Anti-inflammatory intestinal activity of Combretum duarteanum Cambess. in trinitrobenzene sulfonic acid colitis model

AIM To evaluate the anti-inflammatory intestinal effect of the ethanolic extract(EtO HE) and hexane phase(HexP) obtained from the leaves of Combretum duarteanum(Cd).METHODS Inflammatory bowel disease was induced using trinitrobenzenesulfonic acid in acute and relapsed ulcerative colitis in rat models. Damage scores, and biochemical, histological and immunohistochemical parameters were evaluated. RESULTS Both Cd-Et OHE and Cd-Hex P caused significant reductions in macroscopic lesion scores and ulcerative l e sionareas. The vegetable samples inhibited myeloperoxidase increase, as well as pro-inflammatory cytokines TNF-α and IL-1β. Anti-inflammatory cytokine IL-10 also increased in animals treated with the tested plant samples. The anti-inflammatory intestinal effect is related to decreased expression of cyclooxygenase-2, proliferating cell nuclear antigen, and an increase in superoxide dismutase. CONCLUSION The data indicate anti-inflammatory intestinal activity. The effects may also involve participation of the antioxidant system and principal cytokines relating to inflammatory bowel disease.

Novel Immunogenic Epitopes in the NaPi-IIb Protein: Production of Monospecific Antibodies Using Synthetic Peptides Outlined on Isoform Specific Regions of the Type IIb Sodium-Dependent Phosphate Transporter (NaPi-IIb)

NaPi-IIb is a multiple passage protein membrane which is primarily responsible for phosphate uptake in the kidney and in the small intestine. Beyond its physiological functions, their involvement with carcinogenesis was initially described in mid-2003, due to its distinct level of expression in normal and tumor cells of the ovary. Although less common than cervical cancer, epithelial ovarian cancer is considered the most lethal gynecologic malignancy, which is mainly due to diagnosis in the advanced stages as a result of the absence of symptoms during the onset of the disease and the lack of tools for early detection. Here, we produce antibodies that are anti-synthetic peptides that are derived from the regions of second extracellular loop of NaPi-IIb, which is a non-overlapping portion of MX35 epitope. These two 15 distinct amino acid residue peptides, designated as Let#1 and Let#2, are engineered in a very thorough way to detect specific sites only in this isoform, thus excluding cross-reactivity with other carriers of the same family. The lack of immunogenicity of small peptides is surpassed by the conjugation with carrier proteins. Using immunochemical methods, we demonstrate that polyclonal antibodies that are mono-specific for the Let#1 and Let#2 peptides recognize proteins that express similar amino acid sequences during blood circulation. Additionally, using flow cytometry, we identify NaPi-IIb in NIH:OVCAR-3 cells. The clear identification of two shorter peptides on the extracellular loop of NaPi-IIb, which are far from the monoclonal antibody MX35-recognizing epitopes, adds new promising tools for ovarian cancer follow-up and staging.

Hyphal Morphology and Elongation Alterations in Aspergillus nidulans Provoked by the Diterpene Kaurenoic Acid

Kaurenoic acid (KA), a kaurane-type diterpene extracted from leaves of Mikania hirsutissima, was previously reported as an inhibitor of vascular contractility mainly by blocking extracellular Ca2+ influx. The compound is known for several other biological activities such as antiparasitic, antispasmodic and antibacterial activity. The aim of the present study was to investigate the effect of KA on Aspergillus nidulans. KA (0.3 mM) showed fungistatic activity against A. nidulans with visible hyphal elongation and morphology damage. These effects were reverted by CaCl2 addition showing that KA interferes with intracellular Ca2+ gradient in A. nidulans. This is the first report on the mechanism of action of KA involving calcium levels by altering the elongation of fungi hyphae.

Biomonitoring the <i>Vitex gardneriana</i>Shauer (Lamiaceae) Toxic Effects to Shed Light on Bioactive Compounds against a Major Coconut Pest Mite

The coconut mite, Aceria guerreronis Keifer (Acari: Eriophyidae), is a major pest of coconut plantations (Cocos nucifera L.) worldwide. Here, we conducted a bioguided phytochemical approach using toxicity and repellency bioassays of nonpolar extract and its fractions of Vitex gardneriana Schauer (Lamiaceae) leaves to this pest. Nonpolar crude extract was fractionated by column chromatography using solvents with increased polarity and binary mixtures, resulting in five semipurified groups. The biomonitoring bioassay provided active fractions and led to the isolation and characterization of the bioactive compound squalene, a biosynthetic precursor of 20-hydroxyecdysone, which plays an important role in plant defense against arthropods. The LC50 of the crude extract of V. gardneriana for A. guerreronis was estimated to be 0.185 mg·mL-1 and LC80 = 4.123 mg·mL-1. Also, the extract was highly repellent to this pest for up to 24 h. The fractions of V. gardneriana, and also squalene, caused mortality to A. guerreronis. The potential of V. gardneriana fractions/squalene as biopesticides for controlling A. guerreronis in coconut plantations is discussed herein.