Genetical and biochemical comparisons of alcohol dehydrogenase isozymes from Anastrepha fraterculus and A. obliqua (Diptera: Tephritidae): Evidence for gene duplication

The electrophoretic patterns of the enzyme alcohol dehydrogenase (ADH) from Anastrepha fraterculus and A. obliqua were studied. Two loci were found to code for the enzyme in A. fraterculus, and three in A. obliqua. In both species, all isozymes were active in third-instar larvae. A cationic isozyme (Adh-1) was active mainly in the visceral fat body of both species. In A. fraterculus, the locus had an anionic polymorphic isozyme (Adh-3) that was detected in the parietal fat body. In addition to these two loci, a third locus for an anionic isozyme (Adh-2), which was active in the digestive tube of larvae, was present in A. obliqua and probably resulted from gene duplication. For both species, multiple forms of the isozymes are formed by binding of an NAD-carbonyl compound, as in Drosophila melanogaster. Both larvae and early pupae of A. obliqua had almost twice the specific ADH activity as A. fraterculus. The ethanol content of the host fruit infested with A. obliqua (red mombim) was also higher than that of the host fruit infested with A. fraterculus (guava).This research was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnologico (CNPq-PIG 40.2486/82).     

Effect of Temperature on Development and Population Growth of Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae) on Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

The parasitic mite Acarophenax lacunatus kills the eggs upon which it feeds and seems to have potential as a biological control agent of stored grain pests. The lack of biological studies on this mite species led to the present study carried out in laboratory conditions at eight different temperatures (ranging from 20 to 41°C) and 60% relative humidity using Rhyzopertha dominica as host. The higher the temperature, the faster: (1) the attachment of female mites to the host egg (varying from 1 to 5 h); (2) the increase in body size of physogastric females (about twice faster at 40°C than at 20°C); and (3) the generation time (ranging from 40 to 220 h). In addition, the higher the temperature, the shorter the maximum female longevity (ranging from about 75 to 300 h). The two estimated temperature thresholds for development of A. lacunatus on R. dominica were 18 and 40°C. The average number of female and male offspring per gravid mite were 12.8 and 1.0, respectively, with sex ratios (females/total) ranging from 0.91 to 0.94 (maximum at 30°C). The net reproductive rate and intrinsic rate of increase also presented maximum values at 30°C (12.1 and 0.04, respectively).     

Growth kinetics of hemopoietic fetal stem cells

Fetal spleen stem cells have growth characteristics similar to those of normal adult spleen stem cells. On the contrary there is an early fetal liver stem cell population which possesses a lag time longer than that of adult stem cells. The duration of the lag time is controlled by a built-in biological timer which seems to regulate some proliferative functions of the primitive liver stem cell.     

Mathematical modelling and kinetic study for CD production catalysed by Toruzyme® and CGTase from Bacillus firmus strain 37

A new mathematical model was developed for the kinetics of α-, β- and γ-cyclodextrin production, expanding an existing model that only included the production of β- and γ-cyclodextrins, because a detailed kinetic modelling of the reactions involved allows the manipulation of the process yields. The kinetic behaviour of the commercial enzyme Toruzyme^® was studied with maltodextrin as substrate at different concentrations and for CGTase from Bacillus firmus strain 37 at a concentration of 100 g L⁻¹. The mathematical model showed a proper fit to the experimental data, within the 24-h period studied, confirming that the considered hypotheses represent the kinetic behaviour of the enzymes in the reaction medium. The kinetic parameters generated by the model allowed reproducing previous observed qualitative tendencies as it can be seen that changing experimental conditions in the reaction process such as enzyme and substrate concentrations results in large changes in the enzyme kinetics and using high substrate concentrations does not guarantee the highest conversion rates due to enzyme inhibition and reverse reactions. In addition, this new mathematical model complements previous qualitative observations enabling the manipulation of the direct and reverse reactions catalysed by the enzyme by adjusting the reaction conditions, to target quantitative results of increased productivity and better efficiency in the production of a desired cyclodextrin.

     

Group I introns and associated homing endonuclease genes reveals a clinal structure for <Emphasis Type="Italic">Porphyra spiralis</Emphasis> var. <Emphasis Type="Italic">amplifolia</Emphasis> (Bangiales,Rhodophyta) along the Eastern coast of South America

Background  

Group I introns are found in the nuclear small subunit ribosomal RNA gene (SSU rDNA) of some species of the genus Porphyra (Bangiales, Rhodophyta). Size polymorphisms in group I introns has been interpreted as the result of the degeneration of homing endonuclease genes (HEG) inserted in peripheral loops of intron paired elements. In this study, intron size polymorphisms were characterized for different Porphyra spiralis var. amplifolia (PSA) populations on the Southern Brazilian coast, and were used to infer genetic relationships and genetic structure of these PSA populations, in addition to cox 2-3 and rbc L-S regions. Introns of different sizes were tested qualitatively for in vitro self-splicing.     

Hemopoietic stem cell distribution in tissues of fetal and newborn mice

The variations in the distribution of hemopoietic stem cells in fetal tissues are concomitant with changes in the electrophoretic pattern of hemoglobin. The data presented here are consistent with the hypothesis that the embryonic stem cells can be transformed into adult stem cells in the liver of 15 to 16 days old fetuses.     

Ultrafiltration system for cyclodextrin production in repetitive batches by CGTase from <Emphasis Type="Italic">Bacillus firmus</Emphasis> strain 37

This study aimed to improve the yield of cyclodextrins (CDs) production in repetitive batches. An innovative ultrafiltration system was used to remove the inhibitory products that accumulated in the medium and to recover the enzyme. The assays were performed with the CGTase from Bacillus firmus strain 37 in purified, semi-purified, and crude extract forms. Maltodextrin (10 % w/v) and corn starch (5 % w/v) were used as substrates. After eight repetitive 24-h batches, the yield of β-CD obtained with the purified enzyme and the corn starch substrate was 0.54 mmol/L/h, which was 36 % greater than that observed with the 10 % maltodextrin substrate. The crude CGTase extract with the corn starch substrate showed a productivity of 0.38 mmol/L/h, which was 29 % lower than using the purified enzyme and the corn starch substrate but 7 % higher than using the purified enzyme and the maltodextrin substrate. The crude extract, assayed with the corn starch substrate in the presence of 10 % ethanol reached 0.43 mmol/L/h productivity, which was 12 % higher compared to the assay without ethanol. The semi-purified enzyme was assayed with the corn starch substrate in the presence of 10 % ethanol for eight batches lasting 12 h and an excellent selectivity for the β-CD was obtained, reaching a mean percentage of 96.0 %. Therefore, this ultrafiltration system enabled several batches of CD production, with efficient removal of products inhibitory to the CGTase and recovery of the enzyme. The possibility of industrial application of this system is promising.     

Mating time of the West Indian fruit fly Anastrepha obliqua (Macquart) (Diptera: Tephritidae) under laboratory conditions

Allochronic reproductive isolation seems to be an important factor in speciation processes in Tephritidae since specific mating times are a widespread feature of its species. The timing of matings of the West Indian fruit fly, Anastrepha obliqua (Macquart) was investigated through group-focal observations, during ten days, under laboratory conditions. The number of observed matings and males exhibiting calling behavior varied significantly according to time of day. Sexual activities seemed to be concentrated in the afternoon period, with the male calling behavior reaching a peak between 3:30p.m. and 4:30p.m., and mating occurred most frequently from 2:00 p.m. to 4:30 p.m.