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     

Eye size variation reflects habitat and daily activity patterns in colubrid snakes

The functioning of the vertebrate eye depends on its absolute size, which is presumably adapted to specific needs. Eye size variation in lidless and spectacled colubrid snakes was investigated, including 839 specimens belonging to 49 genera, 66 species and subspecies. Variations of adult eye diameters (EDs) in both absolute and relative terms between species were correlated with parameters reflecting behavioral ecology. In absolute terms, eye of arboreal species was larger than in terrestrial and semiaquatic species. For diurnal species, EDs of terrestrial species do not differ from semiaquatic species; for nocturnal species the ED of terrestrial species is larger than fossorial species but not different from semiaquatic species. In relative terms, ED did not differ significantly by habitat for diurnal species. Although the ED of terrestrial species is larger than fossorial species there were no differences for nocturnal species between semiaquatic and fossorial snakes. In contrast to other vertebrates studied to date, colubrid EDs in absolute and relative terms are larger in diurnal than in nocturnal species. These observations suggest that among colubrid snakes, eye size variation reflects adaptation to specific habitats, foraging strategies and daily activities, independently of phylogeny. J. Morphol. 2012. © 2012 Wiley Periodicals, Inc.     

Isolation of microsatellite loci in the Capricorn silvereye,Zosterops lateralis chlorocephalus (Aves: Zosteropidae)

The Capricorn silvereye (Zosterops lateralis chlorocephalus) is ideally suited to investigating the genetic basis of body size evolution. We have isolated and characterized a set of microsatellite markers for this species. Seven out of 11 loci were polymorphic. The number of alleles detected ranged from two to five and observed heterozygosities between 0.12 and 0.67. One locus, ZL49, was found to be sex‐linked. This moderate level of diversity is consistent with that expected in an isolated, island population.     

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.     

Nervous system development in the fairy shrimp Branchinella sp. (Crustacea: Branchiopoda: Anostraca): Insights into the development and evolution of the branchiopod brain and its sensory organs

Using immunohistochemical labeling against acetylated a‐tubulin and serotonin in combination with confocal laser scanning microscopy and 3D‐reconstruction, we investigated the temporary freshwater pond inhabitant Branchinella sp. (Crustacea: Branchiopoda: Anostraca) for the first time to provide detailed data on the development of the anostracan nervous system. Protocerebral sense organs such as the nauplius eye and frontal filament organs are present as early as the hatching stage L0. In the postnaupliar region, two terminal pioneer neurons grow from posterior to anterior to connect the mandibular neuromeres. The first protocerebral neuropil to emerge is not part of the central complex but represents the median neuropil, and begins to develop from L0+ onwards. In stage L3, the first evidence of developing compound eyes is visible. This is followed by the formation of the visual neuropils and the neuropils of the central complex in the protocerebrum. From the deutocerebral lobes, the projecting neuron tract proceeds to both sides of the lateral protocerebrum, forming a chiasma just behind the central body. In the postnaupliar region, the peripheral nervous system, commissures and connectives develop along an anterior–posterior gradient after the fasciculation of the terminal pioneer neurons with the mandibular neuromere. The peripheral nervous system in the thoracic segments consists of two longitudinal neurite bundles on each side which connect the intersegmental nerves, together with the ventral nervous system forming an orthogon‐like network. Here, we discuss, among other things, the evidence of a fourth nauplius eye nerve and decussating projecting neuron tract found in Branchinella sp., and provide arguments to support our view that the crustacean frontal filament (organ) and onychophoran primary antenna are homologous. J. Morphol. 277:1423–1446, 2016. © 2016 Wiley Periodicals, Inc.     

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’.