Antitumor effects in vitro and in vivo and mechanisms of protection against melanoma B16F10-Nex2 cells by fastuosain, a cysteine proteinase from Bromelia fastuosa

In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment ( mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein - chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies.

Triiodothyronine mimics the effects of estrogen in breast cancer cell lines

MCF-7 (estrogen receptor positive - ER(+)) and MDA-MB-231 (estrogen receptor negative - ER(-)) are human breast cancer cell lines which express functional thyroid hormone receptors (c-erb A alpha 1 and c-erb beta 1) as indicated by stimulation of mitochondrial alpha-glycerophosphate dehydrogenase. In MCF-7, mimicking E(2), T-3 stimulated growth in a dose-dependent (10(10) M-10(-8) M) manner, induced the expression of progesterone receptor and growth factor TGF alpha mRNAs and inhibited that of TGF beta mRNA; T-3 also increased progesterone binding and LDH5 isozyme activities. None of these effects were observed in (ER(-)) MDA-MB-231 cells. 10(-6) M tamoxifen (TAM) reverted growth stimulation, suppressed progesterone receptor and TGF alpha mRNA induction and restored TGF beta mRNA to control levels in T-3-treated MCF-7 cells. That T-3 is acting in MCF-7 cells via its binding to ER is suggested by the immunoprecipitation of pre-bound I-125-T-3 from MCF-7 nuclear extracts by an ER-specific monoclonal antibody and by the displacement of H-3-estradiol binding to ER by radioinert T-3. Copyright (C) 1996 Elsevier B.V. Ltd.

Effects of recovery type after a judo combat on blood lactate removal and on performance in an intermittent anaerobic task

Aim. The objective of this study was to verify the effects of active (AR) and passive recovery (PR) after a judo match on blood lactate removal and on performance in an anaerobic intermittent task (4 bouts of upper body Wingate tests with 3-min interval between bouts; 4WT).Methods. The sample was constituted by 17 male judo players of different competitive levels: A) National (Brazil) and International medallists (n. 5). B) State (São Paulo) medallists (n. 7). Q City (São Paulo) medallists (n. 5). The subjects were submitted to: 1) a treadmill test for determination of VO2peak and velocity at anaerobic threshold (VAT); 2) body composition; 3) a 5-min judo combat, 15-min of AR or PR followed by 4WT.Results. The groups did not differ with respect to: body weight, VO2peak, VAT, body fat percentage, blood lactate after combats. No difference was observed in performance between AR and PR, despite a lower blood lactate after combat (10 and 15 min) during AR compared to PR. Groups A and B performed better in the high-intensity intermittent exercise compared to athletes with lower competitive level (C).Conclusion. The ability to maintain power output during intermittent anaerobic exercises can discriminate properly judo players of different levels. Lactate removal was improved with AR when compared to PR but AR did not improve performance in a subsequent intermittent anaerobic exercise.

Novel allosteric conformation of human HB revealed by the hydration and anion effects on O-2 binding

The effect of anions on the stability of different functional conformations of Hb is examined through the determination of the dependence of O-2 affinity on water activity (a(w)). The control of a(w) is effected by varying the sucrose osmolal concentration in the bathing solution according to the osmotic stress method. Thus, the hydration change following Hb oxygenation is determined as a function of Cl- and of DPG concentration. We find that only similar to 25 additional water molecules bind to human Hb during the deoxy-to-oxy conformation transition in the absence of anions, in contrast with similar to 72 that bind in the presence of more than 50 mM Cl- or more than 15 mu M DPG. We demonstrate that the increase in the hydration change linked with oxygenation is coupled with anion binding to the deoxy-Hb. Hence, we propose that the deoxy-Hb coexists in two allosteric conformations which depend on whether anion is bound or not: the tense T-state, with low oxygen affinity and anion bound, or a new allosteric P-state, with intermediate oxygen affinity and free of bound anions. The intrinsic oxygen affinity of this unforeseen P-state and the differential binding of Cl-, DPG, and H2O between states P and T and P and R are characteristics which are consistent with those expected for a putative intermediate allosteric state of Hb. These findings represent a new opportunity to explore the structure-function relationships of hemoglobin regulation.

Well productivity controlling factors in crystalline terrains of southeastern Brazil

Over the last decades, increasing water demands have fostered research to obtain high well yields in crystalline terrains where, besides the intrinsic properties of rocks, the groundwater flow depends on several factors. The depth of the wells, the lithotypes, the presence and thickness of sedimentary coverings and weathered layers, the landforms, the geological structures, and the effects of tectonic stresses are among the most investigated factors considered as determinant of well productivity. The influence of these factors on productivity of wells that exploit the Crystalline Aquifer System in the Jundiai-River Catchment, southeastern Brazil, is investigated in this work. The largest region of the studied area is located on the Precambrian Basement, partially covered by sedimentary deposits. The results show that the sedimentary deposits and the weathered layer are important for high well yield, but it also depends on the existence of a net of open fractures, in order to maintain high productivity. The sites that have more possibility of occurrence of such structures are the regional shear and fault zones and other minor structures with NW-SE and E-W directions, which characterize areas subjected to transtensional stress related to the neotectonics.

Antinociceptive effects of epidural buprenorphine or medetomidine, or the combination, in conscious cats

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

Effects of a temperature dependent viscosity in surface nonlinear waves propagating in a shallow fluid heated from below

The effects of a temperature dependent viscosity in surface nonlinear waves propagating in a shallow fluid heated from below are investigated. It is shown that the (2+1)-dimensional Burgers equation may appear as the equation governing the upper free surface perturbations of a Bénard system, even when the viscosity is assumed to depend on temperature. The critical Rayleigh number for the appearance of waves governed by the Kadomtsev-Petviashvili equation, however, will be smaller than R=30, which is the critical number obtained for a constant viscosity. © 1992.

Effects of tunicamycin on glycoprotein antigens of Aspergillus fumigatus

Tunicamycin, which inhibits N-glycosylation of proteins, was used as a tool to determine the type of linkage which occurs in glycoprotein antigens of Aspergillus fumigatus. When A. fumigatus extracts were electrophoretically separated and blotted then probed with anti-Aspergillus patients' sera, differences in antigenic profiles were noted when tunicamycin-treated samples were compared with controls. Tunicamycin had no detectable effect on the cellular proteinases of A. fumigatus, most of which are glycosylated. Some enzymatic components were lacking when extracellular proteinases were compared with those of control samples. The major catalase component of A. fumigatus is a concanavalin A (Con A)-binding glycoprotein. In cultures grown in the presence of tunicamycin, partiallydeglycosylated catalase components were obtained which could be distinguished from the native catalase by their altered mobilities in polyacrylamide gels. The effect of deglycosylation on catalase antigens was monitored using an antiserum raised to a ConA-binding fraction of A fumigatus mycelium. These antibodies bound both to the native glycoprotein and the partially deglycosylated material. These latter two were largely unaffected when incubated with an antiserum raised to a non-ConA-binding fraction of A. fumigatus which is essentially carbohydrate free. The ability to produce partially-glycosylated antigens of A. fumigatus offers a model to study the effect of basic structural modifications on both the enzymatic and antigenic activities of these molecules.

Effects of fibrin adhesive material (Tissucol) on alveolar healing in rats under stress.

The effects of Tissucol on alveolar healing following stress were evaluated histologically, comparing three groups of 28 male albino rats each. Stress was applied and their right upper incisors were extracted. Group A served as an empty control site. In Group B, Tissucol was applied into the alveolar cavity. Group C received local antifibrinolytic treatment (alveolar irrigation with epsilon-aminocaproic acid solution) before implant of Tissucol into the tooth socket. Four animals in each group were killed at 1, 3, 6, 9, 15, 21 and 24 days after surgery. Results showed that: 1) Tissucol did not interfere with connective and osseous tissue formation; 2) Tissucol allowed new bone formation; 3) Tissucol residues in Group B in sections of 24-day specimens did not impair healing; 4) Tissucol was usually completely resorbed and healing was complete 24 days after surgery in Group C.

Effects of Chronic Physical Activity and of Ultrasound Treatment on Bone Consolidation

The objective of this study was to investigate the effects of ultrasound treatment and physical exercise on the velocity of bone consolidation and resistance to deformation. We performed osteotomy in the upper third of the right tibia of rats. Physical training consisted of swimming 1 h per day with a load of 5% b.w. for 30 days. Therapy with medium-intensity ultrasound was applied daily on the damaged area. Wistar rats were divided into the following groups: osteotomized sedentary animals with no ultrasound treatment (1.OSnUS), osteotomized trained animals with no ultrasound treatment (2.OTnUS), osteotomized sedentary animals with ultrasound treatment (3.OSwUS). and osteotomized trained animals with ultrasound treatment (4.OTwUS). The animals were sacrificed for the following analyses: muscle glycogen, serum alkaline phosphatase at the 5th, 10th, 20th, and 30th days, test of maximum resistance to flexion, rupture flexion and mean tibial rigidity at the 30th day. Muscle glycogen was increased at the 20th day: alkaline phosphatase was elevated at the 5th and 20th days in groups 3.OSwUS and 4.OTwUS. and decreased at the 10th day. Groups1.OSnUS and 2.OTnUS did not show significant variations. In the mechanical resistance tests, we noted that ultrasound therapy and the association of physical activity used in the present study showed significant differences in bone resistance and bone rigidity after 30 days of treatment. These facts suggest that ultrasound or physical activity, or their combination may accelerate the process of bone tissue repair.

Effect of hydrated sodium calcium aluminosilicate on the prevention of the toxic effects of ochratoxin

We studied the efficacy of hydrated sodium calcium aluminosilicate (HSCAS) as an inhibitor of the toxic effects of ochratoxin in broilers from 1 to 42 d of age. A total of 288 broilers was distributed into four treatments with four replicates of 18 birds each: T1, control; T2, 0.25% HSCAS; T3, 2 ppm of ochratoxin; T4, 0.25% HSCAS + 2 ppm ochratoxin. The parameters evaluated were feed intake; weight gain; feed conversion; relative weights of the liver, kidneys, and bursa; and serum levels of Ca, P, total protein (TP), aspartate aminotransferase (AST) and γ-glutamiltransferase (GGT). Ochratoxin in the diet negatively affected (P < 0.05) all performance parameters evaluated when the birds were 21 and 42 d of age. However, HSCAS did not affect performance, and there was no interaction between HSCAS and dietary ochratoxin. The liver and the kidneys of birds fed ochratoxin with or without HSCAS were relatively heavier (P < 0.05) than those of the control birds, demonstrating the influence of ochratoxin, but not of HSCAS, on the relative weight of these organs. Although the bursa of birds exposed to ochratoxin with or without HSCAS had a lower relative weight as compared to control birds, the difference was not significant. Ca, P, and TP serum levels were lower (P < 0.05) in birds exposed to ochratoxin, whereas AST and GGT levels were higher (P < 0.05) in these birds. These results reflect that ochratoxin in the diet impaired the productivity indexes and that HSCAS did not improve these parameters.

Treatment Effects Produced by Fränkel Appliance in Patients with Class II, Division 1 Malocclusion

The purpose of this investigation was to evaluate the dentoalveolar and skeletal cephalometric changes produced by the Fränkel appliance in individuals with a Class II, division 1 malocclusion. Lateral cephalograms of 44 patients of both sexes were divided in two groups of 22 each. The control group was comprised of untreated Class II children with an initial mean age of eight years and seven months who were followed without treatment for a period of 13 months. The Fränkel group had an initial mean age of nine years and was treated for a mean period of 17 months. Lateral cephalometric headfilms of each patient were obtained at the beginning and end of treatment. The Fränkel appliance produced no significant changes in maxillary growth during the evaluation period, while a statistically significant increase in mandibular length was observed. The maxillomandibular relationship improved mostly because of an increase in mandibular length. In addition, there were no statistically significant differences in the craniofacial growth direction between the Fränkel and the control group, both showing a slight downward rotation of the palatal plane. The Fränkel appliance produced a labial tipping of the lower incisors and a lingual inclination of the upper incisors as well as a significant increase in mandibular posterior dentoalveolar height. It was concluded that the main effects of the Fränkel appliance during this time period were mostly dentoalveolar with a smaller but significant skeletal mandibular effect.

Possible effects of sodium chloride treatment on quality of effluents from Alabama channel Catfish ponds

Channel catfish ponds are treated with salt (sodium chloride) to increase chloride concentration and prevent nitrite toxicity in fish. A survey indicated that most farmers try to maintain chloride concentration of 50 to 100 mg/L in ponds by annual salt applications. Averages and standard deviations for selected water quality variables in salt-treated ponds were as follows: chloride, 87.2 ± 37.5 mg/L; total dissolved solids (TDS), 336 ± 96 mg/L; specific conductance, 512 ± 164 μmhos/cm. Maximum values were 189 mg/L for chloride, 481 mg/L for TDS, and 825 μmhos/cm for specific conductance. Good correlations between specific conductance values and both chloride and TDS concentrations suggest that specific conductance can be a rapid method for estimating concentrations of these two variables in surface water. The maximum limit for chloride concentration in Alabama streams allowed by the Alabama Department of Environmental Management is 230 mg/L. The usual recommended upper limit of TDS for protection of aquatic life in freshwater streams is 1,000 mg/L. Based on the observed relationship between TDS concentration and specific conductance in Alabama catfish ponds, 1,000 mg/L TDS corresponds to 1,733 μmhos/cm specific conductance. It is unlikely that effluents from salt-treated catfish ponds would violate the in-stream chloride standard of 230 mg/L or harm aquatic life in streams. Nevertheless, chloride concentrations in ponds should be measured before salt application as a safe guard against excessive salt application and chloride concentrations above the in-stream chloride standard.

Net melon cultivars productivity under different cultivation systems, during summer and winter

In order to access the effects of cultivation in soil, sand or a commercial substrate on the productivity of Bonus #2 cultivars, 'Don Carlos' and 'Hy Mark', two experiments were conducted in the greenhouse at FCAV-UNESP, Jaboticabal- SP, Brazil, 21° 15' 22 S, 48° 18'58 W, and altitude of 595 m, from November 1999 to April 2000 (summer), and from July to November 2000 (Winter). For cultivation in soil, chemical nutrients were added, and plants were irrigated with drip irrigation. Fertigation with recirculation of the nutrient solution was used on sand; slabs were used on commercial substrate with the fertigation with non circulating nutrient solution. Bonus #2 cultivar yielded the highest production of marketable fruit, but were later in production, while Hy Mark cultivar had early production but a lower number of fruit per plant. The winter planting yielded higher production of marketable fruit while summer plantings yielded lower number of fruit per plant, but with higher average weigh. The combination between cultivation systems and cultivars yielded higher fruit production in winter.

Water table effects on bean yield and nitrate distribution in the soil profile

In order to evaluate the bean yield under different water table levels as well as the moisture and nitrate distribution in the soil profile, a field experiment was carried out at the experimental area from the College of Agronomic Sciences - UNESP, Botucatu, SP, Brazil. Beans were grown in field lysimeters and subjected to five water table depths:30; 40; 50; 60 and 70 cm. The moisture in the soil profile was gravimetrically determined through samples obtained at 10; 20; 30; 40; 50; 60 and 70cm of depth. The water table depths of 30cm and 40cm showed the highest productivities (3,228.4 kg.ha-1 and 3,422.1 kg.ha-1, respectively), showing no statistical differences between each other. The highest productivity was related to the two most elevated water table levels (30 and 40cm), which provided the highest moisture average values on basis of volume in the soil profile (33.3 e 31%) as well as the consumptive use of water (416 and 396 mm). The nitrate content during the bean cycle at the extraction depth of 60cm has been under the safe drinking limit of 10 mg.1-1 for water table depths of 30; 40; 50 and 60cm, showing the denitrification effectiveness as a way of controlling water table from nitrate pollution. The water table handling allowed the attainment of high bean productivity levels, as well as the reduction of the nitrate level.