Independent differentiation of mammotropes and somatotropes in the chicken embryonic pituitary gland

It has been reported that mammotropes in a rodent pituitary gland are derived from somatotropes via somatomammotropes (SMTs), cells that produce both growth hormone (GH) and prolactin (Prl). However, no studies have been done on the transdifferentiation of somatotropes in the chicken pituitary gland. In this study, in order to determine the origin of mammotropes, we studied detail property of appearance of chicken somatotropes, mammotropes and pit-1 cells and then evaluated the existence of SMTs in the chicken embryonic pituitary gland. Immunohistochemical analysis revealed that GH-immunopositive (GH-ip) cells appeared on embryonic day (E) 14 and were mainly distributed in the caudal lobe, while Prl-immunopositive (Prl-ip) cells appeared in the cephalic lobe of the pituitary gland on E16. In situ hybridization (ISH) and RT-PCR analysis showed that expression of GH and Prl mRNA starts at E12 in the caudal lobe and at E14 in the cephalic lobe respectively. Pit-1 mRNA was first detected on E5 by RT-PCR, and pit-1 mRNA-expressing cells were found in the cephalic lobe on E8. Then with the ontogeny of the chicken, these cells spread into both lobes. Using a double staining method with ISH and immunohistochemistry, we could not detect the existence of SMTs in the chicken embryonic pituitary gland even in the marginal region of either lobe. These results suggest that chicken somatotropes and mammotropes independently appear in different lobes of pituitary gland and that transdifferentiation from somatotropes to mammotropes is not the central route for differentiation of mammotropes in the embryonic chicken pituitary gland.     

Size-selective dispersal of Daphnia resting eggs by backswimmers (Notonecta maculata)

Freshwater zooplankton is increasingly used to study effects of dispersal on community and metacommunity structure. Yet, it remains unclear how zooplankton disperses. Clearly, birds and wind play a significant role as zooplankton dispersal agents, but they may not always be the main vectors. This experimental study shows that a cosmopolitan aquatic insect, Notonecta, can be an important vector of cladoceran resting eggs (ephippia). Dispersing Notonecta frequently transported ephippia during flight, with a bias towards smaller ephippia in two species. A similar trend was present at the species level: Daphnia species with smaller ephippia were more often dispersed, suggesting that Notonecta could generate specific colonist communities. In addition, buoyancy appeared a critical trait, as non-floating ephippia of Daphnia magna were never dispersed. Our data suggest that Notonecta could be important dispersers of Daphnia, and that knowledge of dispersal dynamics of Notonecta may be used to predict Daphnia dispersal, colonization and resilience to disturbance.     

Expanded bed adsorption as a unique unit operation for the isolation of bacteriocins from fermentation media

Expanded bed adsorption using a strong cation exchanger allowed the direct isolation of amylovorin L471, a bacteriocin from Lactobacillus amylovorusDCE 471, from the fermentation medium. The pH of the loading and elution buffer were optimised in a packed bed with cell-free culture supernatant. Bound bacteriocin was eluted with 1.0 M NaCl. The highest recovery (30%) was obtained at the lowest pH (3.6). At higher pH values the recovery was lower, namely 12%, 15% and 7% at pH 4.5, 6.5 and 8.0, respectively. In expanded bed mode, direct isolation of the bacteriocin from the fermentation medium at pH 3.6 (loading and elution) initially resulted in a recovery of 12%. After optimisation of the pH (loading and elution at pH 3.6 and 6.5, respectively), the recovery for amylovorin L471 increased up to 30% and higher. Recovery of enterocin A from Enterococcus faeciumCTC 492 fermentation medium averaged 15% (loading and elution at pH 3.6 and 6.0, respectively). With pediocin, produced by Pediococcus acidilactici ATCC 8042, 26% recovery was obtained at a pH of 6.5 during loading and elution. Low recoveries can be ascribed to non-optimal operation conditions (pH of loading and elution buffer), inactivation of the bacteriocin on a cationic resin, and the formation of more insoluble and less active, strongly hydrophobic bacteriocin aggregates upon further purification.     

Breathing pattern in highly competitive cyclists during incremental exercise

The purpose of our investigation was to analyse the breathing patterns of professional cyclists during incremental exercise from submaximal to maximal intensities. A group of 11 elite amateur male road cyclists [E, mean age 23 (SD 2) years, peak oxygen uptake (VO2peak) 73.8 (SD 5.0) ml kg(-1) min(-1)] and 14 professional male road cyclists [P, mean age 26 (SD 2) years, (VO2peak) 73.2 (SD 6.6) ml kg(-1) min(-1)] participated in this study. Each of the subjects performed an exercise test on a cycle ergometer following a ramp protocol (exercise intensity increases of 25 W x min(-1)) until the subject was exhausted. For each subject, the following parameters were recorded during the tests: oxygen consumption (VO2), carbon dioxide output (VCO2), pulmonary ventilation (VE), tidal volume (VT), breathing frequency (fb), ventilatory equivalents for oxygen (VE x VO2(-1)) and carbon dioxide (VE x VCO2(-1)), end-tidal partial pressure of oxygen and partial pressure of carbon dioxide, inspiratory (tI) and expiratory (tE) times, inspiratory duty cycle (tI/tTOT, where tTOT is the time for one respiratory cycle), and mean inspiratory flow rate (VT/tI). Mean values of VE were significantly higher in E at 300, 350 and 400 W (P < 0.05, P < 0.05 and P < 0.01, respectively); fb was also higher in E in most moderate-to-maximal intensities. On the other hand, VT showed a different pattern in both groups at near-to maximal intensities, since no plateau was observed in P. The response of tI and tE was also different. Finally, VT/tI and tI/tTOT showed a similar response in both P and E. It was concluded that the breathing pattern of the two groups differed mainly in two aspects: in the professional cyclists, VE increased at any exercise intensity as a result of increases in both VT and fb, with no evidence of tachypnoeic shift, and tE was prolonged in this group at high exercise intensities. In contrast, neither the central drive nor the timing component of respiration seem to have been significantly altered by the training demands of professional cycling.     

Reducing mutational bias in random protein libraries

The success of protein optimization through directed molecular evolution depends to a large extent on the size and quality of the displayed library. Current low-fidelity DNA polymerases that are commonly used during random mutagenesis and recombination in vitro display strong mutational preferences, favoring the substitution of certain nucleotides over others. The result is a biased and reduced functional diversity in the library under selection. In an effort to reduce mutational bias, we combined two different low-fidelity DNA polymerases, Taq and Mutazyme, which have opposite mutational spectra. As a first step, random mutants of the Bacillus thuringiensis cry9Ca1 gene were generated by separate error-prone polymerase chain reactions (PCRs) with each of the two polymerases. Subsequent shuffling by staggered extension process (StEP) of the PCR products resulted in intermediate numbers of AT and GC substitutions, compared to the Taq or Mutazyme error-prone PCR libraries. This strategy should allow generating unbiased libraries or libraries with a specific degree of mutational bias by applying optimal mutagenesis frequencies during error-prone PCR and controlling the concentration of template in the shuffling reaction while taking into account the GC content of the target gene.     

Morphometric analyses of Staurosira inflata comb. nov. (Bacillariophyceae) and the morphologically related Staurosira tabellaria from north-western Russia

The morphology and ontogenetic allometric trends of a rare diatom Fragilaria heidenii Østrup and the morphologically related Staurosira tabellaria (W. Smith) Leuduger-Fortmorel were compared using conventional and semilandmark-based geometric morphometric analyses. Fragilaria heidenii was studied in detail by light and electron microscopy using type material and recent samples from Lake Ladoga and Lake Ilmen (north-western Russia). The taxon is transferred into the genus Staurosira Ehrenberg as Staurosira inflata comb. nov. on the basis of its valve morphology. This taxon is characterized by the absence of rimoportulae, lack of perforated copulae, spines located on the interstriae, internal vola occlusion in the areolae and the features of the areolae and apical pore fields. Conventional morphometric analysis showed considerable overlapping of S. inflata and S. tabellaria in their frustule characteristics such as length, width, length-to-width ratio and striae density. Moreover, at later stages of the vegetative life cycle, S. inflata has a tendency to resemble S. tabellaria by its valve outline that makes it difficult to separate these two taxa. The geometric morphometric analysis revealed two shape groups corresponding to S. inflata and S. tabellaria that were separated by a clear gap. Semilandmarks representing shape of the middle part of the valve were primarily responsible for discrimination between species. Apart from differences in valve shapes, S. inflata and S. tabellaria also differed significantly in their ontogenetic allometric trajectories. Overall, our results demonstrate that the semilandmark-based geometric morphometrics is sensitive enough to distinguish species by their outlines, when traditional morphometric parameters are not able to discriminate them with confidence.     

Balancing competition for resources with multiple pest regulation in diversified agroecosystems: a process‐based approach to reconcile diversification and productivity

Agroecosystem plant diversification can enhance pest biological regulation and is a promising alternative to pesticide application. However, the costs of competition for resources between plants may exceed the benefits gained by pest regulation. To disentangle the interactions between pest regulation and competition, we developed a generic process‐based approach that accounts for the effects of an associated plant and leaf and root pests on biomass production. We considered three crop–plant associations that differ in competition profiles, and we simulated biomass production under wide ranges of both pest regulation rates and resources’ availability. We analyzed outputs to quantify the pest regulation service level that would be required to attain monoculture yield and other production goals. Results showed that pest regulation requirements were highly dependent on the profile of resource interception of the associated plant and on resources’ availability. Pest regulation and the magnitude of competition between plants interacted in determining the balance between nitrogen and radiation uptake by the crop. Our findings suggest that productivity of diversified agroecosystems relative to monoculture should be optimized by assembling plants whose characteristics balance crops’ resource acquisition. The theoretical insights from our study draw generic rules for vegetation assemblage to optimize trade‐offs between pest regulation and production. Our findings and approach may have implications in understanding, theorizing and implementing agroecosystem diversification. By its generic and adaptable structure, our approach should be useful for studying the effects of diversification in many agroecosystems.     

Influence of diet on life table parameters of <Emphasis Type="Italic">Iphiseius degenerans</Emphasis> (Acari: Phytoseiidae)

Studies on the reproduction, longevity and life table parameters of Iphiseius degenerans (Berlese) were carried out under laboratory conditions of 25 ± 1 °C, 75 ± 5% RH and 16L:8D h. As food sources for the predatory mite, Ricinus communis L. pollen, all stages of the spider mite Tetranychus urticae Koch, Frankliniella occidentalis (Pergande) larvae, and Ephestia kuehniella Zeller eggs were selected. All diets were accepted as food by the adult mites. Female longevity ranged from 29.5 to 42.4 days, the highest value was recorded on a diet of Ephestia eggs. The highest percentage of females escaping the experimental arena was observed on the diet consisting of thrips larvae. The highest oviposition rate (1.9 eggs/female.day) was recorded when the predator was fed on spider mites on an artificial substrate. For other diets, oviposition rates ranged from 1.0 to 1.3 eggs/female.day. The intrinsic rate of natural increase (r _m) of I. degenerans varied between 0.015 and 0.142 females/female.day. The diet consisting of castor bean pollen resulted in the highest population growth whereas the diet on spider mites brushed off onto a bean leaf arena resulted in the slowest population growth. This can be explained by the inability of the predator to cope with the webbing of T. urticae, and the high escape rate of the progeny when reared on spider mites. The percentage of females in the offspring ranged from 40 to 73%.This revised version was published online in May 2005 with a corrected cover date.