Genetic Diversity of Colletotrichum spp. an Endophytic Fungi in a Medicinal Plant, Brazilian Pepper Tree.

In this study, we reported thirty-nine endophytic fungi identified as Colletotrichum spp. associated with Brazilian pepper tree or aroeira (Schinus terebinthifolius Raddi. Anacardiaceae) in Paraná state, Brazil. These endophytes were identified by morphological and molecular methods, using PCR taxon-specific with CaInt/ITS4, CgInt/ITS4, and Col1/ITS4 primers, which amplify specific bands in C. acutatum, C. gloeosporioides lato sensu, and Colletotrichum boninensis, respectively, and by DNA sequence analysis of the nrDNA internal transcribed spacer region (ITS1, 5.8S, ITS2). We also assayed the presence of dsRNA particles in Colletotrichum spp. isolates. Combining both morphological characters and molecular data, we identified the species C. gloeosporioides, C. boninense, and C. simmondsii. However, we found a high genetic variability intraspecific in C. gloeosporioides which suggests the existence of several other species. Bands of double-stranded RNA (dsRNA) were detected in three of thirty-nine isolates. Identity of these bands was confirmed by RNAse, DNAse, and S1 nuclease treatments for the isolates LGMF633, LGMF726, and LGMF729. This is the first study reporting these particles of dsRNA in C. gloeosporioides.

Chemical composition, antinociceptive and anti-inflammatory effects in rodents of the essential oil of Peperomia serpens (Sw.) Loud.

ETHNOPHARMACOLOGICAL RELEVANCE: Peperomia serpens (Piperaceae), popularly known as "carrapatinho", is an epiphyte herbaceous liana grown wild on different host trees in the Amazon rainforest. Its leaves are largely used in Brazilian folk medicine to treat inflammation, pain and asthma. AIM OF THE STUDY: This study investigated the effects of essential oil of Peperomia serpens (EOPs) in standard rodent models of pain and inflammation. MATERIALS AND METHODS: The antinociceptive activity was evaluated using chemical (acetic acid and formalin) and thermal (hot plate) models of nociception in mice whereas the anti-inflammatory activity was evaluated by carrageenan- and dextran-induced paw edema tests in rats croton oil-induced ear edema, as well as cell migration, rolling and adhesion induced by carrageenan in mice. Additionally, phytochemical analysis of the EOPs has been also performed. RESULTS: Chemical composition of the EOPs was analyzed by gas chromatography and mass spectrometry (GC/MS). Twenty-four compounds, representing 89.6% of total oil, were identified. (E)-Nerolidol (38.0%), ledol (27.1%), α-humulene (11.5%), (E)-caryophyllene (4.0%) and α-eudesmol (2.7%) were found to be the major constituents of the oil. Oral pretreatment with EOPs (62.5-500 mg/kg) significantly reduced the writhing number evoked by acetic acid injection, with an ED(50) value of 188.8 mg/kg that was used thereafter in all tests. EOPs had no significant effect on hot plate test but reduced the licking time in both phases of the formalin test, an effect that was not significantly altered by naloxone (0.4 mg/kg, s.c.). EOPs inhibited the edema formation induced by carrageenan and dextran in rats. In mice, EOPs inhibited the edema formation by croton oil as well as the leukocyte and neutrophil migration, the rolling and the adhesion of leukocytes. CONCLUSIONS: These data show for the first time that EOPs has a significant and peripheral antinociceptive effect that seems unrelated to interaction with the opioid system. EOPs also displays a significant anti-inflammatory effect in acute inflammation models. This effect seems to be related to components which inhibit the production of several inflammatory mediators. These results support the widespread use of Peperomia serpens in popular medicine to treat inflammation and pain.

Agrobacterium tumefaciens-mediated transformation of Guignardia citricarpa.

Guignardia citricarpa, the causal agent of Citrus Black Spot, was successfully transformed via Agrobacterium tumefaciens with cassettes for gfp and bar expression. Transformation is essential to understand the role of genes during interaction between plants and its pathogens. Using a binary plasmid vector based in the pPZP201BK, both germinated conidia and physically fragmented hyphae of G. citricarpa were transformed. Eight independent transformants of G. citricarpa resistant to ammonium glifosinate displayed GFP fluorescence. The majority (93.75%) of the G. citricarpa transformants was mitotically stable and contained a single T-DNA copy ectopically integrated to the chromosome. This is the first report of G. citricarpa transformation and will allow future work on virulence determinants of the fungus and possibly its control.

An anti-inflammatory lectin from Luetzelburgia auriculata seeds inhibits adhesion and rolling of leukocytes and modulates histamine and PGE2 action in acute inflammation models.

OBJECTIVES: To study and characterize the in vivo effect of the lectin from Luetzelburgia auriculata seed on acute inflammation models. METHODS: The lectin was purified from the crude saline extract by affinity chromatography on a guar-gum matrix. Native, heat-treated, and digested lectin was evaluated for anti-inflammatory activity by using peritonitis and paw edema models. The anti-inflammatory activity was characterized by intravital microscopy, nitric oxide production, and myeloperoxidase activity. RESULTS: The lectin exhibited anti-inflammatory activity (2 mg/kg) on both models, reducing local myeloperoxidase activity. Galactose or heat treatment (100 degrees C, 10 min) reduced anti-inflammatory action. Anti-inflammation involves the inhibition of adhesion and rolling of leukocytes along with augmentation of nitric oxide in serum. The lectin inhibited the edematogenic effect of histamine and prostaglandins (PGE2) but did not alter the chemoattractant effect of IL-8. CONCLUSIONS: The results indicate that this lectin is a potent anti-inflammatory molecule. Its effects engage diverse modulatory events.

[Nontoxic multinodular goiter: evaluation of recurrence rate and function after partial thyroidectomy. Retrospective analysis of 39 cases]. / Bócio multinodular atóxico: avaliação da taxa de recidiva e função após tiroidectomia parcial. Análise retrospectiva de 39 casos.

The postoperative goiter recurrence and hypophysial-thyroid function in 39 patients who underwent partial thyroidectomy for nontoxic multinodular goiter, during the 1970-1983 period, was evaluated, seeking relations among thyroid function, extension of surgery and goiter recurrence. The incidence of recurrent goiter was approximately 15%, most of these goiters having been identified ten or more years after thyroidectomy. More extensive surgery lowered the recurrence rate but increased the risk of subclinical-hypothyroidism. No statistical significant differences with regard to serum T3, T4, F T4 and TSH (basal and after TRH) of patients with and without recurrent goiter were found. Thus, the maintenance of a recurrent nontoxic goiter does not seem to depend exclusively on increased serum TSH levels. In one patient, the postoperative treatment with thyroxine did not prevent the appearance of recurrent goiter. Since most patients who underwent thyroidectomy remain euthyroid and the goiter recurs in only a small number, the routine postoperative use of thyroid hormone for the prophylaxis of multinodular goiter recurrence seems to be questionable.

Male infertility due to congenital adrenal hyperplasia: testicular biopsy findings, hormonal evaluation, and therapeutic results in three patients.

Testicular biopsy in three infertile male patients with congenital adrenal hyperplasia showed different grades of germinal epithelium maturation, and no Leydig cells in the interstitium. In two patients with severe germinal cell hypoplasia, very low serum gonadotropins showed no response to luteinizing hormone-releasing hormone stimulation and responded to long-term clomiphene citrate administration. It is hypothesized that an increase of hypothalamic estrogen production because of local androstenedione aromatization might be responsible for the luteinizing hormone-releasing hormone block observed in these patients. All patients were submitted to adrenal suppressive therapy, and in two cases a return of the fertility status was obtained.