Efficacy of a Cladosporium sp. fungus against Helicoverpa armigera (Lepidoptera: Noctuidae), other insect pests and beneficial insects of cotton

A strain of the fungus Cladosporium sp. (RM16) from an egg of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) was assessed as a potential biocontrol agent for this pest. Pathogenicity of the fungus was tested against H. armigera eggs and larvae, cotton aphids (Aphis gossypii Glover; Homoptera: Aphididae), and silverleaf whitefly type B (Bemisia tabaci Gennadius; Hemiptera: Aleyrodidae). The pathogenicity of the fungus to the predatory red and blue beetles (Dicranolaius bellulus Guérin-Méneville; Coleoptera: Melyridae), transverse ladybird beetles (Coccinella transversalis Fabricius; Coleoptera: Coccinellidae), green lacewings (Mallada signatus Schneider; Neuroptera: Chrysopidae) and damsel bugs (Nabis kinbergii Reuter; Hemiptera: Nabidae), was also assessed in the laboratory. Fungus treatment resulted in failure to hatch of up to 64% of H. armigera eggs (compared with 11% in the controls) and mortality of 54% of first instar H. armigera larvae (compared with 5% in the controls). In contrast, it was not pathogenic to later instar H. armigera larvae. Cladosporium RM16 was also efficacious against the sap-sucking insect pests of cotton that were tested. No significant harmful effect of the fungus was found on any of the four beneficial predatory insects assessed in this study. Cladosporium RM16 has the potential as biological control agent to support integrated pest management in cotton farming systems, although this needs intensive study.     

The genomic ancestry of individuals from different geographical regions of Brazil is more uniform than expected

Based on pre-DNA racial/color methodology, clinical and pharmacological trials have traditionally considered the different geographical regions of Brazil as being very heterogeneous. We wished to ascertain how such diversity of regional color categories correlated with ancestry. Using a panel of 40 validated ancestry-informative insertion-deletion DNA polymorphisms we estimated individually the European, African and Amerindian ancestry components of 934 self-categorized White, Brown or Black Brazilians from the four most populous regions of the Country. We unraveled great ancestral diversity between and within the different regions. Especially, color categories in the northern part of Brazil diverged significantly in their ancestry proportions from their counterparts in the southern part of the Country, indicating that diverse regional semantics were being used in the self-classification as White, Brown or Black. To circumvent these regional subjective differences in color perception, we estimated the general ancestry proportions of each of the four regions in a form independent of color considerations. For that, we multiplied the proportions of a given ancestry in a given color category by the official census information about the proportion of that color category in the specific region, to arrive at a "total ancestry" estimate. Once such a calculation was performed, there emerged a much higher level of uniformity than previously expected. In all regions studied, the European ancestry was predominant, with proportions ranging from 60.6% in the Northeast to 77.7% in the South. We propose that the immigration of six million Europeans to Brazil in the 19th and 20th centuries--a phenomenon described and intended as the "whitening of Brazil"--is in large part responsible for dissipating previous ancestry dissimilarities that reflected region-specific population histories. These findings, of both clinical and sociological importance for Brazil, should also be relevant to other countries with ancestrally admixed populations.     

Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR

Background

Studies examining the association between telomere length and cancer risk have often relied on measurement of telomere length from a single blood draw using a real-time PCR technique. We examined the reliability of telomere length measurement using sequential samples collected over a 9-month period.

Methods and Findings

Relative telomere length in peripheral blood was estimated using a single tube monochrome multiplex quantitative PCR assay in blood DNA samples from 27 non-pregnant adult women (aged 35 to 74 years) collected in 7 visits over a 9-month period. A linear mixed model was used to estimate the components of variance for telomere length measurements attributed to variation among women and variation between time points within women. Mean telomere length measurement at any single visit was not significantly different from the average of 7 visits. Plates had a significant systematic influence on telomere length measurements, although measurements between different plates were highly correlated. After controlling for plate effects, 64% of the remaining variance was estimated to be accounted for by variance due to subject. Variance explained by time of visit within a subject was minor, contributing 5% of the remaining variance.

Conclusion

Our data demonstrate good short-term reliability of telomere length measurement using blood from a single draw. However, the existence of technical variability, particularly plate effects, reinforces the need for technical replicates and balancing of case and control samples across plates.     

Monokine induced by interferon gamma and IFN-gamma response to a fusion protein of Mycobacterium tuberculosis ESAT-6 and CFP-10 in Brazilian tuberculosis patients

IFN-gamma responses to Mycobacterium tuberculosis antigens ESAT-6 and CFP-10 have been proposed as specific markers of M. tuberculosis infection. Monokine induced by gamma interferon (MIG/CXCL9) has been shown to be expressed by IFN-gamma stimulated mononuclear cells and to attract activated T-cells through the chemokine receptor CXCR3. Since MIG is induced early in the response to IFN-gamma, measuring MIG may provide an interesting marker to assess downstream IFN-gamma induced responses, in contrast to assays that mainly focus on quantifying production of IFN-gamma per se. We, therefore, investigated MIG and IFN-gamma responses to a fusion protein of ESAT-6 and CFP-10, and compared responses to the conserved mycobacterial antigen 85B (Ag85B) and purified protein derivative (PPD) of M. tuberculosis, in 29 BCG vaccine controls and 24 TB patients. IFN-gamma secreting cells were determined by ELISPOT, and MIG production was measured by ELISA and flow cytometry. Production of MIG in response to ESAT-6/CFP-10, Ag85B and PPD correlated overall with increased numbers of IFN-gamma secreting cells (r=0.55, P<0.0001). A significant increase was noted among patients compared to controls in the secretion of IFN-gamma and MIG following stimulation with ESAT-6/CFP-10 or PPD (P<0.05). Moreover, MIG intracellular expression was higher in TB patients compared to BCG vaccines (P<0.05) in response to ESAT-6/CFP-10 or PPD. We conclude that MIG production correlates significantly with enhanced T-cell IFN-gamma production induced by M. tuberculosis-specific antigens ESAT-6/CFP-10. These results point to MIG as a potential novel biomarker that may be helpful in assessing downstream responses induced by IFN-gamma in TB.     

Preliminar evaluation of cytokines in the hepatitis C-schistosomiasis co-infection

Evaluation of hepatic fibrosis is usually performed by histopathological examination of biopsies. However, this is an invasive and potentially dangerous procedure. Several studies have proposed serum biological markers of hepatic fibrosis. This communication evaluates the use of serum cytokines as markers of hepatic fibrosis in hepatitis C, schistosomiasis, and co-infection.     

Analogues of SB-203207 as inhibitors of tRNA synthetases

SB-203207 and 10 analogues have been prepared, by elaboration of altemicidin, and evaluated as inhibitors of isoleucyl, leucyl and valyl tRNA synthetases (IRS, LRS, and VRS, respectively). Substituting the isoleucine residue of SB-203207 with leucine and valine increased the potency of inhibition of LRS and VRS, respectively. The leucine derivative showed low level antibacterial activity, while several of the compounds inhibited IRS from Staphylococcus aureus WCUH29 more strongly than rat liver IRS.     

Gonads of males and females of the mangrove tree crab Aratus pisonii (Grapsidae: Brachyura: Decapoda): a histological and histochemical view

Nicolau, C.F., Nascimento, A.A., Machado‐Santos, C., Sales, A. and Oshiro, L.M.Y. 2011. Gonads of males and females of the mangrove tree crab Aratus pisonii (Grapsidae: Brachyura: Decapoda): a histological and histochemical view. —Acta Zoologica (Stockholm) 00 :1–9. This study describes the microscopic anatomy of the male and female gonads and the spermatogenesis and oogenesis of the mangrove tree crab Aratus pisonii. Males and females were captured in a mangrove marsh in Guaratiba (23°04′S, 44°10′W), Rio de Janeiro State, Brazil. The testes are composed of spermatogonia I (7.82 ± 0.84 μm), spermatogonia II (6.12 ± 0.72 μm), spermatocytes I (5.62 ± 0.71 μm), spermatocytes II (5.00 ± 0.42 μm), spermatids (4.01 ± 0.33 μm), and spermatozoa (2.58 ± 0.18 μm). The spermatozoids are sent to the vas deferens, which is divided into three regions: anterior vas deferens, middle vas deferens and posterior vas deferens. There are no indications of development as the production of male gametes was continuous throughout the study period. In the females, there are four ovary development stages: previtellogenesis, early‐stage vitellogenesis, mature vitellogenesis, and postspawning. Five types of cells were found in the gonads: oogonia (5.23 ± 1.31 μm), oocytes in early development (19.84 ± 5.16 μm), previtellogenic oocytes (49.49 ± 6.87 μm), vitellogenic oocytes (87.51 ± 10.23 μm), and mature oocytes (174.78 ± 29.46 μm). The findings of this study indicate that A. pisonii females lay eggs on multiple occasions throughout the study period.     

Microbial control of cotton pests. Part I: Use of the naturally occurring entomopathogenic fungus Aspergillus sp. (BC 639) in the management of Creontiades dilutus (Stal) (Hemiptera: Miridae) and beneficial insects on transgenic cotton crops

The development and adoption of transgenic (Bt) crops that express the Bacillus thuringiensis (Bt) toxin has reduced the use of synthetic insecticide on transgenic crops to target Helicoverpa spp., the major insect pest of cotton in Australia. However, it has also increased the threat posed by sucking pests, particularly Creontiades dilutus (green mirid), which are unaffected by the Bt toxins in transgenic cotton crops. Here we report the efficacy of the entomopathogenic fungus Aspergillus sp. (BC 639) in controlling the infestation of transgenic cotton crops by C. dilutus and promoting interactions of transgenic cotton with beneficial insects. The results showed that the number of C. dilutus adults and nymphs recorded on plots treated with 1000, 750, 500, 250 ml/ha BC 639 fungus formulation were the same as on plots treated with the recommended concentration of the commercial insecticide Fipronil. The fungus was found to have minimal effect on predatory insects compared with Fipronil and was most effective against C. dilutus when applied at the rate of 500 ml/ha (equivalent to 50 g spores/ha). At this rate, the fungus was as effective as Fipronil for controlling C. dilutus populations and ensured the survival of predatory beetles, lacewings and spiders compared with Fipronil treatment. The yield from fungus-treated plots was 5.24 bales per acre compared with 5.40 and 3.88 bales per acre for Fipronil-treated and unsprayed plots, respectively. The ability of the BC 639 strain to control C. dilutus infestations of transgenic cotton crops while conserving beneficial insect populations suggests its potential for supplementing integrated pest management programs to reduce the use of synthetic insecticides for transgenic cropping systems.     

The cloning, expression, purification, characterization and modeled structure of Caulobacter crescentus β-Xylosidase I

The xynB1 gene (CCNA 01040) of Caulobacter crescentus that encodes a bifunctional enzyme containing the conserved β-Xylosidase and α-L: -Arabinofuranosidase (β-Xyl I-α-L: -Ara) domains was amplified by PCR and cloned into the vector pJet1.2Blunt. The xynB1 gene was subcloned into the vector pPROEX-hta that produces a histidine-fused translation product. The overexpression of recombinant β-Xyl I-α-L: -Ara was induced with IPTG in BL21 (DE3) and the resulting intracellular protein was purified with pre-packaged nickel-Sepharose columns. The recombinant β-Xyl I-α-L: -Ara exhibited a specific β-Xylosidase I activity of 1.25?U?mg(-1) to oNPX and a specific α-L: -Arabinofuranosidase activity of 0.47?U?mg(-1) to pNPA. The predominant activity of the recombinant enzyme was its β-Xylosidase I activity, and the enzymatic characterization was focused on it. The β-Xylosidase I activity was high over the pH range 3-10, with maximal activity at pH 6. The enzyme activity was optimal at 45?°C, and a high degree of stability was verified over 240?min at this temperature. Moreover, β-Xylosidase activity was inhibited in the presence of the metals Zn(2+) and Cu(2+), and the enzyme exhibited K(M) and V(Max) values of 2.89?±?0.13?mM and 1.4?±?0.04?μM?min(-1) to oNPX, respectively. The modeled structure of β-xylosidase I showed that its active site is highly conserved compared with other structures of the GH43 family. The increase in the number of contact residues responsible for maintaining the dimeric structure indicates that this dimer is more stable than the tetramer form.