The effects of the C-terminal amidation of mastoparans on their biological actions and interactions with membrane-mimetic systems

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structure-activity relationship of mastoparan analogs: effects of the number and positioning of Lys residues on secondary structure, interaction with membrane-mimetic systems and biological activity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of elevated atmospheric carbon dioxide on the biological control of coffee leaf rust under controlled conditions

The effect of elevated atmospheric CO2 concentration on biological control of coffee leaf rust, caused by Hemileia vastatrix, was evaluated by leaf disc assay, under controlled conditions. The biocontrol agents Bacillus subtilis, Bacillus pumilus and Lecanicillium longisporum were applied 24h before, 24h after, and simultaneously with the H. vastatrix on leaf discs (diameter of 1.5cm). The CO2 concentrations tested were: 380, 430, 700 and 1300ppm for B. subtilis and B. pumilus; and 380, 430, 670 and 1200ppm for L. longisporum. The antagonists were not affected by CO2 concentrations. B. subtilis was the most effective in controlling the disease when applied before and simultaneously with pathogen.

Biological and structural characterization of a new PLA(2) from the Crotalus durissus collilineatus venom

In the present article we report on the biological characterization and amino acid sequence of a new basic Phospholipases A(2) (PLA(2)) isolated from the Crotalus durissus collilineatus venom (Cdcolli F6), which showed the presence of 122 amino acid residues with a pI value of 8.3, molecular mass of 14 kDa and revealed an amino acid sequence identity of 80% with crotalic PLA(2)s such as Mojave B, Cdt F15, and CROATOX. This homology, however, dropped to 50% if compared to other sources of PLA(2)s such as from the Bothrops snake venom. Also, this PLA(2) induced myonecrosis, although this effect was lower than that of BthTx-I or whole crotoxin and it was able to induce a strong blockage effect on the chick biventer neuromuscular preparation, independently of the presence of the acid subunid (crotapotin). The neurotoxic effect was strongly reduced by pre-incubation with heparin or with anhydrous acetic acid and rho-BPB showed a similar reduction. The rho-BPB did not reduce significantly the myotoxic activity induced by the PLA(2), but the anhydrous acetic acid treatment and the pre-incu-bation of PLA(2) with heparin reduced significantly its effects. This protein showed a strong antimicrobial activity against Xanthomonas axonopodis passiflorae (Gram-negative), which was drastically reduced by incubation of this PLA(2) with rho-BPB, but this effect was marginally reduced after treatment with anhydrous acetic acid. Our findings here allow to speculate that basic amino acid residues on the C-terminal and molecular regions near catalytic site regions such as Calcium binding loop or rho-wing region may be involved in the binding of this PLA(2) to the molecular receptor to induce the neurotoxic effect. The bactericidal effect, however, was completely dependent on the enzymatic activity of this protein.

Biological indicators of stress in pacu (Piaractus mesopotamicus) after capture

Os efeitos da captura (perseguição, contenção em puçá e exposição aérea) no perfil sanguíneo do cortisol, glicose, cloreto, sódio, potássio, cálcio e na osmolaridade, hematócrito, hemoglobina, número de células vermelhas (CV) e volume corpuscular médio (VCM) foram investigados no pacu (Piaractus mesopotamicus). Um total de 132 peixes (49,7 ± 11,7 g) foi submetido à captura com 3 ou 5 minutos de exposição aérea. Nove peixes de cada tratamento foram amostrados 5, 15, 30, 60 minutos e 24 horas depois e outros nove peixes foram amostrados antes da captura e considerados controle. A captura resultou em aumento do cortisol e glicose no sangue 30 e 5 minutos depois da captura, respectivamente, independente do tempo de exposição aérea. Ambos os indicadores recuperaram os valores controle em 24 horas. Nos dois grupos de peixes, o cloreto plasmático diminuiu 60 minutos após captura e não recuperou os valores controle, enquanto o sódio sérico aumentou entre 15 e 30 minutos recuperando a condição controle em 24 horas. Não houve alteração significativa nos valores de potássio, cálcio, osmolaridade ou no hematócrito, hemoglobina, CV e VCM como consequência da captura. Os estressores sequenciais aplicados no pacu durante a captura ativaram o eixo cérebro-pituitária-interrenal (respostas do cortisol e glicose), mas a ativação do eixo cérebro-sistema simpático-células cromafins foi aparentemente moderada (respostas iônicas e hematológicas).

Effects of elevated calcium on motor and exploratory activities of rats

The effects of serum and brain calcium concentration on rat behavior were tested by maintaining animals on either distilled water (N = 60) or water containing 1% calcium gluconate (N = 60) for 3 days. Animals that were maintained on high calcium drinking water presented increased serum calcium levels (control = 10.12 ± 0.46 vs calcium treated = 11.62 ± 0.51 µg/dl). Increase of brain calcium levels was not statistically significant. In the behavioral experiments each rat was used for only one test. Rats that were maintained on high calcium drinking water showed increased open-field behavior of ambulation (20.68%) and rearing (64.57%). on the hole-board, calcium-supplemented animals showed increased head-dip (67%) and head-dipping (126%), suggesting increased ambulatory and exploratory behavior. The time of social interaction was normal in animals maintained on drinking water containing added calcium. Rats supplemented with calcium and submitted to elevated plus-maze tests showed a normal status of anxiety and elevated locomotor activity. We conclude that elevated levels of calcium enhance motor and exploratory behavior of rats without inducing other behavioral alterations. These data suggest the need for a more detailed analysis of several current proposals for the use of calcium therapy in humans, for example in altered blood pressure states, bone mineral metabolism disorders in the elderly, hypocalcemic states, and athletic activities.

Effects of different temperatures on the life history of Evania appendigaster L. (Hymenoptera: Evaniidae), a solitary oothecal parasitoid of Periplaneta americana L. (Dictyoptera: Blattidae)

The influence of temperatures on the life parameters of the solitary oothecal parasitoid Evania appendigaster, was investigated in the laboratory. Parasitized oothecae of Periplaneta americana were left to develop under seven constant temperatures: 15, 17, 20, 25, 30, 35, and 40 degrees C. At the end, we found that: (i) E. appendigaster was able to complete development within the temperature range of 17-34 degrees C; (ii) mean adult longevity decreased as temperature increased, with the temperature of 40 degrees C being fatal in a matter of hours; (iii) males lived longer than females between 15 and 30 degrees C; (iv) adult emergence rate was the highest at 25 degrees C, and (v) no wasps emerged at 15 or 40 degrees C. Non-emerged oothecae contained either unhatched eggs or dead larvae. We determined the theoretical lower developmental threshold and thermal constant for the complete development as 12.9 degrees C and 584.8 day-degrees for males, and 13.1 degrees C and 588.2 day-degrees for females, respectively. A good balance between faster development, maximum adult longevity and good egg viability was obtained between 25-30 degrees C, and that would be the best temperature range for rearing E. appendigaster. (C) 2009 Elsevier B.V. All rights reserved.

Bovine Biofertilizer and Water Regime Effects on Growth and Bulb Quality of Garlic

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mitochondrial genotype segregation and effects during mammalian development: Applications to biotechnology

Mitochondria are endosymbiotic organelles responsible for energy production in practically every eukaryotic cell. Their uniparental fashion of inheritance, maternally inherited in mammals, and the homogeneity of mitochondrial DNA (mtDNA) within individuals and matrilineages, are biological phenomena that remain unexplained. This paper reviews some of the recent findings on mitochondrial influences on the manner in which embryos develop and how their genotypes are inherited in mammals, with particular emphasis on the genetic bottleneck effect. Animal models carrying a mix of mtDNAs (heteroplasmic) have been produced by karyoplast and cytoplast transplantation to analyze the segregation patterns at different stages during embryogenesis, in fetuses and offspring. Comparisons performed between murine and bovine reveal interesting changes in segregation and replication of transplanted mtDNAs. We have recently obtained Bos indicus and Bos taurus fetuses and calves from embryos reconstructed using enucleated polymorphic oocytes of Bos taurus origin. These and other findings on mitochondrial biology will have important implications in determining the cytoplasmic genotype of clones and in the preservation of endangered breeds and species. (C) 1999 by Elsevier B.V.

Effects of feed restriction on yield, retail cuts and tissue composition of carcass of Saanen kids

Com o objetivo de estudar o efeito da restrição alimentar sobre as características de carcaça de caprinos leiteiros, realizou-se um experimento utilizando 27 cabritos machos Saanen distribuídos nos tratamentos 0 (alimentação à vontade), 30 ou 60% de restrição. Os animais apresentavam 5 kg de PV inicial e foram abatidos quando atingiram 20 kg de PV. Foram avaliados os rendimentos comercial e biológico, os cortes comerciais, a composição tecidual da perna, a área de olho-de-lombo (AOL) e a compacidade da carcaça. Utilizaram-se o delineamento inteiramente ao acaso e a análise de regressão em função da restrição alimentar. A restrição alimentar provocou redução do peso da carcaça e dos cortes comerciais, aumento da proporção do pescoço e diminuição da proporção do lombo. A proporção de ossos aumentou e a do tecido muscular e da gordura total diminuiu com o aumento da restrição. A proporção de gordura subcutânea diminuiu com o aumento da restrição alimentar. A área de olho-de-lombo (AOL) e a compacidade da carcaça foram afetadas pela restrição alimentar, mas ambas as medidas podem ser utilizadas para predizer a proporção de músculo da carcaça. O tratamento com 30% de restrição alimentar não prejudica a qualidade da carcaça de cabritos leiteiros.

Improving photoprotection of Beauveria bassiana conidia for biological control of the cactus pest Dactylopius opuntiae in the semiarid region northeast of Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

PHOSPHORUS EFFECTS IN THE NUTRITION and GROWTH of DEVELOPING MANGO PLANTS

The objective was to study the effects of phosphorous (P) fertilization on nutritional and developmental aspects of growing mango plants. The mango plants were evaluated by soil chemical analyses, leaf chemical analyses, biological examination of plant growth, and the starting point of fruit production. Having this in view, an experiment was set up on 2 January 2003, at Flora, a farm in Uberlandia, state of Minas Gerais, Brazil. The soil was a clayish Oxisol. The doses at planting were: D0 = zero, D1 = 40, D2 = 100, D3 = 200, and D4 = 300 g of P2O5 plant-1. These doses at the beginning of the second year were multiplied by 1.5 and at the beginning of the third year by 2.0 and applied to the plants. The fertilizer used in this experiment was triple superphosphate (44% of P2O5.). During August of 2004, 2005, and 2006, soil samples were taken at a depth of 20 cm in between the plant rows. Leaf samples were taken during August of 2004 and 2005 to determine macro- and micronutrient contents in the leaves. Plant stem diameter was measured during January of 2004 and 2005. Plant height and crown radius were measured during January of 2005 and fruit production in 2005 and 2006. Fertilizer applications increased the level of P in the soil but significantly influenced plant performance only after the second year. The effects of phosphorus on mango plants take place slowly leading to increments in plant stem diameter only at the third year. Fruit set was not influenced by phosphorous fertilization.

Ultrastructural analysis of the low level laser therapy effects on the lesioned anterior tibial muscle in the Gerbil

Low level laser therapy (LLLT) is known for its positive results but studies on the biological and biomodulator characteristics of the effects produced in the skeletal muscle are Still lacking. In this Study the effects of two laser dosages, 5 or 10 J/cm(2), on the lesioned tibial muscle were compared. Gerbils previously lesioned by 100 g load impact were divided into three groups: GI (n = 5) controls, lesion non-irradiated; GII (n = 5), lesion irradiated with 5 J/Cm(2) and GIII (n = 5), lesion irradiated with 10 J/cm(2), and treated for 7 consecutive days with a laser He-Ne (lambda = 633 rim). After intracardiac perfusion, the muscles were dissected and reduced to small fragments, post-fixed in 1% osmium tetroxide, dehydrated in increasing alcohol concentrations, treated with propylene oxide and embedded in Spurr resin at 60 degrees C. Ultrafine Cuts examined on a transmission electron microscope (Jeol 1010) revealed in the control GI group a large number of altered Muscle fibers with degenerating mitochondria, intercellular substance containing degenerating cell fragments and budding blood capillaries with Underdeveloped endothelial cells. However, groups GII and GIII showed muscle fibers with few altered myofibrils, regularly contoured mitochondria, ample intermembrane spaces and dilated mitochondrial crests. The clean intercellular Substance showed numerous collagen fibers and capillaries with multiple abluminal processes, intraluminal protrusions and several pinocytic vesicles in endothelial cells. it was concluded that laser dosages of 5 or 10 J/cm(2) delivered by laser He-Ne (lambda = 633 rim) during 7 consecutive days increase mitochondrial activity in muscular fibers, activate fibroblasts and macrophages and stimulate angiogenesis, thus suggesting effectivity of laser therapy tinder these experimental conditions. (C) 2009 Elsevier Ltd. All rights reserved.

In vivo assessment of DNA damage and protective effects of extracts from Miconia species using the comet assay and micronucleus test

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of isocoumarins isolated from Paepalanthus bromelioides on mitochondria: Uncoupling, and induction/inhibition of mitochondrial permeability transition

Isolated mitochondria may undergo uncoupling, and in presence of Ca2+ at different conditions, a mitochondrial permeability transition (MPT) linked to protein,thiol oxidation, and demonstrated by CsA-sensitive mitochondrial swelling; these processes may cause cell death either by necrosis or by apoptosis. Isocoumarins isolated from the Brazilian plant Paepalanthus bromelioides (Eriocaulaceae) paepalantine (9,10-dihydroxy-5,7-dimethoxy-1H-naptho(2,3c)pyran-1-one), 8,8'-paepalantine dimer, and vioxanthin were assayed at 1-50 mu M on isolated rat liver mitochondria, for respiration, MPT, protein thiol oxidation, and interaction with the mitochondrial membrane using 1,6-diphenyl-1,3,5-hexatriene (DPH). The isocoumarins did not significantly affect state 3 respiration of succinate-energized mitochondria; they did however, stimulate 4 respiration, indicating mitochondrial uncoupling. Induction of MPT and protein thiol oxidation were assessed in succinate-energized mitochondria exposed to 10 mu M Ca2+; inhibition of these processes was assessed in non-energized organelles in the presence of 300 mu M t-butyl hydroperoxide plus 500 mu M Ca2+. Only paepalantine was an effective MPT/protein thiol oxidation inducer, also releasing cytochrome c from mitochondria; the protein thiol oxidation, unlike mitochondrial swelling, was neither inhibited by CsA nor dependent on the presence of Ca2+. Vioxanthin was an effective inhibitor of MPT/protein thiol oxidation. All isocoumarins inserted deeply into the mitochondrial membrane, but only paepalantine dimer and vioxantin decreased the membrane's fluidity. A direct reaction with mitochondrial membrane protein thiols, involving an oxidation of these groups, is proposed to account for MPT induction by paepalantine, while a restriction of oxidation of these same thiol groups imposed by the decrease of membrane fluidity, is proposed to account for MPT inhibition by vioxanthin. (c) 2006 Published by Elsevier B.V..