In vitro synergic effect of ²-lapachone and isoniazid on the growth of Mycobacterium fortuitum and Mycobacterium smegmatis

Nontuberculous mycobacteria are ubiquitous and saprophytic organisms that have been implicated in a wide spectrum of diseases due to an increasing number of immunocompromised patients. The natural resistance of atypical mycobacteria to classical antituberculous drugs has encouraged research into new chemotherapeutic agents and drug combinations. The aim of this study was to determine the in vitro antimycobacterial activities of ²-lapachone alone and in combination with isoniazid against Mycobacterium fortuitum and Mycobacterium smegmatis via the Time-Kill Curve method. A 2 log10 CFU/mL reduction in the M. smegmatis culture was observed 72 h after adding ²-lapachone at its minimum inhibitory concentration. This drug sterilised the culture in 120 h. For M. fortuitum, a reduction of 1.55 log10 CFU/mL occurred in 24 h, but regrowth was seen in contact with ²-lapachone. Both microorganisms were resistant to isoniazid. Regrowth of M. fortuitum and M. smegmatis was observed at 48 h and 72 h, respectively. In combination, these two drugs had a bactericidal effect and sterilised both cultures in 96 h. These results are valuable because antibiotic-resistant bacteria are a major public health problem.

Identification of SL addition trans-splicing acceptor sites in the internal transcribed spacer I region of pre-rRNA in Leishmania (Leishmania) amazonensis

Trypanosomatidae is a family of early branching eukaryotes harbouring a distinctive repertoire of gene expression strategies. Functional mature messenger RNA is generated via the trans-splicing and polyadenylation processing of constitutively transcribed polycistronic units. Recently, trans-splicing of pre-small subunit ribosomal RNA in the 5' external transcribed spacer region and of precursor tRNAsec have been described. Here, we used a previously validated semi-nested reverse transcription-polymerase chain reaction strategy to investigate internal transcribed spacer (ITS) I acceptor sites in total RNA from Leishmania (Leishmania) amazonensis. Two distinct spliced leader-containing RNAs were detected indicating that trans-splicing reactions occur at two AG acceptor sites mapped in this ITS region. These data provide further evidence of the wide spectrum of RNA molecules that act as trans-splicing acceptors in trypanosomatids.

Interaction between Telenomus remus and Trichogramma pretiosum in the management of Spodoptera spp.

Interaction betweeen Telenomus remus and Trichogramma pretiosum in the management of Spodoptera spp. The use of egg parasitoids is a promising strategy for Integrated Pest Management (IPM), but different species of parasitoids have greater or lesser control efficiency, depending on the pest species. Recently, not only Anticarsia gemmatalis and Pseudoplusia includens but also Spodoptera cosmioides and S. eridania have been among the key Lepidoptera larvae attacking soybeans. This study evaluated the combination of Telenomus remus and Trichogramma pretiosum for parasitism of eggs of the Spodoptera complex, for better control efficiency and broader spectrum of action among the key pests of soybeans. The experiment was carried out under controlled environmental conditions (25 ± 2ºC; 70 ± 10% RH; and 14 h photophase) in a completely randomized experimental design with seven treatments and 10 replicates with S. frugiperda, S. cosmioides and S. eridania eggs. Each replicate consisted of one egg mass of each Spodoptera species, with approximately 100 eggs offered to the parasitoids. The treatments were: 1) 10 females of T. pretiosum; 2) nine females of T. pretiosum and one female of T. remus; 3) eight females of T. pretiosum and two females of T. remus; 4) seven females of T. pretiosum and three females of T. remus; 5) six females of T. pretiosum and four females of T. remus; 6) five females of T. pretiosum and five females of T. remus, and 7) 10 females of T. remus. The parameter evaluated was the percentage of parasitized eggs. Results showed that treatments combining both parasitoid species with only 1 T. remus for each 9 T. pretiosum (10%) and only 2 T. remus for each 8 T. pretiosum (20%) were enough to significantly increase the parasitism observed on eggs of S. cosmioides and S. frugiperda, respectively. This association of T. pretiosum and T. remus in different proportions is very promising for biological control in IPM programs because it provides wide spectrum of control.

In vitro selection of bacteria with potential for use as probiotics in marine shrimp culture

The objective of this work was to isolate strains of lactic acid bacteria with probiotic potential from the digestive tract of marine shrimp (Litopenaeus vannamei), and to carry out in vitro selection based on multiple characters. The ideotype (ideal proposed strain) was defined by the highest averages for the traits maximum growth velocity, final count of viable cells, and inhibition halo against nine freshwater and marine pathogens, and by the lowest averages for the traits duplication time and resistance of strains to NaCl (1.5 and 3%), pH (6, 8, and 9), and biliary salts (5%). Mahalanobis distance (D²) was estimated among the evaluated strains, and the best ones were those with the shortest distances to the ideotype. Ten bacterial strains were isolated and biochemically identified as Lactobacillus plantarum (3), L. brevis (3), Weissella confusa (2), Lactococcus lactis (1), and L. delbrueckii (1). Lactobacillus plantarum strains showed a wide spectrum of action and the largest inhibition halos against pathogens, both Gram-positive and negative, high growth rate, and tolerance to all evaluated parameters. In relation to ideotype, L. plantarum showed the lowest Mahalanobis (D²) distance, followed by the strains of W. confusa, L. brevis, L. lactis, and L. delbrueckii. Among the analyzed bacterial strains, those of Lactobacillus plantarum have the greatest potential for use as a probiotic for marine shrimp.

Patulin in food: state-of-the-art and analytical trends

Patulin is a mycotoxin produced by several fungal species of the genera Penicillium and Aspergillus, found on several fruit species and, remarkably, in apples and apple products. Patulin has a broad spectrum of toxicity, including carcinogenicity and teratogenicity in animals. Due to the stability of the molecule, considerable amounts of patulin still remain in apple products after processing. This paper reviews different analytical methods for patulin determination and methods to reduce levels of patulin in apple products as well.

Complexation of enalapril maleate with beta-cyclodextrin: NMR spectroscopic study in solution

A detailed NMR (¹H , COSY, ROESY) spectroscopic study of complexation of enalapril maleate with beta-cyclodextrin was carried out. The ¹H NMR spectrum of enalapril maleate confirmed the existence of cis-trans equilibrium in solution, possibly due to hindered rotation along the amide bond. The cis-trans ratio remained almost the same in the presence of beta-cyclodextrin but in one case it was found significantly different which suggests a catalytic role of beta-cyclodextrin in the isomerization. ¹H NMR titration studies confirmed the formation of an enalapril-beta-cyclodextrin inclusion complex as evidenced by chemical shift variations in the proton resonances of both the host and the guest. The stoichiometry of the complex was determined to be 2:1 (guest: host). The mode of penetration of the guest into the beta-cyclodextrin cavity as well as the structure of the complex were established using ROESY spectroscopy.

Selective and non-extractive spectrophotometric determination of cefdinir in formulations based on donor-acceptor complex formation

Cefdinir has broad spectrum of activity and high prescription rates, hence its counterfeiting seems imminent. We have proposed a simple, fast, selective and non-extractive spectrophotometric method for the content assay of cefdinir in formulations. The method is based on complexation of cefdinir and Fe under reducing condition in a buffered medium (pH 11) to form a magenta colored donor-acceptor complex (λ max = 550 nm; apparent molar absorptivity = 3720 L mol-1 cm-1). No other cephalosporins, penicillins and common excipients interfere under the test conditions. The Beer's law is followed in the concentration range 8-160 µg mL-1.

Resistance spectra of wheat cultivars and virulence diversity of Magnaporthe grisea isolates in Brazil

Seventy-two monoconidial isolates of Magnaporthe grisea were obtained from the States of Mato Grosso do Sul and Paraná. The isolates were inoculated on seedlings of 20 wheat (Triticum aestivum) cultivars under greenhouse conditions. The virulence diversity of M. grisea was assessed based on compatible and incompatible reactions of leaf blast on wheat cultivars. Fifty-four distinct virulence patterns were identified on test cultivars among the isolates collected from the two wheat growing States. Sixteen of these isolates corresponding to 22.2% showed similar virulence pattern. None of the wheat cultivars was resistant to all isolates of M. grisea, but the cultivars differed in degree of resistance as measured by the relative spectrum of resistance (RSR) and disease index (DI). Among the cultivars the RSR ranged from 0 to 53.3% and DI from 0.4662 to 0.9662 (0 to 1 scale). The wheat cultivar BR18 exhibited a broad resistance spectrum in relation to the rest of the tested cultivars to the isolates of M. grisea, and can be used in wheat resistance breeding.

Synthesis and characterization of solid 2-methoxycinnamylidenepyruvic acid

The 2-methoxycinnamylidenepyruvic acid (2-MeO-HCP) was synthesized and characterized for nuclear magnetic resonance (¹H and 13C NMR), mass spectrometry (MS), Infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The application of DSC for purity determination is well documented in literature and is used in the analysis of pure organic compounds. The molecular geometry and vibrational frequencies of 2-MeO-HCP have been calculated.

Application Volumes and Sizes of Droplets for the Application of Diquat Herbicide in the Control of Eichhornia crassipes

The objective of this study was to evaluate the efficiency of application and the efficacy of control of water hyacinth (Eichhornia crassipes) with the use of the diquat herbicide done with two application volumes associated to three droplets classes. Three experiments were conducted; in the first, the application efficiency was evaluated; in the second, the control efficiency and; in the third, the droplet spectrum. They were conducted in a completely randomized design with five, four and six replicates, respectively. The treatments were arranged in a 2 x 3 factorial design, with two application volumes (100 and 200 L ha-1) and three droplets classes (fine, extremely coarse and ultra-coarse) to the first and second experiment and the third comprised two diquat concentrations in spray solution (2 and 4 g i.a. L-1) and three droplets classes (fine, extremely coarse and ultra-coarse). The application efficiency was determined by the coverage by droplets, spray deposition and active ingredient of the herbicide (diquat). The efficacy was measured by the control and the percentage of plants with regrowth at 50 days after application. The spectrum of droplets produced per each nozzle model used to obtain the droplets classes were analyzed. According to the parameters analyzed, using the droplets classes extremely thick and ultra thick can provide greater certainty in the application of diquat in the aquatic environment associated with the deposition of the active and sufficient coverage to control Eichhornia crassipes with both application volumes

Reproductive biology of the herkogamous vine Chiococca alba (L.) Hitchc. (Rubiaceae) in the Atlantic Rain Forest, SE Brazil

The family Rubiaceae comprises a wide spectrum of floral mechanisms and two of them seem to be common in certain groups, e.g., distyly in Rubioidae and styllar pollen in Ixoroidae. These mechanisms include herkogamy, which is interpreted as a strategy that avoids self-pollination. This is the first report on the reproductive biology of Chiococca alba, a species that is widely distributed in America. We studied floral biology and the mating system, which were evaluated through fruit set comparisons after controlled crosses (self- and cross-pollinations and test for apomixis), as well as through the evaluation of pollen tube growth resulting from these controlled crosses. Flowers of C. alba are herkogamous, cream, protandrous and lasted for two days. No measurable nectar was found, despite the presence of a nectary-like structure at the base of the corolla tube. Chiococca alba is a preferentially self-incompatible species, but self-pollination and apomixis also contribute to the natural fruit-set. Its reproductive strategy (herkogamy associated with protandry) is different from that expected for members of Chiococceae tribe (i.e., styllar pollen presentation).

A comparative analysis of preprocessing techniques of cardiac event series for the study of heart rhythm variability using simulated signals

In the present study, using noise-free simulated signals, we performed a comparative examination of several preprocessing techniques that are used to transform the cardiac event series in a regularly sampled time series, appropriate for spectral analysis of heart rhythm variability (HRV). First, a group of noise-free simulated point event series, which represents a time series of heartbeats, was generated by an integral pulse frequency modulation model. In order to evaluate the performance of the preprocessing methods, the differences between the spectra of the preprocessed simulated signals and the true spectrum (spectrum of the model input modulating signals) were surveyed by visual analysis and by contrasting merit indices. It is desired that estimated spectra match the true spectrum as close as possible, showing a minimum of harmonic components and other artifacts. The merit indices proposed to quantify these mismatches were the leakage rate, defined as a measure of leakage components (located outside some narrow windows centered at frequencies of model input modulating signals) with respect to the whole spectral components, and the numbers of leakage components with amplitudes greater than 1%, 5% and 10% of the total spectral components. Our data, obtained from a noise-free simulation, indicate that the utilization of heart rate values instead of heart period values in the derivation of signals representative of heart rhythm results in more accurate spectra. Furthermore, our data support the efficiency of the widely used preprocessing technique based on the convolution of inverse interval function values with a rectangular window, and suggest the preprocessing technique based on a cubic polynomial interpolation of inverse interval function values and succeeding spectral analysis as another efficient and fast method for the analysis of HRV signals

Cytokines produced by susceptible and resistant mice in the course of Paracoccidioides brasiliensis infection

Paracoccidioidomycosis (PCM) is the most prevalent deep mycosis in Latin America and presents a wide spectrum of clinical manifestations. We established a genetically controlled murine model of PCM, where A/Sn mice develop an infection which mimics the benign disease (immune responses which favor cellular immunity) and B10.A animals present the progressive disseminated form of PCM (preferential activation of B cells and impairment of cellular immune responses). To understand the immunoregulatory phenomena associated with resistance and susceptibility in experimental PCM, A/Sn and B10.A mice were studied regarding antigen-elicited secretion of monokines (TNF-a and TGF-ß) and type-1 (IL-2 and IFN-g) and type-2 (IL-4,5,10) cytokines. Total lymph node cells from resistant mice infected ip with P. brasiliensis produced early and sustained levels of IFN-g and IL-2; type-2 cytokines (IL-4 and IL-5) started to appear 8 weeks after infection. In contrast, susceptible mice produced low levels of IFN-g concomitant with significant levels of IL-5 and IL-10 early in the infection. In the chronic phase of the disease, susceptible animals presented a transitory secretion of IL-2, and IL-4. In the pulmonary infection IL-4, IL-5 and IL-10 were preferentially detected in the lung cells washings of susceptible animals. After in vitro challenge with fungal antigens, normal peritoneal macrophages from B10.A mice secreted high levels of TGF-ß and low levels of TNF-a. In contrast, macrophages from A/Sn animals released high levels of TNF-a associated with a small production of TGF-ß. The in vivo depletion of IFN-g not only abrogated the resistance of A/Sn mice but also diminished the relative resistance of B10.A animals. The in vivo depletion of IL-4 did not alter the disease outcome, whereas administration of rIL-12 significantly enhanced resistance in susceptible animals. Taken together, these results suggest that an early secretion of high levels of TNF-a and IFN-g followed by a sustained secretion of IL-2 and IFN-g plays a dominant role in the resistance mechanisms to P. brasiliensis infection. In contrast, an early and ephemeral secretion of low levels of TNF-a and IFN-g associated with production of IL-5, IL-10 and TGF-ß characterizes the progressive disease of susceptible animals.

Amifostine (WR-2721), a cytoprotective agent during high-dose cyclophosphamide treatment of non-Hodgkin's lymphomas: a phase II study

Clinical trials indicate that amifostine may confer protection on various normal tissues without attenuating anti-tumor response. When administered prior to chemotherapy or radiotherapy, it may provide a broad spectrum of cytoprotection including against alkylating drugs. The mechanism of protection resides in the metabolism at normal tissue site by membrane-bound alkaline phosphatase. Toxicity of this drug is moderate with hypotension, nausea and vomiting, and hypocalcemia being observed. We report a phase II study using amifostine as a protective drug against high-dose cyclophosphamide (HDCY) (7 g/m2), used to mobilize peripheral blood progenitor cells (PBPC) and to reduce tumor burden. We enrolled 29 patients, 22 (75.9%) affected by aggressive and 7 (24.1%) by indolent non-Hodgkin's lymphoma (NHL), who were submitted to 58 infusions of amifostine and compared them with a historical group (33 patients) affected by aggressive NHL and treated with VACOP-B followed by HDCY. The most important results in favor of amifostine were the reduction of intensity of cardiac, pulmonary and hepatic toxicity, and a significant reduction of frequency and severity of mucositis (P = 0.04). None of the 29 patients died in the protected group, while in the historical group 2/33 patients died because of cardiac or pulmonary toxicity and 2 patients stopped therapy due to toxicity. Amifostine did not prevent the aplastic phase following HDCY. PBPC collection and hematological recovery were adequate in both groups. The number of CFU-GM (colony-forming units-granulocyte/macrophage) colonies and mononuclear cells in the apheresis products was significantly higher in the amifostine group (P = 0.02 and 0.01, respectively). Side effects were mild and easily controlled. We conclude that amifostine protection should be useful in HDCY to protect normal tissues, with acceptable side effects.

The effect of porphyrins on normal and transformed mouse cell lines in the presence of visible light

Photodynamic therapy consists of the uptake of a photosensitizing dye, often a porphyrin, by tumor tissue and subsequent irradiation of the tumor with visible light of an appropriate wavelength matched to the absorption spectrum of the photosensitizing dye. This class of molecules produces reactive oxygen species when activated by light, resulting in a direct or indirect cytotoxic effect on the target cells. Photodynamic therapy has been used in the treatment of cancer but the technology has a potential for the treatment of several disease conditions mainly because of its selectivity. However, it is not clear why the porphyrins are retained preferentially by abnormal tissue. This paper describes a study of the effect of the association of porphyrin and visible light on two mouse fibroblast cell lines: A31, normal cells and B61, an EJ-ras transformed variant of A31. Two water-soluble porphyrins were used, a positively charged one, tetra(N-methyl-4-pyridyl)porphyrin chloride, and a negatively charged one, tetra(4-sulfonatophenyl)porphyrin-Na salt (TPPS4) in order to assess the effect on cell survival. The results suggest that the B61 cell line is more sensitive to incubation with the anionic porphyrin (TPPS4) followed by light irradiation and that the anionic porphyrin is more efficient in killing the cells than the cationic porphyrin.