Protein dynamics: hydration and cavities

The temperature-pressure behavior of proteins seems to be unique among the biological macromolecules. Thermodynamic as well as kinetic data show the typical elliptical stability diagram. This may be extended by assuming that the unfolded state gives rise to volume and enthalpy-driven liquid-liquid transitions. A molecular interpretation follows from the temperature and the pressure dependence of the hydration and cavities. We suggest that positron annihilation spectroscopy can provide additional quantitative evidence for the contributions of cavities to the dynamics of proteins. Only mature amyloid fibrils that form from unfolded proteins are very resistant to pressure treatment.

Modeling the pressure inactivation dynamics of Escherichia coli

Escherichia coli, as a model microorganism, was treated in phosphate-buffered saline under high hydrostatic pressure between 100 and 300 MPa, and the inactivation dynamics was investigated from the viewpoint of predictive microbiology. Inactivation data were curve fitted by typical predictive models: logistic, Gompertz and Weibull functions. Weibull function described the inactivation curve the best. Two parameters of Weibull function were calculated for each holding pressure and their dependence on holding pressure was obtained by interpolation. With the interpolated parameters, inactivation curves were simulated and compared with the experimental data sets.

Influence of He-Ne laser therapy on the dynamics of wound healing in mice treated with anti-inflammatory drugs

We determined the effects of helium-neon (He-Ne) laser irradiation on wound healing dynamics in mice treated with steroidal and non-steroidal anti-inflammatory agents. Male albino mice, 28-32 g, were randomized into 6 groups of 6 animals each: control (C), He-Ne laser (L), dexamethasone (D), D + L, celecoxib (X), and X + L. D and X were injected im at doses of 5 and 22 mg/kg, respectively, 24 h before the experiment. A 1-cm long surgical wound was made with a scalpel on the abdomens of the mice. Animals from groups L, D + L and X + L were exposed to 4 J (cm²)-1 day-1 of He-Ne laser for 12 s and were sacrificed on days 1, 2, or 3 after the procedure, when skin samples were taken for histological examination. A significant increase of collagen synthesis was observed in group L compared with C (168 ± 20 vs 63 ± 8 mm²). The basal cellularity values on day 1 were: C = 763 ± 47, L = 1116 ± 85, D = 376 ± 24, D + L = 698 ± 31, X = 453 ± 29, X + L = 639 ± 32 U/mm². These data show that application of L increases while D and X decrease the inflammatory cellularity compared with C. They also show that L restores the diminished cellularity induced by the anti-inflammatory drugs. We suggest that He-Ne laser promotes collagen formation and restores the baseline cellularity after pharmacological inhibition, indicating new perspectives for laser therapy aiming to increase the healing process when anti-inflammatory drugs are used.

Dynamics of immunosuppression in hamsters with experimental visceral leishmaniasis

Immunosuppression has been reported to occur during active visceral leishmaniasis and some factors such as the cytokine profile may be involved in this process. In the mouse model of cutaneous leishmaniasis using Leishmania (Leishmania) major, the Th1 response is related to protection while the Th2 response is related to disease progression. However, in hamsters, which are considered to be an excellent model for the study of visceral leishmaniasis, this dichotomy is not observed. Using outbred 45- to 60-day-old (140 to 150 g) male hamsters infected intraperitoneally with 2 x 10(7) L. (L.) chagasi amastigotes, we evaluated the immune response of spleen cells and the production of cytokines. We used 3 to 7 hamsters per group evaluated. We detected a preserved response to concanavalin A measured by index of proliferation during all periods of infection studied, while a proliferative response to Leishmania antigen was detected only at 48 and 72 h post-infection. Messenger RNA from cytokines type 1 (IL-2, TNF-α, IFN-γ) and type 2 (IL-4, IL-10 and TGF-β) detected by reverse transcriptase polymerase chain reaction and produced by spleen cells showed no qualitative difference between control non-infected hamsters and infected hamsters during any period of infection evaluated. Cytokines were measured by the DNA band intensity on agarose gel using the Image Lab 1D L340 software with no differences observed. In conclusion, the present results showed an antigen-dependent immunosuppression in hamsters with active visceral leishmaniasis that was not related to the cytokine profile.

Dynamics of chest wall volume regulation during constant work rate exercise in patients with chronic obstructive pulmonary disease

This study evaluated the dynamic behavior of total and compartmental chest wall volumes [(V CW) = rib cage (V RC) + abdomen (V AB)] as measured breath-by-breath by optoelectronic plethysmography during constant-load exercise in patients with stable chronic obstructive pulmonary disease. Thirty males (GOLD stages II-III) underwent a cardiopulmonary exercise test to the limit of tolerance (Tlim) at 75% of peak work rate on an electronically braked cycle ergometer. Exercise-induced dynamic hyperinflation was considered to be present when end-expiratory (EE) V CW increased in relation to resting values. There was a noticeable heterogeneity in the patterns of V CW regulation as EEV CW increased non-linearly in 17/30 "hyperinflators" and decreased in 13/30 "non-hyperinflators" (P < 0.05). EEV AB decreased slightly in 8 of the "hyperinflators", thereby reducing and slowing the rate of increase in end-inspiratory (EI) V CW (P < 0.05). In contrast, decreases in EEV CW in the "non-hyperinflators" were due to the combination of stable EEV RC with marked reductions in EEV AB. These patients showed lower EIV CW and end-exercise dyspnea scores but longer Tlim than their counterparts (P < 0.05). Dyspnea increased and Tlim decreased non-linearly with a faster rate of increase in EIV CW regardless of the presence or absence of dynamic hyperinflation (P < 0.001). However, no significant between-group differences were observed in metabolic, pulmonary gas exchange and cardiovascular responses to exercise. Chest wall volumes are continuously regulated during exercise in order to postpone (or even avoid) their migration to higher operating volumes in patients with COPD, a dynamic process that is strongly dependent on the behavior of the abdominal compartment.

Effect of chickpea (Cicer arietinum L.) germination on the major globulin content and in vitro digestibility

Chickpea seed germination was carried out over a period of 6 days. Little variation in the nitrogen and total globulin content was observed. The major globulin (11 S type) showed higher variation after the 4th day of germination. The elution behaviour and distribution of the isolated major globulin fraction on Sepharose CL-6B chromatography showed little modification at the end of germination. On SDS-PAGE the peak eluted from Sepharose CL-6B showed changes in protein bands between 20 and 30 kDa and above 60 kDa, indicating protein degradation during the period. Proteolytic activity was detected in the albumin fraction of the seeds, which increased up to the fourth and then decreased up to the sixth day, when isolated chickpea total globulin and casein were used as substrates. Chickpea flour, isolated albumin and total globulin fractions did not show an increase for in vitro digestibility; however, the isolated major globulin was more susceptible to hydrolysis after germination.

Using Computational Fluid-Dynamics (CFD) for the evaluation of beer pasteurization: effect of orientation of cans

In-package pasteurization is the most used method for beer microbiological stabilization. The search for safer and better quality food has created a need to better understand the processes involved in producing it. However, little is known about the temperature and velocity profiles during the thermal processes of liquid foods in commercial packaging, which results in over-dimensioned processes to guarantee safety, decreasing the sensorial and nutritional characteristics of the product and increasing process costs. Simulations using Computational Fluid-Dynamics (CFD) have been used by various authors to evaluate those processes. The objective of the present paper was to evaluate the effect of packaging orientation in the pasteurization of beer in a commercial aluminum can using CFD. A heating process was simulated at 60 ºC up to 15 PUs (a conventional beer process, in which 1 Pasteurization Unit (PU) is equivalent to 1minute at 60 ºC). The temperature profile and convection current velocity along the process and the variation of the PUs were evaluated in relation to time considering the cans in the conventional, inverted, and horizontal positions. The temperature and velocity profiles were similar to those presented in the literature. The package position did not result in process improvement.

Computational fluid dynamics evaluation of liquid food thermal process in a brick shaped package

Food processes must ensure safety and high-quality products for a growing demand consumer creating the need for better knowledge of its unit operations. The Computational Fluid Dynamics (CFD) has been widely used for better understanding the food thermal processes, and it is one of the safest and most frequently used methods for food preservation. However, there is no single study in the literature describing thermal process of liquid foods in a brick shaped package. The present study evaluated such process and the influence of its orientation on the process lethality. It demonstrated the potential of using CFD to evaluate thermal processes of liquid foods and the importance of rheological characterization and convection in thermal processing of liquid foods. It also showed that packaging orientation does not result in different sterilization values during thermal process of the evaluated fluids in the brick shaped package.

Dynamics of gas levels inside packages containing minimally processed Pera orange

The purpose of this study was to evaluate the changes in concentrations of O2 and CO2 inside packages of minimally processed Pera orange. Previously selected oranges that were washed, sanitized, and chilled were peeled using hydrothermal treatment (immersion of fruits in water at 50 °C for 8 minutes). The peeled oranges were then packed in five different plastic packages under passive and active modified atmosphere (5% O2 + 10% CO2 + 85% N2). The fruits were stored at 6 °C and 12 °C. The package headspace gas composition was evaluated for twelve days at 6 °C and nine days at 12 °C. The polypropylene film (32 µm) promoted modified atmosphere similar to that initially injected (5% O2 + 10% CO2 + 85% N2) at 6 °C and 12 °C. With regard to the atmosphere modification system, the injection of a gas mixture anticipated achieving an equilibrium atmosphere inside the packages at 12 °C. At 6 °C, the gas composition inside the packages was kept close to that of the injection, but the equilibrium was not verified.

Heliotropism and water availability effects on flowering dynamics and seed production in Macroptilium lathyroides

The experiment was carried out in pots in a glasshouse, with one plant per pot and nine repetitions per treatment. The treatments consisted of free or restricted leaves, submited to 90-100% or 60-70% soil field capacity (FC). Only independent effects of water availability or leaf movement were observed on yield components. Plants under well-watered conditions and with freely orienting leaves were taller, and had a larger number of ramifications. The greater development favored the setting of a higher number of inflorescences per plant in these treatments. This behavior resulted in a high number of flowers, green and mature legumes per plant, thus resulting in high seed production which was the most evident response to water availability. Although individual seed weight was higher in the water stress treatment, total seed production was higher for well-watered plants, with no statistically significant effect of leaf movements.

Seed anatomy and germination of Phoenix roebelenii O'Brien (Arecaceae)

The objective of this study was to investigate the morphology, anatomy and germination behaviour of Phoenix roebelenii seeds. Biometric data were obtained by measuring 100 seeds extracted from recently harvested fruits and air-dried for one day. Four replications of 50 seeds each were previously treated with Vitavax-Thiran and then put to germinate in Sphagnum sp. in plastic trays at room temperature. Morphological details of the seeds were documented with the help of a scanning electronic microscope and then drawings were made with the help of a clear camera coupled to a stereomicroscope. Permanent lamina containing embryo sections were prepared to study its anatomy. The mean dimensions of the seeds were: length of 10.32mm, width of 5.21mm and thickness of 3.91mm. The weight of one thousand seeds was of 151.1g and the mean number of units.kg-1 was 6,600. Germination started between 27 and 58 days after sowing. The seeds are of the albuminous type, the endosperm is hard and the embryo (which is not clearly differentiated) occupies a lateral and peripheral position. During seed germination, seedling protrusion begins with the opening of an operculum, through which the cotyledon petiole is emitted with the embryonic axis at its tip. The portion of the cotyledon petiole that remains inside the seeds acts as a haustorium for the absorption of nutrients from the endosperm. The plumule emerges through a rift in the posterior part of the cotyledon. Secondary roots are observed to grow from the anterior part of the primary root.

Germination, initial growth and cotyledon protein content of bean cultivars under salinity stress

Excess salts in the root zone inhibit water uptake by plants, affect nutrient uptake and may result in toxicities due to individual salts in the soil solution. Excess exchangeable sodium in the soil may destroy the soil structure to a point where water penetration and root aeration become impossible. Sodium is also toxic to many plants. Beans (Phaseolus vulgaris L.) are consumed as protein source in northeastern Brazil, although little is known about common bean cultivar tolerance to salinity. The germination of bean cultivars under salt stress was studied. The cultivars 'Carioca' and 'Mulatinho' were submitted to germination test in a germinator at 25ºC, at the Seed Analysis Laboratory of the Brazilian Agricultural Research Corporation unit in the Semi- Arid region (Embrapa Semi Árido), Petrolina, Pernambuco State. These seeds were germinated on "germitest" papers imbibed in distilled water or in 10, 50, 100 e 200 mol.m-3sodium chloride (NaCl) solutions. At the first and second counts of the germination test, normal seedlings were counted, measured, weighed and dried, supplying data for vigor, total germination, fresh matter weight and dry matter weight and seedlings length. Total protein was quantified in cotyledons at 3, 6 and 9 days after sowing. The results indicated that the NaCl content influenced seed germination and concentrations above 50 mol.m-3 decreased germination and seedling growth.

Drying and germination of cupuassu (Theobroma grandiflorum (willd. Ex Spreng.) K. Schum.) seeds

The objective of this study was to verify the effect of drying on germination of cupuassu (Theobroma grandiflorum (Willd. ex Spreng) K. Schum.) seeds. Desiccation was in forced air oven, with temperature ranging from 23 to 33ºC. Sowing was carried out at 0.5cm of depth in plastic trays in sand and sawdust mixture (1:1), previously sterilized in hot water (100ºC), during 2h. Seeds were left to germinate in a laboratory with no temperature and relative humidity control, under natural light. It was quantified the seed moisture content, in four replications of 10 seeds; the germination percentage, performed during 30 days, with daily counts of the number of germinated seeds; the germination speed index; and number of days to the germination onset. The experimental design was completely randomized with four replications of 25 seeds. The reduction of moisture content from 58.6 to 37.8% did not affect seed germination and germination speed index; however, they were affected when moisture content was reduced to values below 30.7%. It was observed that only when moisture content was 16.1% seeds demanded more days to begin germination. Cupuassu seeds are classified as recalcitrant and they can be desiccated up to 37.8% with no reduction on germination.

Lolium multiflorum seeds in the soil: I. Soil seed bank dynamics in a no til system

The objective of the study was to characterize annual ryegrass seed population dynamics, managed for natural re-sowing, in no til systems in rotation with soybean, in different chronosequences An area was cultivated for two years with soybean, left as fallow land for the next two years and then cultivated again with soybean for the next two years. The four chronosequences represented different management periods, two with soybean (6 and 8 years old) and the other two resting (3 and 9 years old). Soil samples were taken every month during one year and divided into two depths (0-5 and 5-10 cm). Vegetation dynamics were also evaluated (number of plants, inflorescences and seedlings). Soil seed bank (SSB) dynamics showed structural patterns in time, with a "storage period" in summer, an "exhausting period" during autumn and a "transition period" in winter and spring. Pasture establishment by natural re-sowing was totally dependent on the annual recruitment of seeds from the soil. The influence of the management practices on the SSB was more important than the number of years that these practices had been implemented. Places where soybean was sown showed the largest SSBs. Most of the seeds overcame dormancy and germinated at the end of the summer and beginning of the autumn, showing a typically transitory SSB, but with a small proportion of persistent seeds

Lolium Multiflorum seeds in the soil: II. Longevity under natural conditions

The interest was the seed longevity dynamics of annual ryegrass in natural conditions as an important tool to explain its dynamics in no tillage systems used in the South of Brazil. The species is commonly managed for natural re-sowing and, in this way, allows cattle grazing with reduced costs during the winter time. In February of 2003, twenty bags of nylon screen containing sterilised soil with 100 seeds in each were randomly buried in the field, 5 cm deep. Around every three or four months, four sacks were exhumed. Seeds were counted and tested using germination and tetrazolium tests.The seeds showed high primary dormancy, which was overcome very fast. After 108, 226, 326, 565 and 732 days of burial there were no significant differences as the secondary dormancy of the seeds that did not germinate in autumn was not high. The last exhumation period was significantly different from the others due to the strong decay on seed viability. As few seeds remained viable after 732 days, the soil seed bank was classified as transient, being evident that in annual pastures the transitory seed banks have a main role in the regeneration of the species.