Patterns of liana community diversity and structure in a tropical rainforest reserve,Ghana: effects of human disturbance

Most studies have concluded that liana diversity and structure increase with disturbance. However, a contradictory pattern has emerged recently calling for more research in the area. Liana diversity and structure were investigated in three forest types that differ in disturbance intensity (nondisturbed, moderately disturbed and heavily disturbed forest: NDF, MDF and HDF, respectively) in the Atewa Range Forest Reserve, Ghana. In each forest type, 10 square plots of 0.25 ha were demarcated. Lianas with diameter ≥1 cm located on trees with diameter ≥10 cm were enumerated. A total of 429 individuals representing 40 species, 29 genera and seventeen families were identified in the study. Shannon diversity and species richness of lianas were significantly lower in the HDF (P < 0.05). Liana density and basal area differed significantly across all forest types (P < 0.0001). The importance value index (IVI) of most liana species varied greatly across the forest types. The current study has provided evidence to support the pattern of decreasing liana diversity and structure with disturbance in some tropical forests. Further studies are recommended to gain more understanding of the factors that are responsible for the divergent liana responses to disturbance in tropical forests.     

Reducing boll weevil populations by clipping terminal buds and removing abscised fruiting bodies

Cotton pests damaging fruiting bodies (squares and young bolls) are difficult to control and their damage results in direct yield loss. Small growers, with low technological inputs, represent a large portion of cotton growers worldwide comprising more than 76 countries; they rely mainly on cultural practices to counteract pest attack in their crops. Boll weevil, Anthonomus grandis Boheman (Coleoptera: Curculionidae), oviposition involves puncturing cotton squares and young bolls, causing abscission. We examined the impact on boll weevil population of collecting abscised cotton fruiting bodies and clipping plant terminals at 50% boll maturation in the field during two cotton‐growing seasons and under field cage conditions. Greatest numbers of damaged squares occurred ca. 117 days after planting and clipped plants resulted in reduction of abscised structures and adult boll weevils compared with non‐clipped plants, irrespective of cotton variety. Damaged young bolls were found ca. 128 days after planting in 2009 and 2011, but clipping had no effect. Numbers of boll weevils found in plants of the varieties BRS 201 and BRS Rubi (both in 2009) and BRS Rubi (in 2011) were, respectively, 13‐, 17‐, and 20‐fold greater when clipping plus collecting abscised fruiting bodies were not practiced. Furthermore, the average percentage of the boll weevil parasitoid Bracon vulgaris Ashmead (Hymenoptera: Braconidae) emerging from abscised and collected structures was similar between clipped and non‐clipped plant terminals in both seasons. Clipping plant terminals did not result in yield reduction and reduced adult boll weevil production. Collecting abscised reproductive structures, clipping plant terminals, and using both practices together reduced boll weevil populations by as much as 63, 57, and 79%, respectively, in cage trials. Thus, these practices cause significant impact on boll weevil populations and are feasible of adoption, especially for smallholder cotton growers.     

Soulamarin Isolated from Calophyllum brasiliense (Clusiaceae) Induces Plasma Membrane Permeabilization of Trypanosoma cruzi and Mytochondrial Dysfunction

Chagas disease is caused by the parasitic protozoan Trypanosoma cruzi. It has high mortality as well as morbidity rates and usually affects the poorer sections of the population. The development of new, less harmful and more effective drugs is a promising research target, since current standard treatments are highly toxic and administered for long periods. Fractioning of methanol (MeOH) extract of the stem bark of Calophyllum brasiliense (Clusiaceae) resulted in the isolation of the coumarin soulamarin, which was characterized by one- and two-dimensional ¹H- and ¹³C NMR spectroscopy as well as ESI mass spectrometry. All data obtained were consistent with a structure of 6-hydroxy-4-propyl-5-(3-hydroxy-2-methyl-1-oxobutyl)-6″,6″-dimethylpyrane-[2″,3″:8,7]-benzopyran-2-one for soulamarin. Colorimetric MTT assays showed that soulamarin induces trypanocidal effects, and is also active against trypomastigotes. Hemolytic activity tests showed that soulamarin is unable to induce any observable damage to erythrocytes (c_max. = 1,300 µM). The lethal action of soulamarin against T. cruzi was investigated by using amino(4-(6-(amino(iminio)methyl)-1H-indol-2-yl)phenyl)methaniminium chloride (SYTOX Green and 1H,5H,11H,15H-Xantheno[2,3,4-ij:5,6,7-i′j′]diquinolizin-18-ium, 9-[4-(chloromethyl)phenyl]-2,3,6,7,12,13,16,17-octahydro-chloride (MitoTracker Red) as fluorimetric probes. With the former, soulamarin showed dose-dependent permeability of the plasma membrane, relative to fully permeable Triton X-100-treated parasites. Spectrofluorimetric and fluorescence microscopy with the latter revealed that soulamarin also induced a strong depolarization (ca. 97%) of the mitochondrial membrane potential. These data demonstrate that the lethal action of soulamarin towards T. cruzi involves damages to the plasma membrane of the parasite and mitochondrial dysfunction without the additional generation of reactive oxygen species, which may have also contributed to the death of the parasites. Considering the unique mitochondrion of T. cruzi, secondary metabolites of plants affecting the bioenergetic system as soulamarin may contribute as scaffolds for the design of novel and selective drug candidates for neglected diseases, mainly Chagas disease.     

Diagnosis of cystic pancreatic lesions by cytologic examination and carcinoembryonic antigen and amylase assays of cyst contents.

The contents obtained by fine needle aspiration (FNA) from 41 pancreatic cysts in 32 patients were studied cytologically and assayed for amylase and carcinoembryonic antigen (CEA) levels, which have been shown to discriminate pancreatic pseudocysts from mucinous cystic neoplasms and necrotic cystic carcinomas. The results were correlated with the histopathologic findings following surgery or with a clinical and radiologic follow-up of up to two years. The clinical, radiologic and cytologic characteristics did not discriminate pseudocysts from cystic neoplasms. The amylase content of cysts was high in pseudocysts, cystic carcinomas and mucinous cystic neoplasms. The mean CEA content was highest in cystic carcinomas and mucinous cysts and low in pseudocysts. The cytologic diagnosis of mucinous cystic neoplasms and carcinomas had a sensitivity of 54% and a specificity of 91%. The diagnosis of these lesions based on a CEA level greater than 10 ng/ml had a sensitivity of 100% and a specificity of 81%. The adjunctive use of CEA content analysis enhanced the sensitivity of the cytologic diagnosis of mucinous cystic neoplasms and carcinomas to 100%.     

EPR studies of chlorocatechol 1,2-dioxygenase: evidences of iron reduction during catalysis and of the binding of amphipatic molecules

Chlorocatechol 1,2-dioxygenase from Pseudomonas putida (Pp 1,2-CCD) is a dioxygenase responsible for ring cleavage during the degradation of recalcitrant aromatic compounds. We determined the zero-field splitting of the Fe(III) cofactor (|D| = 1.3 +/- 0.2 cm(-1)) by electron paramagnetic resonance (EPR) experiments that along with other structural data allowed us to infer the Fe(III) coordination environment. The EPR spectrum of the ion shows a significantly decrease of the g = 4.3 resonance upon substrate binding. This result is rationalized in terms of a mechanism previously proposed, where catechol substrate is activated by Fe(III), yielding an exchange-coupled Fe(II)-semiquinone (pair). The Pp 1,2-CCD capacity of binding amphipatic molecules and the effects of such binding on protein activity are also investigated. EPR spectra of spin labels show a protein-bound component, which was characterized by means of spectral simulations. Our results indicate that Pp 1,2-CCD is able to bind amphipatic molecules in a channel with the headgroup pointing outwards into the solvent, whereas the carbon chain is held inside the tunnel. Protein assays show that the enzyme activity is significantly lowered in the presence of stearic-acid molecules. The role of the binding of those molecules as an enzyme activity modulator is discussed.     

Helianthus tuberosus agglutinin directly induces neutrophil migration, which can be modulated/inhibited by resident mast cells.

We investigated the effect of Helianthus tuberosus agglutinin (HTA) on neutrophil migration in vivo and in vitro. The role of resident cells in this effect was analyzed. Peritonitis was induced by injecting stimuli into rat (150-200 g) peritoneal cavities, and in vitro neutrophil chemotaxis was performed using a Boyden microchamber. HTA (80, 200, or 500 microg/mL per cavity) induced significant in vivo neutrophil migration (p < 0.05); in vitro assays showed that this lectin also induced neutrophil chemotaxis, an effect inhibited by the incubation of lectin associated with alpha-D(+)-mannose, its specific binding sugar. Depletion of the resident-cell population by peritoneal lavage did not alter HTA-induced neutrophil migration (200 microg/mL per cavity). The opposite strategy, increasing peritoneal macrophages by intraperitoneally injecting rats with thioglycollate, did not enhance the neutrophil migration produced by HTA (200 microg/mL per cavity). In addition, injection of supernatant from HTA-stimulated macrophage culture (300 microg/mL) into rat peritoneal cavities did not induce neutrophil migration. However, reduction of the peritoneal mast-cell population potentiated the neutrophil migration (p < 0.05) induced by HTA (200 microg/mL per cavity). Lectin from H. tuberosus has a direct neutrophil chemotatic effect that is modulated by mast cells.     

Inhibition of CD73 improves B cell-mediated anti-tumor immunity in a mouse model of melanoma

CD73 is a cell surface enzyme that suppresses T cell-mediated immune responses by producing extracellular adenosine. Growing evidence suggests that targeting CD73 in cancer may be useful for an effective therapeutic outcome. In this study, we demonstrate that administration of a specific CD73 inhibitor, adenosine 5'-(α,β-methylene)diphosphate (APCP), to melanoma-bearing mice induced a significant tumor regression by promoting the release of Th1- and Th17-associated cytokines in the tumor microenvironment. CD8(+) T cells were increased in melanoma tissue of APCP-treated mice. Accordingly, in nude mice APCP failed to reduce tumor growth. Importantly, we observed that after APCP administration, the presence of B cells in the melanoma tissue was greater than that observed in control mice. This was associated with production of IgG2b within the melanoma. Depletion of CD20(+) B cells partially blocked the anti-tumor effect of APCP and significantly reduced the production of IgG2b induced by APCP, implying a critical role for B cells in the anti-tumor activity of APCP. Our results also suggest that APCP could influence B cell activity to produce IgG through IL-17A, which significantly increased in the tumor tissue of APCP-treated mice. In support of this, we found that in melanoma-bearing mice receiving anti-IL-17A mAb, the anti-tumor effect of APCP was ablated. This correlated with a reduced capacity of APCP-treated mice to mount an effective immune response against melanoma, as neutralization of this cytokine significantly affected both the CD8(+) T cell- and B cell-mediated responses. In conclusion, we demonstrate that both T cells and B cells play a pivotal role in the APCP-induced anti-tumor immune response.     

Persistence of experimental Rocio virus infection in the golden hamster (Mesocricetus auratus)

Rocio virus (ROCV) is an encephalitic flavivirus endemic to Brazil. Experimental flavivirus infections have previously demonstrated a persistent infection and, in this study, we investigated the persistence of ROCV infection in golden hamsters (Mesocricetus auratus). The hamsters were infected intraperitoneally with 9.8 LD50/0.02 mL of ROCV and later anaesthetised and sacrificed at various time points over a 120-day period to collect of blood, urine and organ samples. The viral titres were quantified by real-time-polymerase chain reaction (qRT-PCR). The specimens were used to infect Vero cells and ROCV antigens in the cells were detected by immunefluorescence assay. The levels of antibodies were determined by the haemagglutination inhibition technique. A histopathological examination was performed on the tissues by staining with haematoxylin-eosin and detecting viral antigens by immunohistochemistry (IHC). ROCV induced a strong immune response and was pathogenic in hamsters through neuroinvasion. ROCV was recovered from Vero cells exposed to samples from the viscera, brain, blood, serum and urine and was detected by qRT-PCR in the brain, liver and blood for three months after infection. ROCV induced histopathological changes and the expression of viral antigens, which were detected by IHC in the liver, kidney, lung and brain up to four months after infection. These findings show that ROCV is pathogenic to golden hamsters and has the capacity to cause persistent infection in animals after intraperitoneal infection.