Effect of a single dose of inducers and inhibitors on the rate of synthesis of cytochromes and reductases in liver organelles

Summary Measurement of the effect of drugs on the in vivo rates of synthesis of rabbit liver organelle bound proteins were measured following individual treatments with the inducers phenobarbital, 3-methylcholanthrene and PCB (a mixture of polychlorinated biphenyls) and the inhibitors, cycloheximide, aflatoxin 13, chloramphenicol and actinomycin D. Following their isolation from a homogenate containing the combined livers of ¹⁴C-leucine injected experimental animals and 3H-leucine injected control animals, purified fractions of the following proteins were prepared: microsomal cytochrome b₅, cytochrome P-450, NADH-cytochrome b₅ reductase, NADPH-cytochrome P-450 reductase and proteolipids, outer mitochrondrial membrane cytochrome b5, NADH-cytochrome b₅ reductase and proteolipids, inner mitochrondrial membrane cytochrome c, NADH dehydrogenase and proteolipids, intermitochrondrial membrane cytochrome b₅ and circulating serum albumin.This research was supported by grants from the PSC-BHE Research Award Program of The City University of New York, US Public Health Service Grant SO 7 PRO7132-07 and the Alma Toorock Fund for Cancer Research.     

Illustrated and Annotated Checklist of Brazilian Gall Morphotypes

The analysis on nine inventories on the richness and diversity of galling herbivores in Brazil accounted for 806 gall systems occurring in 443 host-plant species from 74 plant families. This checklist of the Brazilian gall morphotypes proposes seven standardized morphotypes and five additional shapes that group the majority of the three-dimensional shapes reported in literature. Criteria are proposed to standardize the terminology, and a critical analysis is provided aiming to avoid possible inconsistencies in order to generate easily comparable data in future inventories. The morphotypes are herein catalogued in alphabetical order, accompanied by a conceptual definition, an illustration, and examples that best represent the shape. It is proposed that the inventories should present at least the (1) host-plant species, (2) galling herbivore species or its identification to the lowest possible taxonomic level, (3) host-plant galled organ and gall position, (4) gall morphotype, (5) gall color and registration of indumentum when present, (6) gall phenological and developmental data, (7) association with other trophic levels, and (8) additional information, such as dimension, and number of chamber(s).     

Antimicrobial activity of fractions and compounds from Calophyllumbrasiliense (Clusiaceae/Guttiferae)

Calophyllum brasiliense (Clusiaceae/Guttiferae) is a native Brazilian medicinal plant traditionally used against several diseases, including infectious pathologies. Crude methanolic extracts (CME) and two fractions, denoted non-polar (soluble in chloroform) and polar (nonsoluble in chloroform), were prepared from different parts of the plant (roots, stems, leaves, flowers and fruits) and studied. The following compounds were isolated and tested against pathogenic bacteria and yeasts by determination of the minimal inhibitory concentration (MIC): brasiliensic acid (1), gallic acid (2), epicatechin (3), protocatechuic acid (4), friedelin (5) and 1,5-dihydroxyxanthone (6). The results indicated that all the parts of the plant exhibited antimicrobial activity against Gram-positive bacteria, which are selectively inhibited by components of C. brasiliense. No activity was observed against Gram-negative bacteria and yeasts tested. Regarding the isolated compounds, substance 4 showed antimicrobial activity against all the tested microorganisms, whereas compound 6 exhibited antimicrobial activity only against Gram-positive bacteria. The results from the current study confirm and justify the popular use of this plant to treat infectious processes.     

Overexpression of a synthetic gene encoding human alpha interferon in Escherichia coli

Interferons (IFNs) represent an important defense mechanism in vertebrates. In this work, we describe gene synthesis and assembly using the polymerase chain reaction as a method for single-step synthesis of DNA sequences. The oligonucleotides designed were based on Escherichia coli codon usage and two genes of IFN were synthesized: one containing a DNA sequence already known and the other, a mutated form in which two cysteine amino acid residues were replaced by serines in an attempt to improve the stability of the protein. DNA sequences were cloned into pAE, an E. coli vector that allows heterologous protein expression with or without a histidine tag. Recombinant human interferons (rhIFNs) were identified by Western blotting and ELISA using anti-human interferon polyclonal antibodies. Purification of the recombinant His-tagged proteins was achieved in a single step by Ni(2+)-charged column chromatography while proteins without His-tag were purified by extensively washing the inclusion bodies, the final yields being approximately 210 and 75mg/L, respectively. The rhIFNs expressed within this system were biologically active ( approximately 1,1x10(8)IU/mg) based on antiviral assay. The combined methodologies described here proved to be cost-effective and could be extended to other genes/proteins of interest.     

Anatomy and photosynthetic parameters of roots and leaves of two shade-adapted orchids, Dichaea cogniauxiana Shltr. and Epidendrum secundum Jacq.

This study compares photosynthetic and structural features of Dichaea cogniauxiana and Epidendrum secundum leaves and roots. The diurnal titratable acidity fluctuations indicated crassulacean acid metabolism (CAM) in E. secundum leaves, associated with anatomical features like thick cuticle, large and vacuolated cells, and reduced stomata size and frequency. Roots of both species had chloroplasts in their cortical parenchyma. However, neither the roots nor D. cogniauxiana leaves did show tissue sap acidity fluctuations. This indicates C₃ metabolism in these organs. This lack of oscillation of organic acids in Epidendrum roots was at odds with a CAM-like ¹³C ratio, suggesting that in spite of active CO₂ fixation in roots during the day, the bulk of carbon is imported from the leaves. Roots of both species showed Fv/Fm, ΔF/Fm′, ETR values similar to reports from other non-foliar photosynthetic organs. Besides reducing root carbon cost, root photosynthesis may also be important by alleviating potential hypoxia, since water-saturated velamen severely impedes the gas exchange between radicular cortex.     

Secondary stem anatomy and uses of four drought-deciduous species of a tropical dry forest in México

Wood and bark anatomy and histochemistry of Acacia bilimekii Humb. & Bonpl., Acacia cochliacantha Mcbride, Conzatia nultiflora (Rob) Stand. and Guazuma ulmifolia Lam. are described from stem samples collected in a tropical dry forest (Morelos, Mexico). Enzyme activities were tested in tangential, radial and transverse cuts of fresh material. Histochemistry and stem anatomy were studied on similar cuts previously softened in a solution of water-glicerol-PEG. Our results show that the anatomical patterns of bark and wood, as well as the histochemical patterns and specific gravity, are influenced by water accessibility and climate; these patterns could guarantee mechanical and anti-infection strategies to support extreme conditions. Enzyme cytochemistry reveals biochemical activities probably related to lipid utilization routes for the lignification processes and for synthesis of extractives; these results suggest that the formation and maturation of woody tissue is very active at the beginning of the rainy season. These species are widely used by the local population. Traditional uses include firewood, dead and live fences, fodder, construction, supporting stakes, handcrafts, farming tools, extraction of tanning products, and medicine. There is no relationship between use and abundance. Alternative uses are proposed according to a density index.     

Aluminium localization and toxicity symptoms related to root growth inhibition in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) seedlings

We correlated root growth inhibition with aluminium (Al³⁺) localization and toxicity symptoms in rice roots using seedlings of two genotypes (tolerant and sensitive) that were exposed to different AlCl₃ concentrations. Al³⁺ localization was evaluated by hematoxylin in primary roots and by morin in cross-sections of the root tips. Neutral invertase enzyme activity and callose (1→3, β-d-glucan) accumulation were observed and compared with Al³⁺ accumulation sites. Root growth was inhibited by Al³⁺ in a concentration-specific manner and proportional to the increase of hematoxylin staining, being more pronounced in the sensitive genotype. Morin staining showed the presence of Al³⁺ deep within the roots of the sensitive genotype, indicating that the metal was able to penetrate beyond the first few cell layers. In the tolerant genotype, Al³⁺ penetration was restricted to the first two cell layers. Ruptures in exodermis and epidermis layers by lateral root protrusions in both genotypes allowed Al³⁺ to enter into the roots. More intense activity of invertase in roots of the tolerant genotype was also observed, which could be related to greater root growth of this cultivar when submitted to Al³⁺ stress. Moreover, Al³⁺-induced callose accumulation was a late response occurring in the same areas where Al³⁺ was present.     

Evaluation of Oxidative Stress in Overweight Subjects With or Without Metabolic Syndrome

Although oxidative stress is considered the underlying mechanism by which dysfunctional metabolism occurs in obese subjects, there are few studies on oxidative stress in overweight subjects. The objective of this study was to verify the influence of metabolic syndrome (MetS) on oxidative stress and antioxidant defense in overweight subjects. There were 123 subjects (50 in the control group and 73 in the overweight group) chosen to participate in this cross‐sectional study. The control group included 50 healthy individuals with a BMI between 20 and 24.9 kg/m² and without MetS. The overweight group included 73 subjects with a BMI between 25 and 29.9 kg/m². Overweight subjects were divided into two groups: with MetS (29 subjects) and without MetS (44 subjects). Control group and overweight group subjects without MetS showed no differences in oxidative stress parameters and total antioxidant capacity (TRAP). Overweight subjects with MetS had higher hydroperoxide concentrations measured by chemiluminescence compared to the control group (P < 0.05), higher hydroperoxide and hydrogen peroxide concentrations determined by ferrous oxidation‐xylenol orange assay compared to overweight subjects without MetS (P < 0.001), and higher advanced oxidation protein product (AOPP) concentrations (P < 0.001) compared to the other groups. AOPP was directly correlated with uric acid concentrations. Overweight subjects with MetS had lower TRAP concentrations compared to the control group (P < 0.001). In conclusion, this study showed that overweight subjects with MetS, in contrast to overweight subjects without MetS, have a redox imbalance characterized by increased plasma oxidation and reduced antioxidant capacity.     

Purification and characterization of aprotinin from porcine lungs

Aprotinin, the most studied serine proteinase inhibitor, was isolated from porcine lung for the first time. The purified porcine aprotinin had an Mr value of ∼7 kDa. It cross-reacted with polyclonal serum anti-commercial aprotinin. About 1 μg porcine aprotinin inhibited 6 μg trypsin whereas 1 μg commercial soybean inhibitor inhibited only 1 μg trypsin. The aprotinin gene was also isolated from porcine lung: the deduced amino acid sequence showed 74% identity to bovine aprotinin.     

Unravelling the structure and function of the petal appendages in the tribe Schwenckieae (Solanaceae)

• The tribe Schwenckieae (Solanaceae) is characterised by the presence of appendages on the corolla, a diagnostic trait for the group. These appendages constitute a median distal projection of the three‐lobed petal and occur in the genera Melananthus and Schwenckia but are absent in Heteranthia.
• We investigated the micromorphology and anatomical structure of the appendages and lateral petal lobes of Schwenckia americana (two varieties), S. angustifolia, S. curviflora and S. novaveneciana, and Melananthus fasciculatus. We also performed histochemical tests to determine if the appendages are involved in the production of volatiles, acting as a fragrance secretory structure (osmophore).
• The appendages have a uniseriate epidermis, whose cells store phenolics and lipids. The parenchyma is starch‐rich just prior to anthesis in all species studied. The sensory test and anatomical analyses identified scent‐secreting tissues, not only in the appendages, but also in the lateral petal lobes, whose cells are papillose with a sculptured surface. The α‐naphthol p‐phenylenediamine (NADI) reaction detected volatile (essential oils) compounds in S. americana var. americana and S. americana var. angustifolia.
• We demonstrated the secretory tissues and the production of lipids in the corolla appendages of Schwenckia and Melananthus, which indicate their osmogenic function and probable scent emission to attract pollinators.