Metabolic profiling by ultra-performance liquid chromatography-mass spectrometry and parallel factor analysis for the determination of disease biomarkers in Eucalyptus

In this article, we present and discuss an alternative for data analysis of the metabolic profiles of both healthy Eucalyptus globulus and those infected with the Mycosphaerella leaf disease. The crude extracts were analyzed by reversed-phase ultra performance liquid chromatography-mass spectrometry. In order to glean the most useful information from these complex measurements, parallel factor analysis (PARAFAC) was employed for pattern recognition. After PARAFAC modeling, inspection of the scores and loadings graph allowed distinction of the healthy from the infected E. globulus samples and determination of biomarkers related to the biotic stress. The assessment of the monoisotopic masses and the fragmentation patterns allowed the identification of these biomarkers. It is hoped that the proposed method can be used for the diagnosis of diseases in plants, as well as to provide additional insight into the plant’s defense mechanism. Potentially, this may demonstrate the advantages of employing high order chemometric techniques in metabolomic data analysis.     

MHC Class I Chain-Related Gene A Polymorphisms and Linkage Disequilibrium with HLA-B and HLA-C Alleles in Ocular Toxoplasmosis

This study investigated whether polymorphisms of the MICA (major histocompatibility complex class I chain-related gene A) gene are associated with eye lesions due to Toxoplasma gondii infection in a group of immunocompetent patients from southeastern Brazil. The study enrolled 297 patients with serological diagnosis of toxoplasmosis. Participants were classified into two distinct groups after conducting fundoscopic exams according to the presence (n = 148) or absence (n = 149) of ocular scars/lesions due to toxoplasmosis. The group of patients with scars/lesions was further subdivided into two groups according to the type of the ocular manifestation observed: primary (n = 120) or recurrent (n = 28). Genotyping of the MICA and HLA alleles was performed by the polymerase chain reaction-sequence specific oligonucleotide technique (PCR-SSO; One Lambda®) and the MICA-129 polymorphism (rs1051792) was identified by nested polymerase chain reaction (PCR-RFLP). Significant associations involving MICA polymorphisms were not found. Although the MICA*002~HLA-B*35 haplotype was associated with increased risk of developing ocular toxoplasmosis (P-value = 0.04; OR = 2.20; 95% CI = 1.05–4.60), and the MICA*008~HLA-C*07 haplotype was associated with protection against the development of manifestations of ocular toxoplasmosis (P-value = 0.009; OR: 0.44; 95% CI: 0.22–0.76), these associations were not statistically significant after adjusting for multiple comparisons. MICA polymorphisms do not appear to influence the development of ocular lesions in patients diagnosed with toxoplasmosis in this study population.     

Real-time PCR method to quantify Sp245 strain of Azospirillum baldaniorum on Brachiaria grasses under field conditions

Plant and Soil - Soil microbiome roles in agriculture is becoming more and more important. This importance is also reflected on the way plants are seen: complex organisms formed by the plant itself...     

Temperature dependence of the germination of tomato seeds

1. 1.|The germination of tomato “C38” seeds exposed to periodical white incandescent light occurs from 6.0° ± 0.2°C to 37.5° ± 0.2°C, being rate-limited for 10.3° T 25.9°C, and elsewhere limited by the germination capacity.

2. 2.|Rate averages are linearly T-dependent outside their optimum range (25.9° T 29.5°C) and rate variances are typically heterogeneous.

3. 3.|The smooth curvilinear Arrhenius plot indicates that diffusion processes cannot be rate-limiting outside the interval 25.9° T 29.5°C, whereas phase transitions and (or) transconformation of proteins may limit the rate above 34.9°C and, by opposite effects, below 15.3°C.

4. 4.|The thermal communication between the environment and the germinating seed proceeds by a temperature signal which is quenched by random thermal noise at T 11.2°C and at T 34.0°C.

TRANSPARENT EXOPOLYMER PARTICLES FORMATION FROM CAPSULES OF ANABAENA SPIROIDES (CYANOBACTERIA) IN CULTURE1

We report the production of large numbers of transparent exopolymer particles (TEP) from polysaccharidic capsules of Anabaena spiroides Kleb. in cultures. Two biotic pathways of TEP formation were observed: (1) fragmentation of small portions of the capsules, which occurred throughout the growth phases; and (2) transformation of the whole polysaccharidic capsules into TEP, following cellular lyses in the aging culture. Photographic documentation of these processes was performed after staining small aliquots of the samples with Alcian Blue and negative staining with India ink. Concentrations of TEP were determined in distinct culture growth phases using semiquantitative Alcian Blue staining. Concentrations of TEP increased throughout the experimental time, while Alcian Blue remaining in solution decreased. Decreasing concentrations of chl a indicated cellular death, and by the end of the experiment, TEP formed by both pathways accumulate in the culture medium. These results show that virtually all dead chains of A. spiroides are transformed into TEP in the aged culture.     

Non‐continuous reproductive phenology of animal‐dispersed species in young forest restoration plantings

Tree species that produce resources for fauna are recommended for forest restoration plantings to attract pollinators and seed dispersers; however, information regarding the flowering and fruiting of these species during early growth stages is scarce. We evaluated the reproductive phenology of animal‐dispersed tree species widely used in Atlantic Forest restoration. We marked 16 animal‐dispersed tree species in 3‐ to 8‐year‐old forest restoration plantings in Itu‐São Paulo, southeast Brazil. We noted the age of the first reproductive event, flowering and fruiting seasonality, percentage of trees that reached reproductive stages, and intensity of bud, flower, and fruit production for each species. Flowering and fruiting are seasonal for most species; only two, Cecropia pachystachya and Ficus guaranitica, exhibited continuous flowering and fruiting throughout the year; we also identified Schinus terebinthifolia and Dendropanax cuneatus fruiting in the dry season during resource scarcity. Therefore, we recommend all as framework species, that is, species that are animal‐dispersed with early flowering and fruiting potential, for forest restoration. Further, we recommend identifying and planting similar animal‐dispersed tree species that produce fruits constantly or in the dry season to maximize fauna resource availability throughout the year in tropical forest restoration plantings. Abstract in Portuguese is available with online material     

Production of biosurfactant by a genetically-modified strain ofBacillus subtilis expressing green fluorescent protein

Biosurfactant production was investigated using two strains ofBacillus subtilis, being one a reference strain (B. subtilis 1012) and the other a genetically-modified strain (B. subtilis W1012) made able to produce the green fluorescent protein (GFP). A new method based on oil displacement technique was set up to measure the biosurfactant level in the medium. Although the tested microorganisms showed similar results in terms of cell growth parameters, the recombinant strain, besides expressing GFP, exhibited an average yield of extracellular surfactant on biomass (Y _{B/X, av} =239 mg_B g_x ^?1) more than twice that of the reference strain. The ability of the genetically-modified strain to simultaneously overproduce biosurfactant and GFP even at low cell concentration makes it an interesting candidate for possible use as a biological index-finger to monitor cell viability in bioremediation and oil recovery operations.     

Cardiorespiratory reflexes and aquatic surface respiration in the neotropical fish tambaqui (<Emphasis Type="Italic">Colossoma macropomum</Emphasis>): acute responses to hypercarbia

We examined the cardiorespiratory responses to 6 h of acute hypercarbia (1, 2.5, and 5% CO₂) in intact and gill-denervated (bilateral denervation of branchial branches of cranial nerves IX and X) tambaqui, Colossoma macropomum. Intact fish exposed to 1 and 2.5% CO₂ increased respiratory frequency (f_R) and ventilation amplitude (V_AMP) slowly over a 1- to 3-h period. Denervated fish did not show this response, suggesting that tambaqui possess receptors in the gills that will produce excitatory responses to low levels of hypercarbia (1 and 2.5% CO₂) if the exposure is prolonged. The cardiac response to stimulation of these receptors with this level of CO₂ was a tachycardia and not a bradycardia. During exposure to 5% CO₂, intact fish increased f_R and V_AMP, and showed a pronounced bradycardia after 1 h. After 2 h, the heart rate (f_H) started to increase, but returned to control values after 6 h. In denervated fish, the increase in f_R was abolished. The slow increase in V_AMP and the bradycardia were not abolished, suggesting that these changes arose from extra-branchial receptors. Neither intact nor denervated fish developed the swelling of the lower lip or performed aquatic surface respiration, even after 6 h, suggesting that these are unique responses to hypoxia and not hypercarbia.Abbreviations ASR aquatic surface respiration - f_H heart frequency - f_R respiratory frequency - V_AMP ventilation amplitude - _TOT total ventilation