A comparative study of protein and enzymatic activity in venoms of some common wasps (Hymenoptera: Vespidae) from São Paulo State

The Hymenoptera Aculeata venoms, with few exceptions, have been poorly studied and characterized. Nevertheless, they have raised increasing interest due to their medical importance, since accidents with these insects are fairly frequent in Brazil and may cause severe allergic reactions. The objectives of the present work were the quantitative characterization of the main allergenic enzymes present in the venom of the species Polybia paulista, Polybia ignobilis, Polistes simillimus, and Agelaia pallipes pallipes through biochemical assays for the determination of total protein content, as well as the level of the enzymatic activity of phospholipase, hyaluronidase, acid phosphatase and esterase. These results, in addition to providing biochemical knowledge about the venom of the species in question, also supply studies that allow phylogenetic inferences among them.

Enzymatic variability among venoms from different subspecies of Apis mellifera (Hymenoptera: Apidae)

The enzymatic variability was analyzed in venom extracts from bees reared in different colonies of the Africanized, A. m. ligustica and A. m. carnica subspecies. The implications of this variation focused on the biochemistry differentiation and immunogenicity of these venoms. The results showed the existence of a huge variability among the subspecies as well as among the colonies for three out of the six tested components - hyaluronidase, acid phosphatase and proteases - suggesting the utilization of these features as possible biochemical markers. Furthermore, although not statistically significant, it was found that the Africanized bee venom presented slightly higher levels of protein content and esterase activity, when compared to the other subspecies. If the esterase plays a role in the pain intensity caused by the sting, as suggested elsewhere, this might suggest a reason for a bigger algogenicity of this venom in relation to that of European bees. On the other hand, A. m. ligustica bees presented the highest levels of proteolytic and acid phosphatase activities, whose functions are not enlightened in Hymenoptera venoms. The A. m. carnica workers presented the highest hyaluronidase and the lowest acid phosphatase activity levels. The extremely variable results among colonies of the same subspecies and among subspecies, for the tested venom components, justify the absence of correlation between allergic reactions and tests with pooled venom.

Mensuração de hormônios andrógenos, estrógeno, fosfatase ácida prostática (PAP) e antígeno prostático específico (PSA) em cães adultos com próstata normal e hiperplásica

Prostatic diseases have been a common problem in middle age and older intact male dogs. Among these, benign prostatic hyperplasia (BHP) is the most frequent, age-related and hormonal-dependent condition of human and canine prostate. Blood samples were collected from 37 male intact dogs, tree years old dogs or more to determine androgens, estrogen, prostatic acid phosphatase (PAP) and prostatic specific antigen (PSA) according to histopathological aspects. Low levels of estrogen and high levels of prostatic specific antigen (PSA) were founded in dogs with BHP, respectively. Seric and urinary PAP levels were high in dogs with hyperplasia.

Localização histoquímica de algumas hidrolases no epidídimo do gerbilo

This study mainly showed that alkaline phosphatase expression had been present in the proximal regions of the epididymidis ductus of the gerbil which comprised the initial segment and proximal caput. The reactivities of acid phosphatase and ATPase were strong in the proximal and distal regions of the epididymidis ductus at the level of the apical cytoplasm and epithelium, except at the corpus level, a very thin isthmus located between the caput and cauda epididymidis, and as a general rule a low enzymatic reactive region of the epididymis of gerbil. SDH revealed also low activities in all the regions and regional structures of the duct, except into the luminal content formed by storaged spermatozoa, prior on the cauda level. The enzymes presented in the epididymis were correlated to some histophysiological roles such as the enzymatic mediation of endocytosis, secretion, absorption and active transport concerning to phosphatases and ATPase and a possible mitochondrial role of SDH could occur at the spermatozoa level in which the middle pieces were formed by a great amount of mitochondria.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5μm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Comparative morphology of the ileum of three species of Cephalotes (formicidae, myrmicinae)

The histology (H.E.), histochemistry (carbohydrates, proteins, collagen, and calcium), enzyme profile (ATPase and Acid phosphatase), and ultramorphology of the ileum of Cephalotes atratus, C. clypeatus and C. pusillus were studied in order to obtain some information about the similarities of these species. The relationship among these three species, as well as the results obtained for the wall, and the contents of this portion of the digestive tract were discussed. In C. clypeatus the ileum is somewhat smaller than that of C. atratus, despite the similarities in body size. The ATPase results showed 2 different regions: the anterior region responsible for absorbing material from the haemolymph and the posterior region for absorbtion from the ileum lumen. Other histochemical analyses failed to show any differences.

Efeito do alumínio no conteúdo de poliaminas livres e atividade da fosfatase ácida durante o crescimento de brotações de Eucalyptus grandis × E. urophylla cultivadas in vitro

This paper is aiming to evaluate the effects of different levels of the aluminum on the growth of the Eucalyptus grandis x E. urophylla shoots cultivated in vitro. Evaluations were carried out on pH and chemicals modifications of the culture medium by Geochem program, polyamines contents (putrescine, espermidine e espermine) and acid phosphatase ativity on the shoots. The trial had a totally randomized design with four treatments and four replicates. The treatments were: 0.0, 6.75,13.50 and 27 mg.L -1 of AlCl 3.6H 2. Evaluations were carried out on the 4th, 8th, 12th, 16th, 20th, 24th and 28th day of culture. The addition of the aluminium, in all concentrations, affected the culture medium ionic equilibrium, the morphology of the shoots, reduced the pH on the medium, induced an increase in polyamines content and higher acid phosphatase activity.

Programmed cell death in salivary glands of Drosophila arizonae and Drosophila mulleri

Programmed cell death (PCD) in insect metamorphosis assumes a great diversity of morphology and controlling processes that are still not well understood. With the objective of obtaining information about the PCD process, salivary glands of Drosophila arizonae and D. mulleri were studied during larval-pupal development. From the results, it can be concluded that the type of the PCD that occurs in these organs is morphologically typical of apoptosis (formation of apoptotic nuclei, followed by fragmentation into apoptotic bodies). Histolysis happens in both species, between 22 and 23 h after pupation. There were no significant differences between the species studied. Apoptosis does not occur simultaneously in all cells. Cytoplasmic acid phosphatase activity gradually increases during development, suggesting the existence of acid phosphatases that are only expressed during the apoptotic stage. Twenty hours after pupation, salivary glands already show biochemical alterations relative to nuclear permeability such as acidification, possibly due to the fusion of lysosomes with the nucleus a few hours before apoptosis. Autophagy seems to act together with apoptosis and has a secondary role in cell death. ©FUNPEC-RP.

Efeito de níveis de fósforo no crescimento inicial, biomassa e atividade in vivo da fosfatase ácida em Lychnophora ericoides Mart

The effect of phosphorus levels on seedling growth, dry and fresh matter production, and in vivo acid phosphatase activity was studied in Lychnophora ericoides cultivated during 130 days after transplanting in a greenhouse. Experimental design was completely randomized, with four P 2O 5 levels (0; 40; 80; 160 mg dm -3 as triple superphosphate) and five replicates of four plants each. The used soil was dystrophic acid dark red latosol. Lychnophora ericoides is responsive to phosphate fertilization with modification in daily relative growth rate, dry matter production and distribution among the plant parts. Acid phosphatase activity had significant and negative correlation with total, leaf and stem dry matter production, as well as with P levels accumulated in the leaf tissue. On the other hand, acid phosphatase activity had significant and positive correlation with root/shoot ratio.

Effects of treatment of the fat body trophocytes of Melipona quadrifasciata anthidioides nurse workers and virgin queens in culture by juvenile hormone III and ecdysterone (20-HE)

The fat body (FB) consists of two types of cells: throphocytes and oenocytes. Throphocytes are related to intermediary metabolism storing lipids, carbohydrates, and proteins while oenocytes play role in the lipids and lipoproteins production. The vitellogenin is the precursor of egg yolk (vitelline) and is synthesized on FB. The aim of this work was to analyze the effects of hormones acting in bee reproduction, as juvenile hormone (JH) and ecdisteroids (20 HE) on FB cells, where vitellogenin is synthesized. For the study were chose nurse workers that in Melipona quadrifasciata anthidioides present activated ovaries and produce eggs, and virgin queens whose ovaries are not yet activated, presenting only previtellogenic follicles. FB trophocytes from these classes of bees were cultivated in media containing different amounts of JH and 20-HE. The effects on trophocytes cytoplasm reserves of lipids, proteins, and activity of acid phosphatase were compared by observing preparations from cultured FB, treated and control, by transmission electron microscopy (TEM). The results showed that the hormones effects are related to the bee's caste and functional ovary stage. The role of acid phosphatase on mobilization of the trophocyte reserves was also determined. © 2012 Wiley Periodicals, Inc.

Neurotoxic action of permethrin in Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) female ticks. Morphological and cytochemical evaluation of the central nervous system

Studies on the molecular bases of the neurotoxic action of acaricides are found in the literature; but there are no studies of this action on the nervous system of ticks at the cellular level. The present study describes the morphological and cytochemical changes in the synganglion of Rhipicephalus sanguineus semi-engorged females exposed to different concentrations of permethrin, a pyrethroid with recognized neurotoxic action. Permethrin induced the degeneration of the synganglion through a process of apoptosis involving autophagy, characterized by the condensation and margination of the chromatin, formation of blebs in the nuclear envelope and fragmentation of the nucleus, loss of shape of neural cells and integrity of cellular membrane, cytoplasmic shrinkage, and lower levels of acid phosphatase in the nervous tissue as the concentration of permethrin increased. This study provided further evidence of the neurotoxic action of permethrin, which impairs the metabolism of R. sanguineus nervous systems, and consequently the physiology of other systems, dependent on the neural control. These results provide cytochemical and histological confirmation of the neurotoxic action of permethrin, previously inferred from molecular and tick behavioral evidence. © 2013 Elsevier B.V.

Fish exposure to nano-TiO2 under different experimental conditions: Methodological aspects for nanoecotoxicology investigations

The ecotoxicology of nano-TiO2 has been extensively studied in recent years; however, few toxicological investigations have considered the photocatalytic properties of the substance, which can increase its toxicity to aquatic biota. The aim of this work was to evaluate the effects on fish exposed to different nano-TiO2 concentrations and illumination conditions. The interaction of these variables was investigated by observing the survival of the organisms, together with biomarkers of biochemical and genetic alterations. Fish (Piaractus mesopotamicus) were exposed for 96h to 0, 1, 10, and 100mg/L of nano-TiO2, under visible light, and visible light with ultraviolet (UV) light (22.47J/cm2/h). The following biomarkers of oxidative stress were monitored in the liver: concentrations of lipid hydroperoxide and carbonylated protein, and specific activities of superoxide dismutase, catalase, and glutathione S-transferase. Other biomarkers of physiological function were also studied: the specific activities of acid phosphatase and Na,K-ATPase were analyzed in the liver and brain, respectively, and the concentration of metallothionein was measured in the gills. In addition, micronucleus and comet assays were performed with blood as genotoxic biomarkers. Nano-TiO2 caused no mortality under any of the conditions tested, but induced sublethal effects that were influenced by illumination condition. Under both illumination conditions tested, exposure to 100mg/L showed an inhibition of acid phosphatase activity. Under visible light, there was an increase in metallothionein level in fish exposed to 1mg/L of nano-TiO2. Under UV light, protein carbonylation was reduced in groups exposed to 1 and 10mg/L, while nucleus alterations in erythrocytes were higher in fish exposed to 10mg/L. As well as improving the understanding of nano-TiO2 toxicity, the findings demonstrated the importance of considering the experimental conditions in nanoecotoxicological tests. This work provides information for the development of protocols to study substances whose toxicity is affected by illumination conditions. © 2013 Elsevier B.V..

Alterações morfofisiológicas do tegumento de coelhos infestados por carrapatos Rhipicephalus sanguineus (Acari: Ixodidae) e expostos à selamectina (princípio ativo do acaricida Revolution®, Pfizer)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo morfológico comparativo das glândulas salivares de fêmeas de carrapatos Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae) em dois estágios de alimentação

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Atividade de fosfomonohidrolases envolvidas com o crescimento e absorção da cauda de girinos de rã-touro (Lithobates catesbeianus)

Pós-graduação em Biotecnologia - IQ