mutation in the ectodomain of herpes simplex virus 1 glycoprotein B causes defective processing and retention in the endoplasmic reticulum.

Herpes simplex virus 1 (HSV-1) glycoprotein B (gB) is one of several envelope glycoproteins required for virion infectivity and is the only one known to oligomerize into homodimers. To study the conformational constraints for translocation of HSV-1 gB to the surface of eukaryotic cells, we analyzed the transport through the exocytic pathway of the wild-type glycoprotein and of mutant forms with insertions in the ectodomain and intracellular carboxy terminus. Transient expression of the glycoproteins in COS-1 cells showed that an insertion at position 479 in the amino-terminal ectodomain of gB, shown previously by reactions with monoclonal antibodies to have altered the conformation of the molecule, also had a drastic effect on transport, precluding exit of the mutant from the endoplasmic reticulum (ER) and transport to the Golgi and the plasma membrane. The fact that the mutant, gB-(Lk479), formed dimers suggests that local changes in assembled regions caused the transport defect. Mutants containing insertions at residues 600 of the ectodomain and 810 in the intracellular domain were slightly retarded in their rate of transport from the ER to the Golgi. The glucose-regulated proteins GRP78 and GRP94, which are resident proteins of the ER, associated with partially glycosylated, faster-migrating forms of gB but not with the fully processed, more slowly migrating product. GRP78 and GRP94 formed complexes with the mutant gB-(Lk479), which was degraded in the ER. Our results indicate that GRP78, and perhaps also GRP94, acts as a chaperone in the assembly of native gB oligomers and also binds to aberrant forms of the molecule, arresting their transport from the ER and possibly serving as markers for protein degradation in this compartment of the exocytic pathway.