POTASSIUM FERTILIZATION, ROOT MORPHOLOGY AND POTASSIUM ABSORPTION BY SOYBEAN

A greenhouse experiment studied the effect of potassium fertilization on soybean (Glycine max L. Merrill) root morphology and on K absorption by six soybean cultivars of different maturation groups and growth habits. The Plants were grown up to 70 days after plant emergence, in pots containing 6.0 kg of soil. In the absence of K, no significant difference in K absorption was observed among the cultivars or in root length and surface, but root mean radius was correlated to K absorption. Differences in K absorption were not associated with root characteristics in the presence of K fertilization. Physiological adjustments in K uptake, as well as K availability in the soil, were more important in soybean nutrition than were morphological adjustments in the root system. The results were not associated with plant growth habit or with maturation group.

Role of roscovitine and IBMX on kinetics of nuclear and cytoplasmic maturation of bovine oocytes in vitro

The 3-isobutyl-1-methylxanthine (IBMX) is able to prevent resumption of meiosis by maintaining elevated cyclic AMP (cAMP) concentrations in the oocyte, and roscovitine, a purine known to specifically inhibit MPF kinase activity, maintains bovine oocytes at the germinal vesicle (GV) stage. The present study was conducted to analyze whether cytoplasmic maturation (examined by the pattern of cortical granule (CG) distribution) of bovine oocytes is improved during meiotic arrest with IBMX and roscovitine. Oocytes were matured in vitro in a 10% Knockout(SR) supplemented TCM-199 medium (Control) with either 0.5 mM IBMX or 25 mu M roscovitine (ROSC). Oocytes were stained with fluorescein isothiocyanate conjugated Lens culinaris agglutinin (FITC-LCA) for CG evaluation and with Hoechst 33342 for nuclear stage assessment. At 16 h of culture, the percentage of oocytes remaining in the GV stage was higher (P < 0.05) in the ROSC group (32.41%) compared with the Control and IBMX groups (8.61% and 9.73%, respectively). At 24h of culture, progression of meiosis to M II stage was retarded (P < 0.05) in the ROSC group (24.05%) compared to the Control (60.20%), whereas the IBMX group (33.88%) showed no significant difference to the other two groups. At 16h of maturation, the proportion of oocytes with CG in clusters (immature cytoplasm) was similar between the groups, as was the percentage of peripheral CG (mature) at 24h of maturation. The results of the present study demonstrated that the meiotic inhibitors IBMX and roscovitine delay the progression of nuclear maturation without affecting cytoplasmic maturation, assessed by the analysis of CG repositioning. (c) 2006 Elsevier B.V. All rights reserved.

Effect of the chronological age and sexual maturation on the time to exhaustion at maximal aerobic speed

The purposes of this study were: a) to verify the effect of chronological age and sexual maturation on the time to exhaustion at VO(2)max (t(lim)) and; b) to examine the reproducibility of t(lim) in boys aged 10-15 years. Forty boys, divided into 4 groups, in accordance to the chronological age (G10-12 and G13-15) and sexual maturation (P1-P3 and P4-P5 levels for pubic hair), performed the following tests: 1) incremental test for determination of VO(2)max and; 2) all-out exercise bout performed at VO(2)max to determine the t(lim). There was no difference of t(lim) (sec) between G10-12 and G13-15 (181.5 +/- 96.3 vs. 199 105.5). While the two measures of t(lim) were moderately related (r = 0.78), t(lim) from the second test (226.6 +/- 96.1 s) was higher than that of the first (191.3 +/- 79.2 s). We can conclude that the t(lim) is not influenced by chronological age and sexual maturation. Besides, t(lim) presents a lower reproducibility in children and adolescents.

THE EFFECT OF SERUM ON IN-VITRO MATURATION, IN-VITRO FERTILIZATION AND STEROIDOGENESIS OF BOVINE OOCYTES COCULTURED WITH GRANULOSA-CELLS

The influence of fetal calf serum alone (FCS) or associated with proestrous (FCS+PCS), estrous (FCS+ECS) or metaestrous (FCS+MCS) cow serum added to the culture medium and of the steroids produced by co-cultured granulosa cells were evaluated in terms of the in vitro maturation (TVM) and fertilization (IVF) of bovine oocytes. Supplementation of the medium with FCS+ECS and FCS+MCS resulted in higher proportions of oocytes that reached metaphase II (96.0% and 93.3%, respectively) and in higher proportions of embryos that reached the four- and eight-cell/morula stages (51.9% and 65.6%, respectively), whereas the supplementation with FCS and FCS+PCS resulted in only 79.2% and 67.5%, respectively, of matured oocytes and 26.7% and 34.3%, respectively, of cleaved embryos. These findings show that the best IVM and IVF were obtained at lower concentrations of estradiol produced by co-cultured granulosa cells (supplementation with FCS+ECS: 10.3 ng/ml and FCS+MCS: 2.1 ng/ml), whereas the worst-results in IVM and IVF occurred at higher concentrations of estradiol that were obtained with FCS (33.1 ng/ml) and FCS+PCS (19.9 ng/ml) supplementation. These data suggest an inhibitory effect of estradiol on resumption of oocyte meiosis in vitro.

The effect of timing of thermal conditioning during incubation on embryo physiological parameters and its relationship to thermotolerance in adult broiler chickens

Previously, we reported that thermal conditioning at 39degreesC on days 13-17 of incubation of broiler eggs enabled thermotolerance during post-hatch growth (J. Therm. Biol. 28 (2003) 133). Tolerance to a temperature of 30degreesC was accompanied by changes in thyroid hormones and metabolic parameters. In the current study, we determined the mechanism of epigenetic heat adaptation during embryonic age by measuring blood physiological parameters that may be associated with the ultimate effects of thermal conditioning. Hatching eggs from Ross breeders were subjected to heat treatment of 39degreesC at days 13, 14, 15, 16 and 17 of incubation for 2 h per day. Control eggs were incubated at 37.6degreesC. Samples of eggs were withdrawn on each day of thermal conditioning and at internal pipping (IP) to obtain blood samples from embryos. The remaining eggs were weighed at day 18 and transferred to hatchers. The timing of IP, external pipping (EP) and hatching were monitored every 2 h. At hatch, chicks were weighed and hatchability was determined. Blood samples were obtained from samples of day-old chicks. T3, T4, corticosterone, pCO(2), pO(2) levels were determined in the blood. Blood pH was measured and T3/T4 ratios were calculated. Heat conditioning significantly increased corticosterone and pO(2) levels and blood pH but depressed pCO(2) at day 14. These were followed by a significant depression of T4 level on day 15. Remarkably, at day 16, all these parameters were back to normal as in the control embryos. Hatching was delayed by thermal conditioning probably as a result of the depressed corticosterone levels at IP. Hatchability was also lower in the heat-treated group but 1-day old chick weights were comparable to those of the controls. The result suggests that epigenetic thermal conditioning involves changes in these physiological parameters and probably serve as a method for epigenetic temperature adaptation since the same mechanisms are employed for coping with heat during post-embryonic growth. It also suggests that days 14-15 may be the optimal and most sensitive timing for evoking this mechanism during embryonic development. The adverse effects of heat treatment observed in this study may have been due to the continued exposure to heat until day 17. Fine-tuning thermal conditioning to days 14-15 only may improve these production parameters. (C) 2003 Elsevier Ltd. All rights reserved.

Kinetic characterization of hypophosphatasia mutations with physiological substrates

We have analyzed 16 missense mutations of the tissue-nonspecific AP (TNAP) gene found in patients with hypophosphatasia. These mutations span the phenotypic spectrum of the disease, from the lethal perinatal/infantile forms to the less severe adult and odontohypophosphatasia. Site-directed mutagenesis was used to introduce a sequence tag into the TNAP cDNA and eliminate the glycosylphosphatidylinositol (GPI)-anchor recognition sequence to produce a secreted epitope-tagged TNAP (setTNAP). The properties of GPI-anchored TNAP (gpiTNAP) and setTNAP were found comparable. After introducing each single hypophosphatasia mutation, the setTNAP and mutant TNAP cDNAs were expressed in COS-1 cells and the recombinant flagged enzymes were affinity purified. We characterized the kinetic behavior, inhibition, and heat stability properties of each mutant using the artificial substrate p-nitrophenylphosphate (pNPP) at pH 9.8. We also determined the ability of the mutants to metabolize two natural substrates of TNAP, that is, pyridoxal-5'-phosphate (PLP) and inorganic pyrophosphate (PPi), at physiological pH. Six of the mutant enzymes were completely devoid of catalytic activity (R54C, R54P, A94T, R206W, G317D, and V365I), and 10 others (A16V, A115V, A160T, A162T, E174K, E174G, D277A, E281K, D361V, and G439R) showed various levels of residual activity. The A160T substitution was found to decrease the catalytic efficiency of the mutant enzyme toward pNPP to retain normal activity toward PPi and to display increased activity toward PLP. The A162T substitution caused a considerable reduction in the pNPPase, PPiase, and PLPase activities of the mutant enzyme. The D277A mutant was found to maintain high catalytic efficiency toward pNPP as substrate but not against PLP or PPi. Three mutations ( E174G, E174K, and E281K) were found to retain normal or slightly subnormal catalytic efficiency toward pNPP and PPi but not against PLP. Because abnormalities in PLP metabolism have been shown to cause epileptic seizures in mice null for the TNAP gene, these kinetic data help explain the variable expressivity of epileptic seizures in hypophosphatasia patients.

PROLACTIN INDUCES MATURATION OF GLUCOSE SENSING MECHANISMS IN CULTURED NEONATAL RAT ISLETS

The effects of PRL treatment on insulin content and secretion, and Rb-86 and Ca-45 fluxes from neonatal rat islets maintained in culture for 7-9 days were studied. PRL treatment enhanced islet insulin content by 40% and enhanced early insulin secretion evoked by 16.7 mm glucose. Insulin release stimulated by oxotremorine-M, a muscarinic agonist, in the presence of glucose (8.3 or 16.7 mm) was unchanged by PRL treatment. However, PRL treatment potentiated phorbol 12,13-dibutyrate-stimulated insulin secretion in the presence of the above glucose concentrations. PRL treatment potentiated the reduction in Rb-86 efflux induced by glucose or tolbutamide and enhanced the increase in Rb-86 efflux evoked by diazoxide. PRL treatment slightly potentiated the increment in Ca-45 uptake induced by high concentrations of K+, but failed to affect the increment evoked by 16.7 mm glucose. Since glucose-induced Ca-45 uptake was not affected by PRL, we suggest that the enhancement in first phase insulin secretion evoked by glucose in the PRL-treated islets occurs at a step in the secretory process that may involve protein kinase-C. These data further support observations that PRL treatment increases islet sensitivity to glucose.

Interval training at 95% and 100% of the velocity at VO2 max: effects on aerobic physiological indexes and running performance

The objective of this study was to analyze the effect of two different high-intensity interval training (HIT) programs on selected aerobic physiological indices and 1500 and 5000 m running performance in well-trained runners. The following tests were completed (n = 17): (i) incremental treadmill test to determine maximal oxygen uptake (VO2max), running velocity associated with VO2 max (VVO2max), and the velocity corresponding to 3.5 mmol/L of blood lactate concentration (vOBLA); (ii) submaximal constant-intensity test to determine running economy (RE); and (iii) 1500 and 5000 m time trials on a 400 m track. Runners were then randomized into 95% vVO(2max) or 100% vVO(2max) groups, and undertook a 4 week training program consisting of 2 HIT sessions (performed at 95% or 100% vVO(2max), respectively) and 4 submaximal run sessions per week. Runners were retested on all parameters at the completion of the training program. The VO2 max values were not different after training for both groups. There was a significant increase in post-training vVO(2 max), RE, and 1500 in running performance in the 100% vVO(2 max) group. The vOBLA and 5000 m running performance were significantly higher after the training period for both groups. We conclude that vOBLA and 5000 m running performance can be significantly improved in well-trained runners using a 4 week training program consisting of 2 HIT sessions (performed at 95% or 100% vVO(2max)) and 4 submaximal run sessions per week. However, the improvement in vVO(2 max), RE, and 1500 in running performance seems to be dependent on the HIT program at 100% vVO(2 max).

Seed maturation and effect of temperature regime on Trema micrantha (L.) Blume seed germination

T. micrantha (L.) Blume (Ulmaceae), a common pioneer tree species in Brazil, is used in the restoration of degraded areas. The fruits are fleshy and indehiscent, with only one water impermeable seed. During the fruiting period, fruits of different colours are found at the same time on the same branch. This research aimed to correlate fruit colour with other physical indicators of seed maturity and to Verify the effect of temperature regime on seed germination. Collected fruits were separated in to green, green-red and red colour and for each of these maturation stages, size, moisture content and dry matter of both fruits and seeds were determined. Seeds were scarified with sulphuric acid and submitted to a germination test conducted at constant (20 degreesC, 30 degreesC and 40 degreesC) and alternating (20-30 degreesC, 30-40 degreesC and 20-40 degreesC) temperatures for 15 weeks. Seed germination percentage and speed were analysed after five weeks and the fmal percentage of germinated and Viable seeds after 15 weeks. Seed maturity is attained when the fruits are green-red. At this stage, moisture content was about 64% for fruits and 10% for seeds. Alternating temperature was required for seed germination and 20-30 degreesC was the best option. Most seeds had germinated after five weeks, providing mature seeds, acid scarification and alternating temperature were used.

Synthesis and characterization of phosphorylcholine-substituted chitosans soluble in physiological pH conditions

A polymer analogous synthesis involving the reductive amination of phosphorylcholine (PC)-glyceraldehyde with primary amines of deacetylated chitosan (M-w approximate to 57000 g mol(-1)) was used to prepare phosphorylcholine-substituted chitosans (PC-CH) with a degree of substitution (DS) ranging from similar to 11 to similar to 53 mol% PC-substituted glucosamine residues. The PC-CH derivatives were characterized by H-1 NMR spectroscopy, FTIR spectroscopy, and multiangle laser light scattering gel permeation chromatography (MALLS-GPC). The pKa of the PC-substituted amine groups (pKa approximate to 7.20) was determined by H-1 NMR titration. The PC-CH samples (1.0 g L-1) were shown to be nontoxic using an MTT assay performed with human KB cells. Aqueous solutions of PC-CH samples (4.0 g L-(1)) of DS g 22 mol% PC-substituted glucosamine residues remained clear, independently of pH (4.0 < pH < 11.0). The remarkable water solubility and nontoxicity displayed by the new PC-CH samples open up new opportunities in the design of chitosan-based biomaterials and nanoparticles.

Effect of thermal stress on physiological parameters, feed intake and plasma thyroid hormones concentration in Alentejana, Mertolenga, Frisian and Limousine cattle breeds

The aim of the present study was to assess the heat tolerance of animals of two Portuguese (Alentejana and Mertolenga) and two exotic (Frisian and Limousine) cattle breeds, through the monitoring of physiological acclimatization reactions in different thermal situations characterized by alternate periods of thermoneutrality and heat stress simulated in climatic chambers. In the experiment, six heifers of the Alentejana, Frisian and Mertolenga breeds and four heifers of the Limousine breed were used. The increase in chamber temperatures had different consequences on the animals of each breed. When submitted to heat stress, the Frisian animals developed high thermal polypnea (more than 105 breath movements per minute), which did not prevent an increase in the rectal temperature (from 38.7 degrees C to 40.0 degrees C). However, only a slight depression in food intake and in blood thyroid hormone concentrations was observed under thermal stressful conditions. Under the thermal stressful conditions, Limousine animals decreased food intake by 11.4% and blood triiodothyronine (T3) hormone concentration decreased to 76% of the level observed in thermoneutral conditions. Alentejana animals had similar reactions. The Mertolenga cattle exhibited the highest capacity for maintaining homeothermy: under heat stressful conditions, the mean thermal polypnea increased twofold, but mean rectal temperature did not increase. Mean food intake decreased by only 2% and mean T3 blood concentration was lowered to 85,6% of the concentration observed under thermoneutral conditions. These results lead to the conclusion that the Frisian animals had more difficulty in tolerating high temperatures, the Limousine and Alentejana ones had an intermediate difficulty, and the Mertolenga animals were by far the most heat tolerant.

EVALUATION of COVERING MATERIALS IN INDIVIDUAL SHELTERS and ITS EFFECTS on PHYSIOLOGICAL RESPONSES and PERFORMANCE of DAIRY CALVES

In Tropical regions, the animal performance is often affected by climate conditions. This study aimed to evaluate covering materials in individual shelters, normally used to house dairy calves, and its influence on the calves physiology and performance. The design used was completely randomized, with a 2x3 factorial arrangement to compare the averages of 5% through the Tukey's test, i.e., both genders-and three types of covering in the shelters (Z - zinc; AC - asbestos cement; and WPAC - white-painted asbestos cement). Parameters evaluated included daily weight gain (DWG), dry matter intake (DMI), feed conversion (FC), rectal temperature (RT), and respiratory frequency (RF). Results showed significant differences (P < 0.05) among males (1.04kg/day) and females (0.74kg/day) for DWG and interaction between gender and treatment (P < 0.05) for zinc covering (0.562kg/day for females and 1.120kg/day for males). Significant differences were also observed in FI of animals housed under shelters with the covering of zinc (48.35kgDM/day for females and 96.91 kgDM/day for males). There were no significant differences (P > 0.05) in the FC and the RT, and there were significant differences (P < 0.05) for RF in the Z treatments (56.9 mov.min(-1)), WPAC (62.2 mov.min(-1)) and FC (70.25 mov.min-1). It was concluded that different covering materials did not affect performance and dry matter intake of dairy calves. However, the animals' physiology of thermoregulation was altered by the different covering materials used in individual shelters.