Genetic and physiological alterations occurring in a yeast population continuously propagated at increasing temperatures with cell recycling

This work investigated the effects of increasing temperature from 30 degrees C to 47 degrees C on the physiological and genetic characteristics of Saccharomyces cerevisiae strain 63M after continuous fermentation with cell recycling in a system of five reactors in series. Steady state was attained at 30 degrees C, and then the temperature of the system was raised so it ranged from 35 degrees C in the last reactor to 43 degrees C in the first reactor or feeding reactor with a 2 degrees C difference between reactors. After 15 days at steady state, the temperature was raised from 37 degrees C to 45 degrees C for 25 days at steady state, then from 39 degrees C to 47 degrees C for 20 days at steady state. Starter strain 63M was a hybrid strain constructed to have a MAT a/alpha, LYS/lys, URA/ura genotype. This hybrid yeast showed vigorous growth on plates at 40 degrees C, weak growth at 41 degrees C, positive assimilation of melibiose, positive fermentation of galactose, raffinose and sucrose. of 156 isolates obtained from this system at the end of the fermentation process, only 17.3% showed the same characteristics as starter strain 63M. Alterations in mating type reaction and in utilization of raffinose, melibiose, and sucrose were identified. Only 1.9% of the isolates lost the ability to grow at 40 degrees C. Isolates showing requirements for lysine and uracil were also obtained. In addition, cell survival was observed at 39-47 degrees C, but no isolates showing growth above 41 degrees C were obtained.

Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp

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

Metabolic, ventilatory, and hygric physiology of a South American marsupial, the long-furred woolly mouse opossum

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cardioplegia using low volumic cardioplegic agents: morphological study in isolated rabbit hearts

INTRODUÇÃO: A solução cardioplégica farmacológica busca eliminar as conseqüências do dano isquêmico, que é o resultado do desbalanço entre a oferta e o consumo de energia durante a parada dos batimentos cardíacos, nas cirurgias cardíacas com circulação extracorpórea. OBJETIVO: Este trabalho avalia experimentalmente as alterações estruturais e ultra-estruturais em coração isolado de coelhos submetidos à parada protegida pela Solução para Cardioplegia de Baixo Volume (SCBV). MÉTODO: O estudo compreendeu um grupo controle e dois grupos experimentais. No grupo I, a parada cardíaca foi obtida pela infusão da SCBV por 2 horas. No grupo II, o experimento foi conduzido da mesma forma até a parada protegida pela SCBV por duas horas, imediatamente procedeu-se à reperfusão com solução oxigenada de Ringer Locke (RL) por uma hora. No grupo controle os corações foram perfundidos com solução oxigenada de RL por duas horas. Após os experimentos, oito amostras de parede lateral do ventrículo esquerdo foram fixadas em formaldeído 10% e glutaraldeído 2,5% para análises histológica e ultra-estrutural. RESULTADOS: As células do miocárdio, fibroblastos e células endoteliais, observadas nos grupos experimentais I e II, apresentaram marginalização da heterocromatina, compactação do nucléolo, alteração na forma das mitocôndrias, compactação das cristas e aumento da densidade da matriz mitocondrial, indicando que tanto a estrutura nuclear quanto a das organelas citoplasmáticas foram alteradas em relação às células do grupo controle. CONCLUSÃO: As modificações estruturais foram decorrentes de uma adaptação fisiológica da célula, não sendo indicativas de oncose ou apoptose, sugerindo, portanto, que a solução cardioplégica utilizada foi eficiente para a preservação das células.

Aptidão aeróbia e amplitude dos domínios de intensidade de exercício no ciclismo

INTRODUÇÃO: A determinação dos domínios de intensidade de exercício tem importantes implicações na prescrição do treino aeróbio e na elaboração de delineamentos experimentais. OBJETIVO: Analisar os efeitos do nível de aptidão aeróbia sobre a amplitude dos domínios de intensidade de exercício durante o ciclismo. MÉTODOS: Doze ciclistas (CIC), 11 corredores (COR) e oito indivíduos não treinados (NT) foram submetidos aos seguintes protocolos em diferentes dias: 1) teste progressivo para determinação do limiar de lactato (LL), consumo máximo de oxigênio (VO2máx) e sua respectiva intensidade (IVO2máx); 2) três testes de carga constante até a exaustão a 95, 100 e 110% IVO2máx para a determinação da potência crítica (PC); 3) testes até a exaustão para determinar a intensidade superior do domínio severo (Isup). As amplitudes dos domínios (moderado < LL; LL > pesado < PC; PC > severo < Isup) foram expressas como percentual da Isup (VO2). RESULTADOS: A amplitude do domínio moderado foi similar entre CIC (52 ± 8%) e COR (47 ± 4%) e significantemente maior no CIC em relação ao NT (41 ± 7%). O domínio pesado foi significantemente menor no CIC (17 ± 6%) em relação ao COR (27 ± 6%) e NT (27 ± 9%). Em relação ao domínio severo não foram encontradas diferenças significantes entre os CIC (31 ± 7%), COR (26 ± 5%) e NT (31 ± 7%). CONCLUSÃO: O domínio pesado de exercício é mais sensível a mudanças determinadas pelo nível de aptidão aeróbia, existindo a necessidade de que se atenda ao princípio da especificidade do movimento, quando se pretende obter um elevado grau de adaptação fisiológica.

Effect of thermal conditioning during embryonic development on aspects of physiological responses of broilers to heat stress

The purpose of the present study was to investigate the effect of thermal conditioning, (through exposure to heat stress), during pre-hatch development on some physiological responses of post-hatch broilers to a post-natal heat stress challenge. Exposure to heat stress at this stage, we hope, may possibly induce epigenetic heat adaptation. Incubating eggs were exposed to temperature of 39.0degreesC for 2 h from Day 13 to 17 of incubation. At 33, 35, 37, 39, 41 and 43 d of age, the broilers hatched from these eggs were housed individually in open-circuit respiration cells. The climatic chambers were set to 22degreesC and increased to 30degreesC for 4 h. O-2 consumption and CO2 production of each chicken was monitored continuously in order to calculate the heat production. Blood samples were obtained before and during the 4 h heat stress. Thermal conditioning during incubation did not affect the plasma T-4, corticosterone, glucose, uric acid and CK concentrations. Temperature challenge, decreased plasma T-3 of broilers of both groups but the decrease was greater in pre-conditioned broilers compared with controls. A similar trend was observed for triglycerides. These changes did not affect total heat production. Since decreased T3 and triglyceride levels are part of the mechanisms for thermoregulation, these suggest that thermal conditioning during incubation can improve the broiler chicken capability for thermotolerance at later post-hatch age. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

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.

Physiological responses of broiler chickens to heat stress and dietary electrolyte balance (sodium plus potassium minus chloride, milliequivalents per kilogram)

Individually caged male Cobb broilers (24), 44 d of age, were used to evaluate effects of heat stress (1 d of data collection) and dietary electrolyte balance (DEB; Na + K - Cl, mEq/kg from 1 d of age). During summer rearing, mortality was variable, but DEB 240 improved growth, feed conversion ratio, water intake, and waterrfeed ratio vs. DEB 0. The temperature sequence for heat stress was 24 to 32°C in 30 min, 32 to 36°C in 30 min, 36 to 37°C in 15 min, and 37 to 41°C in 45 min. Maximum temperature was held for 15, 60, 90, or 360 min for data collection (relative humidity averaged 42 ± 7%). Results from the same room before and after heat stress were analyzed by DEB (1-factor ANOVA) and before vs. after heat stress compared across DEB (2-sample t-test). Heat stress decreased blood Na, K, and pCO2, and lymphocytes but increased heterophils. Blood HCO3 rose, Cl declined, and hematocrit gave a concave pattern (lowest at DEB 120) as DEB increased. After heat stress, DEB O decreased blood Na and K, and DEB O and 120 levels decreased blood HCO3. After heat stress blood pCO2 and hemoglobin decreased with DEB 240, but it had highest pCO2, a key factor. The DEB 120 gave longest times to panting and prostration with DEB O and 240 results lower but similar statistically. In heat stress, DEB 360 was excessive, DEB 120 and 240 were favorable, and DEB 0 was intermediate based on hematology, panting, and prostration responses.

Maxi-K channels contribute to urinary potassium excretion in the ROMK-deficient mouse model of Type II Bartter's syndrome and in adaptation to a high-K diet

Type II Bartter's syndrome is a hereditary hypokalemic renal salt-wasting disorder caused by mutations in the ROMK channel (Kir1.1; Kcnj1), mediating potassium recycling in the thick ascending limb of Henle's loop (TAL) and potassium secretion in the distal tubule and cortical collecting duct (CCT). Newborns with Type II Bartter are transiently hyperkalemic, consistent with loss of ROMK channel function in potassium secretion in distal convoluted tubule and CCT. Yet, these infants rapidly develop persistent hypokalemia owing to increased renal potassium excretion mediated by unknown mechanisms. Here, we used free-flow micropuncture and stationary microperfusion of the late distal tubule to explore the mechanism of renal potassium wasting in the Romk-deficient, Type II Bartter's mouse. We show that potassium absorption in the loop of Henle is reduced in Romk-deficient mice and can account for a significant fraction of renal potassium loss. In addition, we show that iberiotoxin (IBTX)-sensitive, flow-stimulated maxi-K channels account for sustained potassium secretion in the late distal tubule, despite loss of ROMK function. IBTX-sensitive potassium secretion is also increased in high-potassium-adapted wild-type mice. Thus, renal potassium wasting in Type II Bartter is due to both reduced reabsorption in the TAL and K secretion by max-K channels in the late distal tubule. © 2006 International Society of Nephrology.

Monitoring chronic physical stress using biomarkers, performance protocols and mathematical functions to identify physiological adaptations in rats

This study was undertaken to characterize the effects of monotonous training at lactate minimum (LM) intensity on aerobic and anaerobic performances; glycogen concentrationsin the soleus muscle, the gastrocnemius muscle and the liver; and creatine kinase (CK), free fatty acids and glucose concentrations in rats. The rats were separated into trained (n =10), baseline (n = 10) and sedentary (n=10) groups. The trained group was submitted to the following: 60 min/day, 6 day/week and intensity equivalent to LM during the 12-week training period. The training volume was reduced after four weeks according to a sigmoid function. The total CK (U/L) increased in the trained group after 12 weeks (742.0±158.5) in comparison with the baseline (319.6±40.2) and the sedentary (261.6+42.2) groups. Free fatty acids and glycogen stores (liver, soleus muscle and gastrocnemius muscle) increased after 12 weeks of monotonous training but aerobic and anaerobic performances were unchanged in relation to the sedentary group. The monotonous training at LM increased the level of energy substrates, unchanged aerobic performance, reduced anaerobic capacity and increased the serum CK concentration; however, the rats did not achieve the predicted training volume.

Assessment of the physiological potential of spinach seeds (Tetragonia tetragonoides (Pall.) Kuntze)

The efficiency of vigor tests in assessing the physiological potential of seeds depends on their standardization for different species. In this context, the research aimed to study specific methodologies to evaluate the vigor of spinach seeds (Tetragonia tetragonoides). For this purpose, five lots of spinach seed cv. New Zealand were submitted to germination, first count of germination, seedling emergence in the field, accelerated aging (with and without use of saturated solution of NaCl at 41and 45 ºC for 24,48 and 72 hours), controlled deterioration (18,21 and 24% seed water content adjustments for 24 hours at 45 ºC) and electrical conductivity tests with variations inwater volume (25, 50 and 75 mL), seed quantity (25 and 50) and soaking period (1, 2, 4, 8,12, 16 and 24 hours) at 25 ºC. The first count test allowed obtaining preliminary information about seed vigor, and other tests in general showed similarity with seedling emergence in the field. Thus, it was concluded that traditional accelerated aging test 24 h /41 ºC and accelerated aging test with saturated salt solution for 24 h at 41 ºC and 45 ºC, and controlled deterioration test with 21% seed water content / 24 h / 45 ºC were all efficient for evaluating the physiological potential of spinach seeds.

Wolf presence in the ranch of origin: Impacts on temperament and physiological responses of beef cattle following a simulated wolf encounter

This experiment evaluated temperament, vaginal temperature, and plasma cortisol in beef cows from wolf-naive and wolf-experienced origins that were subjected to a simulated wolf encounter. Multiparous, pregnant, nonlactating Angus-crossbreed cows from the Eastern Oregon Agricultural Research Center located near Burns, OR (CON; n = 50), and from a commercial operation near Council, ID (WLF; n = 50), were used. To date, grey wolves are not present around Burns, OR, and thus CON were naive to wolves. Conversely, wolves are present around Council, ID, and WLF cows were selected from a herd that had experienced multiple confirmed wolf-predation episodes from 2008 to 2012. Following a 50-d commingling and adaptation period, CON and WLF cows were ranked by temperament, BW, and BCS and allocated to 5 groups (d 0; 10 CON and 10 WLF cows/group). Groups were individually subjected to the experimental procedures on d 2 (n = 3) and d 3 (n = 2). Before the simulated wolf encounter, cow temperament was assessed and blood samples and vaginal temperatures (using intravaginal data loggers) were collected (presimulation assessments). Cows were then sorted by origin, moved to 2 adjacent drylot pens (10 WLF and 10 CON cows/pen), and subjected to a simulated wolf encounter event for 20 min, which consisted of 1) cotton plugs saturated with wolf urine attached to the drylot fence, 2) continuous reproduction of wolf howls, and 3) 3 leashed dogs that were walked along the fence perimeter. Thereafter, WLF and CON cows were commingled and returned to the handling facility for postsimulation assessments, which were conducted immediately after exposure to wolf-urine-saturated cotton plugs, wolf howl reproduction, and 20-s exposure to the 3 dogs while being restrained in a squeeze chute. Chute score, temperament score, and plasma cortisol concentration increased (P <= 0.01) from pre- to postsimulation assessment in WLF but did not change in CON cows (P >= 0.19). Exit velocity decreased (P = 0.01) from pre-to postsimulation assessment in CON but did not change (P = 0.79) in WLF cows. In addition, WLF cows had a greater (P = 0.03) increase in temperature from pre-to postsimulation assessments compared with CON cows. In conclusion, the simulated wolf encounter increased excitability and fear-related physiological stress responses in cows that originated from a wolf-experienced herd but not in cows that originated from a wolf-naive herd.