Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production

It has been demonstrated that caleosin alone is sufficient to stabilize artificial oil bodies. A series of recombinant caleosins, mutated with 3, 5, 8, 11, 13, 15, and 17 extra Lys residues and over‐expressed in Escherichia coli, were used as carrier proteins to render biotin as a hapten on the surface of artificial oil bodies for antibody production. Biotinylation levels of the recombinant caleosins were step‐wisely elevated as the number of extra Lys residues increased, and the biotinylated Lys residues were identified by mass spectrometric analysis. Polyclonal antibodies against biotin were successfully generated in rats injected with artificial oil bodies constituted with each of the biotinylated caleosins. Moreover, those generated via the biotinylated caleosins with eight or more extra Lys residues no longer recognized caleosin. It appears that engineered Lys‐rich caleosins are suitable carrier proteins for the production of antibodies against small molecules. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011     

Using Echo Ultrasound from Schooling Fish to Detect and Classify Fish Types

Fish finders have already been widely available in the fishing market for a number of years.However,the sizes of these fishfinders are too big and their prices are expensive to suit for the research of robotic fish or mini-submarine.The goal of thisresearch is to propose a low-cost fish detector and classifier which suits for underwater robot or submarine as a proximity sensor.With some pre-condition in hardware and algorithms,the experimental results show that the proposed design has good per-formance,with a detection rate of 100 % and a classification rate of 94 %.Both the existing type of fish and the group behaviorcan be revealed by statistical interpretations such as hovering passion and sparse swimming mode.     

The effects of different PMSG doses on estrus behavior and pregnancy rate in Angora goats

Artificial insemination protocols depend on efficient behavioral estrus detection and insemination time in Angora goat. Therefore, we aim to determine the accuracy of an estrus scoring system in Angora goats with different PMSG doses during the breeding season. Does (n: 260) were randomly divided into three groups: group-1 (n: 93), group-2 (n: 85) and group-3 (n: 82). All animals received an intravaginal sponge on day 0 for 11 days, and on the day of sponge insertion 150 μg prostaglandin F2Α was administered. Pregnant mare’s serum gonadotropin was injected 300, 400 and 500 IU intramuscularly 24 h before sponge removal to groups 1, 2 and 3, respectively. Estrus signs were detected with a teaser buck, 24 h after sponge removal according to a visual scoring system. Artificial insemination was performed with 0.25 ml fresh diluted semen at 43 to 45 h after sponge removal. Differences were observed within PMSG groups in terms of standing, tail wagging, courtship behavior, vaginal discharge and vaginal hyperemia (P<0.001). Nevertheless, the most accurate indicators of estrus that result in pregnancy were tail wagging and courtship behavior followed by standing estrus (P<0.05). According to the results obtained, 300 IU PMSG dose is sufficient, both to inseminate at a fixed time (43 to 45 h after sponge removal) and to record the estrus behavior by teaser male 24 h after sponge removal. Higher PMSG doses (400 to 500 IU) altered the timing of ovulation; specifically, 500 IU dose shortened the duration of estrus behaviors. In conclusion, even though the different doses of PMSG displayed similar effects on estrus synchronization and pregnancy rates, we concluded that tail wagging, courtship behavior and standing heat are the most reliable estrus signs for artificial insemination in Angora goat.     

Validation of the first step of the heuristic refinement method for the derivation of solution structures of proteins from NMR data

A new method for the analysis of NMR data in terms of the solution structure of proteins has been developed. The method consists of two steps: first a systematic search of the conformational space to define the region allowed by the initial set of experimental constraints, and second, the narrowing of this region by the introduction of additional constraints and optional refinement procedures. The search of the conformational space is guided by heuristics to make it computationally feasible. The method is therefore called the heuristic refinement method and is coded in an expert system called PROTEAN. The paper describes the validation of the first step of the method using an artificial NMR data set generated from the known crystal structure of sperm whale carbon monoxymyoglobin. It is shown that the initial search procedure yields a low-resolution structure of the myoglobin molecule, accurately reproducing its main topological features, and that the precision of the structure depends on the quality of the initial data set.     

Life cycle changes during 12 successive generations of laboratory colonisation of the leafroller Cnephasia jactatana (Walker) (Lepidoptera: Tortricidae) on artificial diet

Abstract

Close monitoring of the lepidopteran leafroller Cnephasia jactatana under laboratory colonisation revealed few distinct effects of successive rearing on artificial diet on the life cycle. The second laboratory generation had a prolonged development time and altered sex synchronism in pupation and eclosion patterns. Some deleterious changes were observed in later generations, including decreases in fertility, egg hatch and sperm motility, failure of mating adults to separate, and pupal and adult malformations. These changes were not adaptive, but were due to incompatibility with the general purpose diet (GPD) used; they were absent under sub-colonisation on a sheepnut-bean based diet (SBD). Success in the laboratory colonisation of C. jactatana is attributed to a random mating protocol, choice of environmental conditions representing the wild habitat, and a rapid rate of population growth.     

Determination of adenosine triphosphate in human semen to estimate the fertilizing potential and to quantify sperm antibodies

The measurement of the ATP content of fresh semen is as accurate as the estimation of sperm motility by conventional methods in discriminating between semen of fertile versus subfertile men. The ATP content of frozen thawed donor semen is correlated with the probability of conception per cycle of insemination. Exact quantification of cytotoxic sperm antibodies in serum is possible with the adenosine-triphosphate-release-cytotoxicity test, since measurement is free of the bias of microscopic examination. The procedure has been simplified by testing only one serum dilution and calculating the ‘sperm toxicity index’.     

Artifical diets and rearing of the nun moth, Lymantria monacha

Artificial diets were reviewed and two tested. The highest level of survival to the adult stage (72%) was obtained on a modified diet of Odell & Rollinson (1966). Rearing required 61 days for males and 70 days for females. Wing deformation occurred in 16% of the adults. The sex ratio ( : ) was 0.80. Average pupal weight was 281 mg for males and 368 mg for females. Females averaged 102 eggs (range 80–125 eggs). Neonates from field-collected egg masses started hatching on 6 December; frost was not necessary for hatching. Hatching could be postponed until at least 10 November of the following year by storing egg masses at-2°C. L. monacha can be maintained continuously in the laboratory.

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Zusammenfassung Eine Massenvermehrung der Nonne, Lymantria monacha (L.), trat in 1984 auf 500 ha Pinus sylvestris bei Budelerbergen (S.O. der Niederlande) auf und betraf 1985 eine Fläche von 2800 ha. Obwohl die Vermehrung durch eine Luftapplikation mit Dimilin 25 bekämpft werden konnte, wurden Untersuchungen begonnen zur Entwicklung von für die Umwelt sicheren Bekämpfungsmethoden, besonders die Anwendung von Virosen. Das Ziel bestand in der Virusproduktion in künstlichen Raupenzuchten. Die Nonne wurde bisher mit verschiedenen für andere Insektenarten oder speziell für diesen Zweck entwickelten künstlichen Nährböden gezüchtet. Da die Zusammensetzung dieser Nährböden und die Zuchtmethoden nicht veröffentlicht waren und da wenige Details über die Raupenentwicklung vorlagen, wurden modifizierten Nährböden von McMorran sowie von Odell & Rollinson für die Massenzucht der Nonne geprüft.

     

Heritability estimates of phenthoate resistance in the diamond-back moth

Realized heritabilities were estimated for the character of phenthoate resistance in two local strains of the diamond-back moth, Plutella xylostella L. (Lepidoptera: Yponomeutidae), by performing artificial laboratory selection for resistance and susceptibility to phenthoate. Heritability estimates indicated that such traits are moderately heritable ( ²=0.42 and 0.41 in the resistant selection and ²=0.31 and 0.21 in the susceptible selection), and give an experimental basis accounting for rapid evolutionary changes of phenthoate resistance observed in field populations of this insect.The observed changes in variances of phenthoate susceptibility are discussed in relation to the additive genetic variance eliminated by directional selection. The explanation stresses the importance on the underlying genetic system.

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Résumé L'héritabilité de la résistance au phenthoate obtenue chez deux souches locales de P. xylostella L. (Lep.; Yponomeutidae) a été calculée en procédant au laboratoire à des sélections artificielles pour la résistance et pour la sensibilité à cet insecticide. Les calculs de l'héritabilité ont montré que de tels caractères sont moyennement héritables ( ²=0,42 et 0,41 lors de la sélection pour la résistance, et ²=0,31 et 0,21 lors de la sélection pour la sensibilité), et ont fourni la base expérimentale rendant compte des changements évolutifs rapides observés pour la résistance au phenthoate chez des populations naturelles de cet insecte.Les changements observés de la variance de la sensibilité au phenthoate sont discutés en fonction de la variance génétique additive éliminée par la sélection orientée. L'explication insiste sur l'importance du système génétique souligné.

     

Use of artificial eggs in studies of washout depth and drift distance for salmonid eggs

Colour-coded artificial trout eggs were used in investigations of washout depth in a natural stream and of drift distance relative to water velocity in an experimental channel and in a section of natural stream.Washout depth was studied in a spawning riffle of a stream whose bankful discharge is 5.6 m³ s^–1. During an experiment when spates never exceeded 6.5 m³ s^–1 egg washout was severe at 5 cm depth within the gravel, variable at 10 cm and negligible at 15 cm. During another experiment when a spate of 9.0 m³ s^–1 (return period 10–20 years) occurred, washout was severe at 5 and 10 cm depth and variable at 15 cm. There was also evidence that some eggs were moved short distances downstream within the gravel without being washed out.Within experimental channels, drift distance could be predicted from multiple regressions relating logarithms of water velocity, percentage of eggs settled and distance from point of release. At a water velocity of 100 cm s^–1 at 0.6 depth, 50% of eggs would settle within 8 m of the point of release. At water velocities of 75 to 100 cm s^–1 drifting eggs would, on average, travel at c. 60% of water velocity and make 1 to 2 bed contacts m^–1 of travel.A similar multiple regression can be applied to data from a natural stream channel. It predicts much larger drift distances (50% settled in 42 m at 100 cm s ^–1 ). However, in the natural channel, settlement appears aggregated and the validity of the concept of permanent settlement is in doubt.     

Artificial dimers of native actin: Preparation and properties in biological functions

With the aid of tartryl-bis--aminocaprylazide artificial dimers were produced from F actin from rabbit striated muscle. These derivatives will not polymerize by themselves but are able to copolymerize fully with native G actin. By modification of a single side chain per dimer, this copolymerization was completely inhibited. The dimers are able to activate subfragment I ATPase of myosin and bind to DNase I with inactivation of the enzyme in the same manner as native G actin. Within the dimer, one ADP is immobilized and will exchange against ATP extremely slowly. The dimers do not bind to the mushroom toxin phalloidin.