Pollen analysis of honey and beebread derived from Brazilian mangroves

Pollen analyses were performed on honey and beebread from hives in apiaries located in two distinct mangrove areas dominated by Laguncularia racemosa (L.) C.F. Gaernt. One apiary was located at the edge of Guanabara Bay, Rio de Janeiro State, and the other near Maranguá Bay, Bahia State, Brazil. We investigated the contribution of nectar and pollen from mangrove vegetation to Apis mellifera L. honey and beebread stocks. Intensive visitation to this plant species by honeybees and the presence of its pollen grains in honey and beebread confirmed the importance of Laguncularia racemosa as a polliniferous and nectariferous species.

Single-step purification of crotapotin and crotactine from Crotalus durissus terrificus venom using preparative isoelectric focusing

We describe the isolation of crotoxin, a presynaptic B-neurotoxin, as well as its subunits B (crotactine) and A (crotapotin) from lyophilized Crotalus durissus terrificus venom by a single-step preparative isoelectric focusing procedure. From 98 mg of dried venom protein 20.1 mg of crotactine and 13.1 mg of crotapotin were recovered in the first step of focalization and 4.2 mg in a second run. These values correspond to 35.7% of the total venom protein applied. Crotactine separated in the 9.3-7.0 pH range (tubes 1-6) and crotapotin in the 1.8-2.8 pH range (tubes 15-19) and both were homogeneous by SDS-PAGE and N-terminal amino acid analysis. Crotactine, a 12-kDa protein, presented hemolytic and phospholipase A2 activity. Thus, using isoelectric focusing we simultaneously purified both toxins in high yields. This method can be used as an alternative for the purification and characterization of proteins from other snake venoms under conditions in which biological activity is retained

Ethological analysis of mother-pup interactions and other behavioral reactions in rats: effects of malnutrition and tactile stimulation of the pups

Mother-pup interaction, as well as other behavioral reactions were studied during the lactation period in 24 litters of Wistar rats and their dams fed either a 16% (control - C; 12 litters) or a 6% (malnourished - M; 12 litters) protein diet. The diets were isocaloric. Throughout lactation there was a 36.4% weight loss of M dams and a 63% body weight deficit in the M pups when compared to control pups. During this period, half of the litters were exposed daily to additional tactile stimulation (CS or MS), while the other half were submitted to normal rearing conditions (CN or MN). The tactile stimulation of pups (handling) consisted of holding the animal in one hand and gently touching the dorsal part of the animal's body with the fingers for 3 min. A special camera and a time-lapse video were used to record litter behavior in their home cages. Starting at 6 p.m. and ending at 6 a.m., on days 3, 6, 12, 15, 18 and 21 of lactation, photos were taken at 4-s intervals. An increase in the frequency (154.88 ± 16.19) and duration (455.86 ± 18.05 min) of suckling was observed throughout the lactation period in all groups compared to birth day (frequency 24.88 ± 2.37 and duration 376.76 ± 21.01 min), but the frequency was higher in the C (84.96 ± 8.52) than in the M group (43.13 ± 4.37); however, the M group (470.2 ± 11.87 min) spent more time suckling as compared with the C group (393.67 ± 13.09 min). The M dams showed a decreased frequency of resting position throughout the lactation period (6.5 ± 2.48) compared to birth day (25.42 ± 7.74). Pups from the C group were more frequently observed separated (73.02 ± 4.38) and interacting (258.99 ± 20.61) more with their mothers than the M pups (separated 66.94 ± 5.5 and interacting 165.72 ± 12.05). Tactile stimulation did not interact with diet condition, showing that the kind of stimulation used in the present study did not lead to recovery from the changes induced by protein malnutrition. The changes in mother-pup interaction produced by protein malnutrition of both may represent retardation in neuromotor development and a higher dependence of the pups on their mothers. These changes may represent an important means of energy saving and heat maintenance in malnourished pups.

Comparison of the specificity of PCR and the histopathological detection of leishmania for the diagnosis of American cutaneous leishmaniasis

More precise and rapid diagnostic methods for American cutaneous leishmaniasis (ACL) are necessary because of the growing number of cases observed in Brazil, including the northeastern region of the State of São Paulo. We applied PCR to 54 skin or mucosal biopsies from patients with a clinical and/or laboratory diagnosis of ACL using primers 13A and 13B, with positive results being obtained for 82% of the samples. When the PCR results were compared to those of histopathological leishmania detection, PCR showed superior results with 81.5% sensitivity and 95% CI of 68.0-95.1%. The Montenegro skin test (MST) was positive in 88.7% of patients. Since MST cannot be used as a diagnostic tool in endemic areas, the present results strongly suggest the use of PCR for the etiological confirmation of ACL, with emphasis on the mucosal form.

Anti-inflammatory and antispasmodic activity of Ipomoea imperati (Vahl) Griseb (Convolvulaceae)

Ipomoea imperati (Convolvulaceae) lives on the sandy shores of the Brazilian coast and in other areas of the world. The anti-inflammatory activity of a methanol-water extract of the leaves of I. imperati was investigated in experimental models of acute and subchronic inflammation. Topical application of the extract (10 mg/ear) inhibited mouse ear edema induced by croton oil (89.0 ± 1.3% by the lipid fraction with an IC50 of 3.97 mg/ear and 57.0 ± 1.3% by the aqueous fraction with an IC50 of 3.5 mg/ear) and arachidonic acid (42.0 ± 2.0% with an IC50 of 4.98 mg/ear and 31.0 ± 2.0% with an IC50 of 4.72 mg/ear). Phospholipase A2, purified from Apis mellifera bee venom, was also inhibited by the extract (5.0 mg/ml lipid and aqueous fraction) in vitro in a dose-dependent manner (85% by the lipid fraction with an IC50 of 3.22 mg/ml and 25% by the aqueous fraction with an IC50 of 3.43 mg/ml). The methanol-water extract of I. imperati (1000 mg/kg) administered by the oral route also inhibited the formation of cotton pellet-induced granulomas (73.2 ± 1.2% by the lipid fraction and 56.14 ± 2.7% by the aqueous fraction) and did not cause gastric mucosal lesions. I. imperati extracts (10 mg/ml) also inhibited in a dose-dependent manner the muscle contractions of guinea pig ileum induced by acetylcholine and histamine (IC50 of 1.60 mg/ml for the lipid fraction and 4.12 mg/ml for the aqueous fraction). These results suggest the use of I. imperati as an anti-inflammatory and antispasmodic agent in traditional medicine.

Evaluation of stunned and infarcted canine myocardium by real time myocardial contrast echocardiography

Differentiation between stunned and infarcted myocardium in the setting of acute ischemia is challenging. Real time myocardial contrast echocardiography allows the simultaneous assessment of myocardial perfusion and function. In the present study we evaluated infarcted and stunned myocardium in an experimental model using real time myocardial contrast echocardiography. Sixteen dogs underwent 180 min of coronary occlusion followed by reperfusion (infarct model) and seven other dogs were submitted to 20 min of coronary occlusion followed by reperfusion (stunned model). Wall motion abnormality and perfusional myocardial defect areas were measured by planimetry. Risk and infarct areas were determined by tissue staining. In the infarct model, the wall motion abnormality area during coronary occlusion (5.52 ± 1.14 cm²) was larger than the perfusional myocardial defect area (3.71 ± 1.45 cm²; P < 0.001). Reperfusion resulted in maintenance of wall motion abnormality (5.45 ± 1.41 cm²; P = 0.43 versus occlusion) and reduction of perfusional myocardial defect (1.51 ± 1.29 cm²; P = 0.004 versus occlusion). Infarct size determined by contrast echocardiography correlated with tissue staining (r = 0.71; P = 0.002). In the stunned model, the wall motion abnormality area was 5.49 ± 0.68 cm² during occlusion and remained 5.1 ± 0.63 cm² after reperfusion (P = 0.07). Perfusional defect area was 2.43 ± 0.79 cm² during occlusion and was reduced to 0.2 ± 0.53 cm² after reperfusion (P = 0.04). 2,3,5-Triphenyl tetrazolium chloride staining confirmed the absence of necrotic myocardium in all dogs in the stunned model. Real time myocardial contrast echocardiography is a noninvasive technique capable of distinguishing between stunned and infarcted myocardium after acute ischemia.

Comparison of scientists of the Brazilian Academy of Sciences and of the National Academy of Sciences of the USA on the basis of the h-index

A new scientometric indicator, the h-index, has been recently proposed (Hirsch JE. Proc Natl Acad Sci 2005; 102: 16569-16572). The index avoids some shortcomings of the calculation of the total number of citations as a parameter to evaluate scientific performance. Although it has become known only recently, it has had widespread acceptance. A comparison of the average h-index of members of the Brazilian Academy of Sciences (BAS) and of the National Academy of Sciences of the USA (NAS-USA) was carried out for 10 different areas of science. Although, as expected, the comparison was unfavorable to the members of the BAS, the imbalance was distinct in different areas. Since these two academies represent, to a significant extent, the science of top quality produced in each country, the comparison allows the identification of the areas in Brazil that are closer to the international stakeholders of scientific excellence. The areas of Physics and Mathematics stand out in this context. The heterogeneity of the h-index in the different areas, estimated by the median dispersion of the index, is significantly higher in the BAS than in the NAS-USA. No elements have been collected in the present study to provide an explanation for this fact.

Increased expression of keratinase and other peptidases by Candida parapsilosis mutants

Keratinases are enzymes of great importance involved in pathogenic processes of some fungi. They also have a widespread ecological role since they are responsible for the degradation and recycling of keratin. On the one hand, studying them furthers our knowledge of pathogenicity mechanisms, which has important implications for human health, and on the other hand, understanding their ecological role in keratin recycling has biotechnological potential. Here, a wild-type keratinolytic Candida parapsilosis strain isolated from a poultry farm was treated with ethyl methanesulfonate in order to generate mutants with increased keratinase activity. Mutants were then cultured on media with keratin extracted from chicken feathers as the sole source of nitrogen and carbon. Approximately 500 mutants were screened and compared with the described keratinolytic wild type. Three strains, H36, I7 and J5, showed enhanced keratinase activity. The wild-type strain produced 80 U/mL of keratinolytic activity, strain H36 produced 110 U/mL, strain I7, 130 U/mL, and strain J5, 140 U/mL. A 70% increase in enzyme activity was recorded for strain J5. Enzymatic activity was evaluated by zymograms with proteic substrates. A peptidase migrating at 100 kDa was detected with keratin, bovine serum albumin and casein. In addition, a peptidase with a molecular mass of 50 kDa was observed with casein in the wild-type strain and in mutants H36 and J5. Gelatinase activity was detected at 60 kDa. A single band of 35 kDa was found in wild-type C. parapsilosis and in mutants with hemoglobin substrate.

HYBRID ARTIFICIAL NEURAL NETWORK APPLIEDTO MODELING SCFE OF BASIL AND ROSEMARY OILS

This work presents the results of a Hybrid Neural Network (HNN) technique as applied to modeling SCFE curves obtained from two Brazilian vegetable matrices. A series Hybrid Neural Network was employed to estimate the parameters of the phenomenological model. A small set of SCFE data of each vegetable was used to generate an extended data set, sufficient to train the network. Afterwards, other sets of experimental data, not used in the network training, were used to validate the present approach. The series HNN correlates well the experimental data and it is shown that the predictions accomplished with this technique may be promising for SCFE purposes.

Effect of pre and post-slaughter processes on meat characteristics of Santa Ines ewes discarded due to age

The effects of body condition recovery (BC), carcass electrical stimulation (ES), aging time (AT 7 - 14 days), and calcium chloride injection on the meat characteristics of Santa Inês ewes (±5 years old) slaughtered immediately after weaning or after the body condition recovery period were studied. The carcass temperature, pH, shear force (SF), cooking loss (CL), meat color (L*, a*, b*), and meat tenderness were evaluated. A completely randomized design in a 2 × 2 × 2 × 3 (BC × ES × CaCl2 × AT) factorial arrangement was used, and the sensory tenderness data were analyzed using the table of Minimum Number of Correct Answers for the Duo-Trio test. The body condition recovery reduces the shear force in 8%, increasing their tenderness. Electrical stimulation reduced the shear force (24%) and did not change the other parameters. The aging time (7 or 14 days) decreased the shear force (18-26%), effect that was enhanced by electrical stimulation, and it darkened the meat reducing lightness (L*) and increasing yellowness (b*). The treatment with CaCl2 was the most effective in tenderizing meat by reducing the shear force ( 35%); increasing the cooking loss (4.5%); and increasing L* and b* lightening the meat. The sensory evaluation of tenderness corroborates the findings of the experimental evaluation regarding the effect of the treatment with CaCl2 on the meat quality improvement. It was concluded that the treatments improve meat characteristics achieving better results when applied together.

Phylogenetic Analysis of Avidins and Expression of Novel Avidins from Lactrodectus hesperus and Hoeflea phototrophica

Avidins (Avds) are homotetrameric or homodimeric glycoproteins with typically less than 130 amino acid residues per monomer. They form a highly stable, non-covalent complex with biotin (vitamin H) with Kd = 10-15 M (for chicken Avd). The best-studied Avds are the chicken Avd from Gallus gallus and streptavidin from Streptomyces avidinii, although other Avd studies have also included Avds from various origins, e.g., from frogs, fishes, mushrooms and from many different bacteria. Several engineered Avds have been reported as well, e.g., dual-chain Avds (dcAvds) and single-chain Avds (scAvds), circular permutants with up to four simultaneously modifiable ligand-binding sites. These engineered Avds along with the many native Avds have potential to be used in various nanobiotechnological applications. In this study, we made a structure-based alignment representing all currently available sequences of Avds and studied the evolutionary relationship of Avds using phylogenetic analysis. First, we created an initial multiple sequence alignment of Avds using 42 closely related sequences, guided by the known Avd crystal structures. Next, we searched for non-redundant Avd sequences from various online databases, including National Centre for Biotechnology Information and the Universal Protein Resource; the identified sequences were added to the initial alignment to expand it to a final alignment of 242 Avd sequences. The MEGA software package was used to create distance matrices and a phylogenetic tree. Bootstrap reproducibility of the tree was poor at multiple nodes and may reflect on several possible issues with the data: the sequence length compared is relatively short and, whereas some positions are highly conserved and functional, others can vary without impinging on the structure or the function, so there are few informative sites; it may be that periods of rapid duplication have led to paralogs and that the differences among them are within the error limit of the data; and there may be other yet unknown reasons. Principle component analysis applied to alternative distance data did segregate the major groups, and success is likely due to the multivariate consideration of all the information. Furthermore, based on our extensive alignment and phylogenetic analysis, we expressed two novel Avds, lacavidin from Lactrodectus Hesperus, a western black widow spider, and hoefavidin from Hoeflea phototrophica, an aerobic marine bacterium, the ultimate aim being to determine their X-ray structures. These Avds were selected because of their unique sequences: lacavidin has an N-terminal Avd-like domain but a long C-terminal overhang, whereas hoefavidin was thought to be a dimeric Avd. Both these Avds could be used as novel scaffolds in biotechnological applications.

The role of nuclear and other conventional power plants in the flexible energy system

This thesis reviews the role of nuclear and conventional power plants in the future energy system. The review is done by utilizing freely accesible publications in addition to generating load duration and ramping curves for Nordic energy system. As the aim of the future energy system is to reduce GHG-emissions and avoid further global warming, the need for flexible power generation increases with the increased share of intermittent renewables. The goal of this thesis is to offer extensive understanding of possibilities and restrictions that nuclear power and conventional power plants have regarding flexible and sustainable generation. As a conclusion, nuclear power is the only technology that is able to provide large scale GHG-free power output variations with good ramping values. Most of the currently operating plants are able to take part in load following as the requirement to do so is already required to be included in the plant design. Load duration and ramping curves produced prove that nuclear power is able to cover most of the annual generation variation and ramping needs in the Nordic energy system. From the conventional power generation methods, only biomass combustion can be considered GHG-free because biomass is considered carbon neutral. CFB combusted biomass has good load follow capabilities in good ramping and turndown ratios. All the other conventional power generation technologies generate GHG-emissions and therefore the use of these technologies should be reduced.

Influence of disinfestation and osmotic conditioning on the germinating behavior of australian royal palm (Archontophoenix alexandrae)seeds

The technique of Osmotic Conditioning, which consists of partial and controlled hydration of the seeds, has obtained success with various species of seeds, increasing the germinating span and tolerance to the adverse conditions of the environment, and has also reduced the time elapsed between sowing and the emergence of the plants. Associated to ideal storage conditions, the treatment has increased the performance of the seeds of tropical wood species. Aiming at studying the germinating environment and the effect of osmotic conditioning on the germination of seeds of the Australian Royal Palm tree, two experiments were performed. The first one evaluated the effect of disinfestation of the seeds of the Australian Royal Palm tree with NaClO. The treatments applied were: 0.5% sodium hypochlorite, exposure periods of 5, 15, 30, 45, 60, 90, 120 and 240 minutes, and the fungicide Captan, as control. The treatments with NaClO did not differ in relation to the final percentage of germination and to the germination speed index, and did not differ from the treatment control. The second test evaluated solutions with the following osmotic potentials: 0.0MPa (pure water), -0.4MPa, -0.6MPa and -0.8MPa, exposed for the periods of 10 and 20 days. The final percentage of germination did not differ among the treatments. The seeds hydrated in pure water for a period of 20 days showed a germination speed index significantly superior to the other treatments, and they did not show significant differences among themselves.

Marks of Fire, Value and Faith. Swords with Ferrous Inlays in Finland during the Late Iron Age (ca. 700–1200 AD)

This study explores swords with ferrous inlays found in Finland and dating from the late Iron Age, ca. 700–1200 AD. These swords reflect profound changes not only in styles and fashion but also in the technology of hilts and blades. This study explores how many of these kinds of swords are known from Finland, how they were made and where, what their status was in Late Iron Age Finland, and where the Finnish finds stand in accordance with other areas of Europe. The various methods included measuring of the finds and statistics. The main method of revealing the inlaid marks was radiography due to its non-destructive nature. In cases where inlays were visible without radiography, their details were inspected via microscopy. To study the materials and manufacture of inlaid swords, a sample of them was metallographically analysed to determine the forging technologies and nature of used materials. Furthermore, the manufacture was also studied with experimental approaches. As a result, a catalogue of 151 swords with ferrous inlays was created. This number is relatively high compared with other European countries, although systematic studies have been conducted in only some countries. The inlaid motifs were classified into five distinct categories to help the classification. To summarize, almost every documented inlaid sword was unique in some respect including measurements, inlaid motifs and materials of blades and inlays. Technological variation was also present, some blades being poorer and some of higher quality in spite of the inlaid motifs. Misspelt inscriptions as well as letter-like marks were common in Finland and also in Scandinavia. Furthermore, the provenance of iron and steel used in some blades hints at Scandinavian ores. The above observations, along with the experimental results indicating the existence of multiple alternative techniques of inlaying, suggest that these swords were manufactured locally in Scandinavia, most likely in imitation of Continental European models. Inlaid swords were valued partly for their assumed functionality in combat, as evidenced by damage on some examined blades, or they were valued for their inlays, which could have had fashionable or symbolical meanings bound to local beliefs.

A critical examination of the involvement of Canadian high-performance athletes in the development of anti-doping policy

The use of certain perfonnance enhancing substances and methods has been defined as a major ethical breach by parties involved in the governance of highperfonnance sport. As a result, elite athletes worldwide are subject to rules and regulations set out in international and national anti-doping policies. Existing literature on the development of policies such as the World Anti-Doping Code and The Canadian antiDoping Program suggests a sport system in which athletes are rarely meaningfully involved in policy development (Houlihan, 2004a). Additionally, it is suggested that this lack of involvement is reflective of a similar lack of involvement in other areas of governance concerning athletes' lives. The purpose ofthis thesis is to examine the history and current state of athletes' involvement in the anti-doping policy process in Canada's high-perfonnance sport system. It includes discussion and analysis of recently conducted interviews with those involved in the policy process as well as an analysis of relevant documents, including anti-doping policies. The findings demonstrate that Canadian athletes have not been significantly involved in the creation of recently developed antidoping policies and that a re-evaluation of current policies is necessary to more fully recognize the reality of athletes' lives in Canada's high-perfonnance sport system and their rights within that system.