Biochemical, immunological and toxicological characteristics of the crystal proteins of Bacillus thuringiensis subsp. medellin

Characterization of the insecticidal and hemolytic activity of solubilized crystal proteins of Bacillus thuringiensis (Bt) subsp. medellin (Btmed) was performed and compared to solubilized crystal proteins of isolates 1884 of B. thuringiensis subsp. israelensis (Bti) and isolate PG-14 of B. thuringiensis subsp. morrisoni (Btm). In general, at acid pH values solubilization of the Bt crystalline parasporal inclusions (CPI) was lower than at alkaline pH. The larvicidal activity demonstrated by the CPI of Btmed indicated that optimal solubilization of CPI takes place at a pH value of 11.3, in Bti at pH values from 5.03 to 11.3 and in Btm at pH values from 9.05 to 11.3. Hemolytic activity against sheep red blood cells was mainly found following extraction at pH 11.3 in all Bt strains tested. Polyacrylamide gel electrophoresis under denaturing conditions revealed that optimal solubilization of the CPI in all Bt strains takes place at the alkaline pH values from 9.05 to 11.3. An enriched preparation of Btmed crystals was obtained, solubilized and crystal proteins were separated on a size exclusion column (Sephacryl S-200). Three main protein peaks were observed on the chromatogram. The first peak had two main proteins that migrate between 90 to 100 kDa. These proteins are apparently not common to other Bt strains isolated to date. The second and third peaks obtained from the size exclusion column yielded polypeptides of 68 and 28-30 kDa, respectively. Each peak independently, showed toxicity against 1st instar Culex quinquefasciatus larvae. Interestingly, combinations of the fractions corresponding to the 68 and 30 kDa protein showed an increased toxicity. These results suggest that the 94 kDa protein is an important component of the Btmed toxins with the highest potency to kill mosquito larvae. When crystal proteins of Bti were probed with antisera raised independently against the three main protein fractions of Btmed, the only crystal protein that showed cross reaction was the 28 kDa protein. These data suggest that Btmed could be an alternative bacterium for mosquito control programs in case mosquito larval resistance emerges to Bti toxic proteins.

Effect of gibberellic acid and the biostimulant stimulate® on the initial growth of tamarind

Plant growth regulators and biostimulants have been used as an agronomic technique to optimize the production of seedlings in various crops. This study aimed to evaluate the influence of gibberellic acid and the biostimulant Stimulate® on the initial growth of tamarind (Tamarindus indica L.). The experiments were conducted in a nursery with 50% shading, in a randomized block design with five replications and five plants per plot. Thirty eight days after sowing, the leaves were sprayed seven times a day with 0.0 (control), 0.8, 1.6, 2.4 and 3.2 mL of gibberellic acid L-1 aqueous solution and with 0.0 (control), 6.0,12.0, 18.0, and 24.0 mL Stimulate® L-1 aqueous solution. Stem diameter (SD), plant height (PH), longest root length (LRL), shoot dry mass (SDM), root dry mass (RDM) and RDM:SDM ratio were evaluated ninety days after sowing. Variance and regression analysis showed that GA3 at 4% promoted plant growth (height), but had no significant effect on stem diameter, longest root length, shoot and root dry mass and the RDM:SDM ratio. On the other hand, all concentrations of Stimulate® significantly increased plant height and shoot and root dry mass of tamarind seedlings.

THE INTERNATIONAL GROWTH OF A SOCIAL BUSINESS: A CASE STUDY

ABSTRACTIn developing countries, initiatives have often been undertaken in order to fight social and environmental problems. Since the 1990s, an increase can be seen in corporate social responsibility actions, as well as increasingly strong activities by civil society organizations. Tweenty years ago, companies and civil society organizations stood wide apart from each other, with often conflicting agendas and resistance to mutual collaboration. This reality has changed significantly. Besides the phenomenon of cross-sector partnerships, we can also observe the expansion of a particular organization type, i.e., the social business, which combines two objectives that were previously seen as incompatible: financial sustainability and the generation of social value. This article aims to discuss the factors that influence the results of a social business operating in three countries: Botswana, Brazil and Jordan. The results allow understanding the challenges involved in constructing social businesses in developing countries as well as a better understanding of the very nature of those businesses, considering the social realities where they operate.

Relative growth of Uca burgersi (Crustacea, Ocypodidae) from two mangroves in the southeastern Brazilian coast

The relative growth of the fiddler crab Uca burgersi Holthuis, 1967 was analyzed in two populations from different mangroves (Ubatumirim and Cavalo rivers) in the southeastern Brazilian coast, monthly sampled from May, 2001 to April, 2002. The population from a high productivity level mangrove reaches the sexual maturity in larger sizes than the population from a less productive site. This results can be attributed to the greater amount of nutrients found in the Cavalo than in Ubatumirim mangrove.

TIMP-1 mediates the inhibitory effect of interleukin-6 on the proliferation of a hepatocarcinoma cell line in a STAT3-dependent manner

The tissue inhibitor of metalloproteinases (TIMP)-1 is a multifunctional protein which is not only an inhibitor of matrix metalloproteinases (MMPs) but also to have a possible "cytokine-like" action. Here, we first compared mRNA expression of TIMP-1 and MMP-9 in BEL-7402 (a hepatocellular carcinoma cell line), L-02 (a normal liver cell line) and QSG-7701 (a cell line derived from peripheral tissue of liver carcinoma) using real-time quantitative RT-PCR. By evaluating the variation of the MMP-9/TIMP-1 ratio as an index of reciprocal changes of the expression of the two genes, we observed that the MMP-9/TIMP-1 ratio was about 13- and 5-fold higher in BEL-7402 than in L-02 and QSG-7701, respectively. Significantly, overexpression of TIMP-1 decreased the MMP-9/TIMP-1 ratio in BEL-7402 and then inhibited the cell growth to 60% and reduced the migration to about 30%. Meanwhile, our data showed that interleukin-6 (IL-6) (100 ng/mL) could also inhibited the cell growth of BEL-7402. Further studies indicated that TIMP-1 mediated the inhibitory effect of IL-6 on BEL-7402 cell proliferation in a STAT3-dependent manner, which could further accelerate the expression of the cyclin-dependent kinase inhibitor p21. A dominant negative STAT3 mutant totally abolished IL-6-induced TIMP-1 expression and its biological functions. The present results demonstrate that TIMP-1 may be one of the mediators that regulate the inhibitory effect of IL-6 on BEL-7402 proliferation in which STAT3 signal transduction and p21 up-regulation also play important roles.

Comparative allometry growth of some marine fish digenetic trematodes

Allometric growth variation was compared for Plagioporus idoneus, Lepocreadium pegorchis, Opecoeloides furcatus, Bacciger israelensis, Aphanurus stossichi and Parahurleytrema trachinoti collected from East Mediterranean fishes. The pharynx, the oral and the ventral sucker diameters always showed a negative allometry. The other parameters tested were variable with the species. We study the effects of some environmental factors: the influence of the host species is analysed in Plagioporus idoneous, wich parasitizes Oblada melanura, Diplodus sargus and D. vulgaris and in Lepocreadium pegorchis, wich parasitizes Pagellus erythrinus, Lithognathus mormyrus and Spicara smaris; the influence of the microhabitat and the intensity of infection is analysed in Bacciger israelensis and Aphanurus stossichi, both parasites of Boops boops. We report significant differences with the host species, for the allometric growth of the testes; the effect of the microhabitat was revealed by the hindbody allometric value; no significant difference was detected in relation with the intensity of infection.

NPK fertilization on initial growth of physic nut seedlings in Quartzarenic Neossol

Balanced fertilization is important for plant growth. There is little information on physic nut (Jatropha curcas L.) and tests with the fertilization of the species are very recent. This study evaluated the initial growth of physic nut seedlings in response to NPK rates to Quartzarenic Neossol in a greenhouse and estimated P and K critical soil levels and N, P and K in shoot dry matter after 120 days of evaluation. The treatments were arranged in a randomized, fractional factorial design (4 x 4 x 4)½, totalizing 32 treatments with three replicates, 96 experimental plots and N rates (0, 75, 150 and 300 mg dm-3) as urea; P rates (0, 45, 90 and 180 mg dm-3) as triple superphosphate and K rates (0, 50, 100 and 200 mg dm-3) as potassium chloride. After 120 days, the plants were harvested and some variables evaluated: plant height, stem diameter, shoot and root dry weight, macro and micronutrient levels in plant shoots, and soil chemical properties. The physic nut seedlings responded to NPK fertilizer in the initial growth phase; the response to N was negative. The recommended P and K rates were 25 and 67 mg dm-3, respectively. The critical levels, corresponding to the recommended P rate were 13 and 74 mg dm-3 for K in soil (Mehlich-1). The N, P and K levels in the shoot dry matter of physic nut were 37.4, 2.1 and 35.7 g kg-1, respectively.

Trypanosoma cruzi: expression of antigenic component 5 among 35 laboratory clones obtained from 18 different isozymic variants

Two monoclonal antibodies anti-component 5 of Trypanosoma cruzi (I-35/115 and II-190/30) were tested in IFA and ELISA respectively against 35 T. cruzi laboratory clones. Among the 35 clones tested, 18 different isozyme patterns were detected. All clones were recognized by both monoclonal antibodies except one clone which did not react with II-190/30. These results support the universal expression of specific component 5 within the taxon T. cruzi.

Monoterpene isolated from the essential oil of Trachyspermum ammi is cytotoxic to multidrug-resistant Pseudomonas aeruginosa and Staphylococcus aureus strains

Abstract INTRODUCTION: The aim of this study was to determine whether an herbal extract containing monoterpene exhibited activity against multidrug-resistant Staphylococcus aureus and Pseudomonas aeruginosa isolated from clinical infection samples. METHODS: The essential oil of Trachyspermum ammi (L.) Sprague ex Turrill (Apiaceae) fruit was extracted by hydrodistillation. Fruit residues were treated with hydrochloric acid and re-hydrodistilled to obtain volatile compounds. Compounds in the distilled oil were identified using gas-chromatography (GC) and GC-mass spectrometry (MS). The antibiotic susceptibility of all bacterial isolates was analyzed using both the disc diffusion method and determination of the minimum inhibitory concentration (MIC). The sensitivity of antibiotic-resistant isolates to essential oil was also determined by using the disc diffusion method and MIC determination. RESULTS: Of 26 clinical isolates, 92% were multidrug-resistant (MDR). Aromatic monoterpenes (thymol, paracymene, and gamma-terpinene) were the major (90%) components of the oil. Growth of S. aureus strains was successfully inhibited by the oil, with an inhibitory zone diameter (IZD) between 30-60mm and MIC <0.02μL/mL. The oil had no antimicrobial activity against clinical isolates of P. aeruginosa; rather, it prevented pigment production in these isolates. CONCLUSIONS: This study revealed that the essential oil of Trachyspermum ammi, which contains monoterpene, has good antibacterial potency. Monoterpenes could thus be incorporated into antimicrobial ointment formulas in order to treat highly drug-resistant S. aureus infections. Our findings also underscore the utility of research on natural products in order to combat bacterial multidrug resistance.

Acute myocardial infarction in the Brazilian State of São Paulo. In-hospital deaths from 1979 to 1996 and hospital fatality from 1984 to 1998 in the public health system

OBJECTIVE: To determine the following parameters in the Brazilian State of São Paulo: 1) the percentage of deaths due to acute myocardial infarction (AMI) occurring in hospitals; 2) the percentage of deaths due to AMI occurring in public health system hospitals as compared with all in-hospital deaths due to AMI between 1979 and 1996; 3) the fatality due to AMI in public health system hospitals from 1984 to 1998. METHODS: Data were available on the Datasus Web site (the health information agency of the Brazilian Department of Health) that provided the following: a) number of deaths resulting from AMI in hospitals; b) number of deaths resulting from AMI in public health system hospitals; c) number of hospital admissions due to AMI in public health system hospitals. RESULTS: The percentage of in-hospital deaths due to AMI increased from 54.9 in 1979 to 68.6 in 1996. The percentage contribution of the public health system to total number of deaths due to AMI occurring in hospitals decreased from 22.9 in 1984 to 13.7 in 1996; fatality due to AMI occurring in public health system hospitals had an irregular evolution from 1984 to 1992 and showed a slight trend for increased frequency from 1993 to 1998. CONCLUSION: The percentage of in-hospital deaths due to AMI has been increasing. Deaths resulting from AMI in public health system hospitals have decreased when compared with the total number of deaths due to AMI in all hospitals. Fatality due to AMI in public health system hospitals did not decrease from 1992 to 1998.

Draft genome sequences of three NDM-1-producing Enterobacteriaceae species isolated from Brazil

The emergence of multidrug-resistant Enterobacteriaceae strains producing carbapenemases, such as NDM-1, has become a major public health issue due to a high dissemination capacity and limited treatment options. Here we describe the draft genome of three NDM-1-producing isolates: Providencia rettgeri(CCBH11880), Enterobacter hormaecheisubsp. oharae(CCBH10892) and Klebsiella pneumoniae(CCBH13327), isolated in Brazil. BesidesblaNDM-1, resistance genes to aminoglycosides [aadA1, aadA2,aac(6R17;)-Ib-cr] and quinolones (qnrA1,qnrB4) were observed which contributed to the multidrug resistance profile. The element ISAba125 was found associated to theblaNDM-1 gene in all strains.

Initial growth of Bauhinia variegata trees under different colored shade nets and light conditions

Bauhinia variegata and B. variegata var. candida, commonly known as orchid trees, are small sized trees widely used for urban forestry and landscaping. Adult plants grow under full sun; in Brazil, however, seedlings are generally cultivated in commercial nurseries under natural half-shading. The objective of this study was to evaluate the influence of different colored shade nets and light conditions on the initial growth of B. variegata and B. variegata var. candida. The influence of six light conditions (red net with 50% shading; blue net with 50% shading; black net with 70% shading; black net with 50% shading; black net with 30% shading; and full sun) on the initial growth of B. variegata and B. variegata var. candida were evaluated along 160 days, and growth relationships were calculated. Seedlings showed more efficiency on the use of photoassimilated compounds when grown under full sun. Such condition is the most appropriate for seedling production of B. variegata and B. variegata var. candida, contradicting what has been performed in practice.

The antioxidant action of Polypodium leucotomos extract and kojic acid: reactions with reactive oxygen species

Two natural products Polypodium leucotomos extract (PL) and kojic acid (KA) were tested for their ability to scavenge reactive oxygen species (·OH, ·O2-, H2O2, ¹O2) in phosphate buffer. Hydroxyl radicals were generated by the Fenton reaction, and the rate constants of scavenging were 1.6 x 10(9) M-1 s-1 for KA and 1.0 x 10(9) M-1 s-1 for PL, similar to that of ethanol (1.4 x 10(9) M-1 s-1). With superoxide anions generated by the xanthine/hypoxanthine system, KA and PL (0.2-1.0 mg/ml) inhibited ·O2-dependent reduction of nitroblue tetrazolium by up to 30 and 31%, respectively. In the detection of ¹O2 by rose bengal irradiation, PL at 1.0 mg/ml quenched singlet oxygen by 43% relative to azide and KA by 36%. The present study demonstrates that PL showed an antioxidant effect, scavenging three of four reactive oxygen species tested here. Unlike KA, PL did not significantly scavenge hydrogen peroxide.

The inhibitory effect of photodynamic therapy and of an anti-VCAM-1 monoclonal antibody on the in vivo growth of C6 glioma xenografts

We investigated the effect of photodynamic therapy (PDT) and of an anti-vascular cell adhesion molecule-1 (VCAM-1) monoclonal antibody on the in vivo growth of C6 glioma. Seven days after inoculation with C6 cells, adult male Wistar rats weighing 280-300 g with MRI-confirmed glioma were randomly assigned to 4 groups (N = 15 per group): PDT + VCAM-1 antibody group; PDT group; VCAM-1 antibody group; control group. Eight days after inoculation, hematoporphyrin monomethyl ether (HMME) was administered as a photosensitizer and PDT was performed at 630 nm (illumination intensity: 360 J/cm²) for 10 min. VCAM-1 antibody (50 µg/mL) was then administered (0.5 mL) through the tail vein every other day from day 8 to day 16. At day 21, 5 rats in each group were sacrificed and cancers were harvested for immunohistochemistry and Western blot assay for the detection of VCAM-1, and TUNEL assay was used to detect apoptosis. Survival and tumor volume were recorded in the remaining 10 rats in each group. In the PDT group, tumor growth was significantly suppressed (67.2%) and survival prolonged (89.3%), accompanied by an increase in apoptosis (369.5%), when compared to control. Furthermore, these changes were more pronounced in the PDT + VCAM-1 antibody group. After PDT, VCAM-1 expression was markedly increased (121.8%) and after VCAM-1 monoclonal antibody treatment, VCAM-1 expression was significantly reduced (58.2%). PDT in combination with VCAM-1 antibody can significantly inhibit the growth of C6 glioma and prolong survival. This approach may represent a promising strategy in the treatment of glioma.