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Population developmental stage determines the recovery potential of Daphnia magna populations after fenvalerate application.

This study investigated the responses of Daphnia magna populations to pulsed exposures of the pyrethroid insecticide Fenvalerate applied during an early and a late stage of population development, and analyzed the dynamics of the subsequent recovery. A novel digital observation technique was used to describe the size and numbers of animals. High Fenvalerate concentrations caused high mortality rates during exponential population growth as well as during the food-limited stationary phase. However, recovery of populations took considerably longer in the stationary phase than in populations growing exponentially. The poor nutritional and reproductive state of food-deprived adults was indicated as the main cause of the slow recovery of populations. It is argued that populations operating at the carrying capacity of their environment are vulnerable to toxicant-induced disturbances to an extent not predictable from observations on exponentially growing populations such as are commonly used in ecotoxicology.

Animals↗

Identification of volatile compounds in cantaloupe at various developmental stages using solid phase microextraction.

Using an automated rapid headspace solid phase microextraction (SPME) method for volatile extraction in cantaloupes, 86 compounds already reported for muskmelons were recovered and an additional 53 compounds not previously reported were identified or tentatively identified. The SPME method extracted a copious number of volatiles that can be analyzed to clearly differentiate between variety, growth stage, and stage of harvest ripeness. Most of the newly reported compounds in cantaloupe were esters and aldehydes that have already been demonstrated as flavor-related compounds in other products. All esters believed to have flavor impact increased progressively after pollination, and this trend continued with increasing harvest maturity. However, compound recovery often decreased when fruits were harvested over-ripe. Most aldehydes increased during early growth stages and then tapered off with increasing harvest maturity. The SPME method suitably recovered most compounds reported to impart characteristic flavor/aroma in muskmelons. SPME offers experimental flexibility and the ability to discover more compounds and address flavor quality changes in fresh-cut cantaloupe.

Aldehydes↗

Lignin deposition and associated changes in anatomy, enzyme activity, gene expression, and ruminal degradability in stems of tall fescue at different developmental stages.

Stem tissues of tall fescue (Festuca arundinacea Schreb.) were sampled at three elongation stages and three reproductive stages. Anatomical analysis showed the deposition of guaiacyl (G) and syringyl (S) lignin during plant development and the formation of a lignified sclerenchyma ring. A dramatic increase in Klason lignin content was found from elongation stage to reproductive stage. Lignin composition analyzed by gas chromatography-mass spectrometry revealed that S lignin content and S/G ratio increased with stem development, but contents of p-hydroxyphenyl (H) and G lignins decreased during the same period. S lignin content and S/G ratio also increased from the younger upper internode down to the older basal internode of the stem, but G and H lignin decreased in parallel. Relative O-methyltransferase activities increased during stem development and in parallel with the lignification process of stem. The pattern of enzyme activity during development varied with the choice of substrate, with highest activities seen when substrates were caffeoylaldehyde and 5-hydroxyferulic acid, and lowest activities were seen when caffeic acid and 5-hydroxyconiferyl alcohol were used as substrates. The expression of caffeic acid O-methyltransferase and cinnamyl alcohol dehydrogenase genes increased during the stem elongation stage and remained at high levels during the reproductive stages. The changes at anatomical, metabolic, and molecular levels during plant development were closely associated with lignification and degradability. This study provides an integrated picture of the molecular and chemical events that accompany changes in lignin deposition and ruminal degradability.

Alcohol Oxidoreductases↗

Antioxidant activity in fruits and leaves of blackberry, raspberry, and strawberry varies with cultivar and developmental stage.

Fruits and leaves from different cultivars of thornless blackberry (Rubus sp.), red raspberry (Rubus idaeus L.), black raspberry (Rubus occidentalis L.), and strawberry (Fragaria x ananassa D.) plants were analyzed for total antioxidant capacity (oxygen radical absorbance capacity, ORAC) and total phenolic content. In addition, fruits were analyzed for total anthocyanin content. Blackberries and strawberries had the highest ORAC values during the green stages, whereas red raspberries had the highest ORAC activity at the ripe stage. Total anthocyanin content increased with maturity for all three species of fruits. Compared with fruits, leaves were found to have higher ORAC values. In fruits, ORAC values ranged from 7.8 to 33.7 micromol of Trolox equivalents (TE)/g of fresh berries (35. 0-162.1 micromol of TE/g of dry matter), whereas in leaves, ORAC values ranged from 69.7 to 182.2 micromol of TE/g of fresh leaves (205.0-728.8 micromol of TE/g of dry matter). As the leaves become older, the ORAC values and total phenolic contents decreased. The results showed a linear correlation between total phenolic content and ORAC activity for fruits and leaves. For ripe berries, a linear relationship existed between ORAC values and anthocyanin content. Of the ripe fruits tested, on the basis of wet weight of fruit, cv. Jewel black raspberry and blackberries may be the richest source for antioxidants. On the basis of the dry weight of fruit, strawberries had the highest ORAC activity followed by black raspberries (cv. Jewel), blackberries, and red raspberries.

Anthocyanins↗

Proteomic analysis of the silkworm (Bombyx mori L.) hemolymph during developmental stage.

We utilized the proteomic approach to investigate the proteome of the fifth instar hemolymph during growth and development, and to improve the understanding of this important bioprocess and gene expression situation. A total of 25 microL of hemolymph was used for 2D analysis, and the separated proteins were visualized by silver staining and analyzed using the ImageMaster 2D software. The report showed as many as 241 of protein spots were expressed in the beginning of the fifth instar. Among them, most were concentrated in pI 3.5-6.5, which reached 76% of the total protein spots. As for the protein molecular sizes, 182 protein spots concentrated between 35 and 90 kDa, which comes to 75% of the total spots. When the larvae grow to the seventh day (total fifth instar duration was 9 days), 298 protein spots were visualized through 2D electrophoresis. Fifty-seven spots were newly expressed compared to the image of the first day in fifth instar. The results implied that these proteins are related to biosynthesis of silk protein and metamorphosis preparation from larva to pupa. In total, 19 protein spots including 6 special spots expressed in seventh day were analyzed through MALDI-TOF-MS. The relations between proteins and growth and development of silkworm were discussed.

Amino Acid Sequence↗

Differential effects of a six-day immobilization on newborn rat soleus muscles at two developmental stages.

Our objective was to determine the effects of a six-day immobilization on the musculoskeletal system of the rat during postnatal development at two key periods when the states of innervation are known to be different. This work was undertaken on the soleus muscle since it is well known that postural slow muscles show marked changes after a period of disuse. Thus, the soleus muscle was immobilized in a shortened position either when the innervation was polyneuronal or monosynaptic, respectively from 6 to 12 and from 17 to 23 days. The muscle modifications were followed by ATPase staining and myosin heavy chain (MyHC) isoform identification using monoclonal antibodies and SDS-PAGE. The functional properties of skinned fibre bundles were established by calcium/strontium (Ca/Sr) activation characteristics. In control muscles the maturation was characterized by a progressive increase of adult MyHCs (I and IIA) concomitant with a decrease in both the MyHC neo and the Ca affinity. Between 6 to 12 days, immobilization of the limb induced an increase in histochemical type IIC fibres. Using antibodies we identified new fibre types, classified as a function of their MyHC isoform co-expression. We observed an increase in expression of both MyHC neo and Ca affinity. From 17 to 23 days, the immobilization induced an increase in Ca affinity and marked changes in the MyHC isoform composition: disappearance of MyHC neo and expression of the fast MyHC IIB isoform, which in normal conditions is never expressed in the soleus muscle. We conclude that an immobilization imposed during polyneuronal innervation delays the postnatal maturation of the soleus muscle, whereas when the immobilization is performed under monosynaptic innervation the muscle evolves towards a fast phenotype using a default pathway for MyHC expression.

Adenosine Triphosphatases↗

Immunohistochemical study of glutathione reductase in rat ocular tissues at different developmental stages.

Glutathione, which is found in high levels in eye tissues, is involved in multiple functions, including serving as an antioxidant and as an electron donor for peroxidases. Although the activities of enzymes related to glutathione metabolism have been reported in the eye, the issue of which cells produce these proteins, where they are produced and at what levels is an important one. Glutathione reductase, an enzyme which recycles oxidized glutathione by transferring electrons from NADPH, was localized immunohistochemically in adult rat eye in this study. The reductase was distributed in the corneal and conjunctival epithelia, corneal keratocytes and endothelium, iridial and ciliary epithelia, neural retina, and retinal pigment epithelium. In addition, it was highly expressed in ganglion cells, which are responsible for transmitting photophysiological signals from the retina to the higher visual centres. To clarify the correlation of glutathione reductase expression and oxidative stress, the enzymatic activity and the level of protein expression at the pre- and postnatal stages was examined. Expression of the enzyme was detected first in the ganglion cell layer of a late prenatal stage, and appeared in the inner plexyform layer after birth. Along with an increasing differentiation between the inner nuclear and outer nuclear layers, glutathione reductase expression became detectable in the outer plexyform layer. Pigment epithelial cells were positively stained only after birth. Expression was also detected in the lens epithelium from the prenatal to early postnatal stages although its level was low in the adult lens. Collectively, these data, except for lens epithelia, suggest the pivotal role of glutathione reductase in recycling oxidized glutathione for the protection of the tissues against oxidative stress, which is caused by eye opening accompanied by the initiation of various ocular processes, such as accession of light and transduction of the photochemical signal.

Animals↗

Characterization of the developmental stages of cypress zygotic embryos by two-dimensional electrophoresis and by cytochemistry.

Biochemical changes that characterize megagametophyte and zygotic embryo development in the conifer Cupressus sempervirens L. (Cupressaceae) were studied by complementary methods of cytochemistry and two-dimensional electrophoresis (2-DE). These analyses revealed that early in their development megagametophytes and embryos were characterized by the predominant elaboration of starch in association with a low protein content. As their development proceeded, starch content gradually decreased while protein body synthesis progressively intensified, both in the megagametophyte and the embryo. In parallel, an increase in protein level as well as an accumulation of specific polypeptides could be observed in the two tissues. During maturation, protein bodies accumulated to high levels both in megagametophyte and embryo cells, whereas starch could no longer be detected. Protein levels were high in mature seeds and reached 12% and 8% of the megagametophyte and embryo DW, respectively. Some sets of polypeptides accumulated more specifically at this time in both megagametophyte and embryo. Some of these began to first accumulate in the megagametophyte during embryo development before their concentration rose in the embryo at cotyledonary stage. Others displayed a more specific-embryo accumulation pattern.

Journal Article↗

Morphological spelling strategies: developmental stages and processes.

The spelling of many words in English and in other orthographies involves patterns determined by morphology (e.g., ed in past regular verbs). The authors report a longitudinal study that shows that when children first adopt such spelling patterns, they do so with little regard for their morphological basis. They generalize the patterns to grammatically inappropriate words (e.g., sofed for soft). Later these generalizations are confined to the right grammatical category (e.g., keped for kept) and finally to the right group of words (regular verbs). The authors conclude that children first see these spelling patterns merely as exceptions to the phonetic system and later grasp their grammatical significance. The study included two new measures of grammatical awareness, both involving analogies, that predicted success with spelling inflectional morphemes in later sessions.

Awareness↗

Characterization of translation systems in vitro from three developmental stages of Strongylocentrotus purpuratus.

We have developed and characterized cell-free systems active in translation from unfertilized eggs, 30-min zygotes and hatched blastulae of the sea urchin Strongylocentrotus purpuratus. The ion concentrations selected for preparation of the lysates were 150 mM-K+, 40 mM-Na+, 40 mM-Cl-, 5 x 10(-7) M free Ca2+ and 1 mM free Mg2+. It was necessary to include the ribonuclease inhibitor RNas in the preparations to obtain full activity consistently. The pH optimum was 7.2 and was extremely sharp for the three S. purpuratus lysates. The temperature optima of the three lysates were remarkably similar to those of the intact unfertilized egg and embryos. Lysates from unfertilized egg and 30-min zygotes showed a temperature optimum at 15 degrees C. The hatched blastula lysate showed a broader temperature optimum with a shift to about 20 degrees C. The optimized lysates incorporated radiolabelled amino acids into polypeptides for up to 90 min. The polypeptides synthesized ranged in Mr from 200,000 to 20,000, suggesting that the mRNA in the lysates was intact and capable of directing the synthesis of complete polypeptides. Furthermore, the three lysates were capable of initiation, as demonstrated by inhibition of initiation using the inhibitors edeine and 7-methylguanosine 5'-triphosphate (m7GTP). At 15 degrees C, the transit times for the three lysates were: unfertilized egg, 40 min; 30-min zygotes and hatched blastula lysates, 20 min. These transit times are similar to those of intact eggs and embryos, and significantly, reflect the two-fold increase in elongation rate seen following fertilization in intact embryos. Thus, these lysates display many features and characteristic responses typical of intact eggs and embryos, indicating that the lysates should be useful tools for the analysis of translation control in early embryogenesis.

Animals↗

Phosphorylation differences among proteins of bloodstream developmental stages of Trypanosoma brucei brucei.

Early in an infection the bloodstream forms of the African trypanosome Trypanosoma brucei brucei are long, slender and rapidly dividing. Later, non-dividing, short, stumpy forms may be found. In this report we described biochemical differences between the two parasite populations in the phosphorylation of their proteins in vitro. Compared with the slender populations, the non-dividing stumpy forms of the parasites exhibit decreased phosphorylation of an 80 kDa protein and enhanced phosphorylation of 37 kDa and 42 kDa proteins (pp37 and pp42). These changes occurred regardless of whether the stumpy trypanosomes were generated naturally during the course of the infection or induced by difluoromethylornithine treatment. The phosphorylation of pp37 and pp42 occurs on serine and threonine residues and is totally dependent upon the presence of Mn2+ or Mg2+. However, excess Mn2+ or Mg2+ inhibits phosphorylation. Maximal phosphorylation of pp42 occurs with 1 mm-Mn2+ or 10 mm-Mg2+, whereas that of pp37 occurs with 50 mM-Mn2+ or greater than 100 mm-Mg2+. The phosphorylation of pp37 is greatly enhanced by KCl, whereas that of pp42 is only slightly increased by this salt. Ca2+, calmodulin, phospholipids and cyclic AMP have no discernible effect upon the phosphorylation of pp42 or pp37 in vitro, whereas heparin, suramin, polylysine, polyarginine and polyamines all inhibit phosphorylation. Thus the enzymes that phosphorylate pp42 and pp37 have properties similar to, but distinct from, those of mammalian casein kinase II. Since the casein-kinase-like activity is higher in the slender than in the stumpy forms, the enhanced phosphorylation of pp42 and pp37 in the non-dividing parasites is probably a result of the enhanced synthesis of these acidic proteins.

Animals↗

Tissue- and developmental-stage-specific methylation in the two kidney promoters of the rat gamma-glutamyl transpeptidase gene.

We have investigated, using DNA methylation patterning, the site-specific methylation of promoters I and II of the rat gamma-glutamyl transpeptidase gene. This analysis was done in fetal, newborn and adult rat kidney, in which promoters I and II are progressively active during development, as well as in rat liver, which never expresses mRNAs from these two promoters. During kidney development, a progressive demethylation occurs in the promoter I and II region, specially at the level of the most proximal MspI site of promoter II. A progressive reorganization of the methylated sites within the 5' end of the gene also occurs during liver development.

Animals↗