Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

The importance of mother–infant communication for social bond formation in mammals

Mother–infant bonding is a universal relationship of all mammalian species. Here, we describe the role of reciprocal communication between mother and infant in the formation of bonding for several mammalian species. Mother–infant bond formation is reinforced by various social cues or stimuli, including communicative signals, such as odor and vocalizations, or tactile stimuli. The mother also develops cross‐modal sensory recognition of the infant, during bond formation. Many studies have indicated that the oxytocin neural system plays a pivotal role in bond formation by the mother; however, the underlying neural mechanisms for infants have not yet been clarified. The comparative understanding of cognitive functions of mother and infants may help us understand the biological significance of mother–infant communication in mammalian species.     

Dietary animal proteins alter monoamine metabolism in the brain

Several amino acids have effects on mental function, including sedative, antidepressant‐like and anxiolytic‐like effects. However, the influence of integrated amino acid nutrition as protein constituents on mental function remains unclear. Therefore, the purpose of the present study was to compare the influence of chicken, pork and beef protein extracts on brain monoamine metabolism in mice. Changes in monoamine levels and their turnover rates in the brain were induced by different protein sources. In particular, chicken protein group showed the highest norepinephrine levels in the hippocampus and hypothalamus, and beef protein extract caused an activation of the serotonergic system in the hypothalamus, although there were no significant differences in amino acid compositions of these protein extracts. Therefore, it was revealed that amino acid compositions in dietary protein did not induce alteration in monoamine metabolism. However, there were differences in small molecular peptides, such as creatine, carnosine and anserine levels in animal protein extracts. In conclusion, monoamine metabolism was altered by dietary protein sources. However, it was indicated that the alteration in monoamine metabolism may be independent from amino acid compositions in dietary protein. In addition, alteration in monoamine metabolism depending on the dietary protein sources may be induced by small molecular peptides.     

Impacts of acute imipramine treatment on plasma and brain amino acid metabolism in mice given graded levels of dietary chicken protein

Several studies have shown a relationship between depression and animal protein intake. To evaluate whether the difference of dietary chicken protein levels induces an antidepressant‐like effect and potentiates acute antidepressant effects, three levels of dietary chicken protein were used as the representative animal protein with imipramine used as the antidepressant. In addition, the effects of dietary chicken protein on brain metabolism were evaluated. Open field test (OFT) and forced swimming test (FST) were conducted on the 27th and 28th days, respectively. OFT and FST were not influenced by both imipramine and dietary protein levels. However, characteristic effects of imipramine treatment on brain monoamine metabolism were observed in the cerebral cortex and hypothalamus. In addition, dietary protein significantly increased taurine and L‐ornithine levels even though these amino acids were not contained in the diets. In conclusion, the metabolism of several amino acids in the plasma and brain were altered by dietary chicken protein.     

Validation of a newly generated oxytocin antibody for enzyme-linked immunosorbent assays

The biological and psychological significance of oxytocin is increasingly recognized; however, reliable assays of oxytocin in biological samples have not been developed. We raised a new oxytocin polyclonal rabbit antibody against synthetic oxytocin. The affinity of antibodies to oxytocin was examined by a radio-immunoassay and compared with that of a previously validated antibody. One antibody showed affinity for oxytocin in the radio-immunoassay. We developed a solid-phase ELISA for oxytocin using this antibody and compared it with existing methods. The newly developed ELISA showed comparable results using urine samples but not using serum samples. These results indicate that the new ELISA is useful for urinary oxytocin; further modifications, such as different extraction methods, are needed for its application to serum oxytocin.     

Predator–prey systems of drifting seaweed communities offthe Tohoku coast, northern Japan, as determined by feeding habitanalysis of phytal animals

ABSTRACT:   We collected phytal animals (fauna associated with drifting seaweeds)off the Tohoku coast in northern Japan in May and June 1994 to examine their feeding habits. Phytal animals were assignedto the following three feeding habit groups: (i) carnivores preyingon crustaceans: a gammarid ( Stenothoe sp.) and a decapodmegalops; (ii) omnivores feeding on crustaceans and pinnate diatoms:a cirripede ( Lepas anserifera ), three gammarids ( Calliopius sp., Jassaslatteryi and J. marmorata ), an isopod ( Idotea metallica ),a decapod crustacean ( Planes cyaneus ), a decapod megalopsand two caprellids ( Caprella penantis and Caprella mutica );and (iii) herbivores grazing on pinnate diatoms, fucoids and seaweedsother than fucoids: three gammarids ( Allorchestes angusta , Amphithoelacertosa and Perampithoe lindbergi ), an isopod ( Cymodocejaponica ) and a polychaete ( Platynereis bicanaliculata ).Pinnate diatoms in the gut contents were composed mainly of Licmophora spp., Navicula spp., Nitzschia spp.and Cocconeis spp. and seaweeds other than fucoids (i.e. Ectocarpussiliculosus and Sphacelaria furcigera ). The latter twoalgae seemed to be epiphytes on drifting seaweeds. These resultssuggest that phytal animals play an important role in drifting seaweed communitiesthrough trophic linkage with two primary producers (i.e. driftingseaweeds and the epiphytes growing on them).     

Effects of juvenile size at release and early marine growth on adult return rates for Hokkaido chum salmon (Oncorhynchus keta) in relation to sea surface temperature

Using path analyses, we investigated relationships between size at release from hatcheries, the early marine growth of juveniles, and adult return rates for chum salmon from five river stocks of Hokkaido, Japan, in relation to sea surface temperature during ocean residence. Marine growth was estimated using scales collected from 11 760 adults of age 0.3 (1980–2004). The growth and survival of each stock appeared to have a different suite of regulatory processes. Interannual variability in return rates was mainly regulated by size at release in two stocks from the Sea of Okhotsk. A similar relationship was found in one stock from the Sea of Japan, but growth during coastal residency also affected their return rates. In two stocks from the Pacific coast of Hokkaido, variability in return rates was not related to size at release or to the coastal growth of juveniles, but with offshore growth in the Sea of Okhotsk, the nursery area for juveniles after leaving Japanese coastal waters. Whereas coastal growth tended to be negatively correlated with size at release in some stocks, offshore growth was positively associated with the August–November sea surface temperature in all stocks. This study confirmed that mortality of juvenile salmon occurred in two phases, during the coastal residency and the late period of the growing season, but the relative importance of both phases varied by stock and region, which probably regulated year‐class strength of Hokkaido chum salmon.     

Phylogenetic characterization of Isospora jaracimrmani oocysts from a veiled chameleon (family Chamaeleonidae; Chamaeleo calyptratus) reared at a zoo in Ishikawa,Japan

Oocysts of Isospora sp. were detected in the feces of a veiled chameleon (family Chamaeleonidae; Chamaeleo calyptratus) kept at a zoo in Ishikawa, Japan. Phylogenetic analysis placed the sequence in the cluster of Isospora spp. isolated from reptiles. Based on a comparison of morphological data of ten previously reported Isospora species from the Chamaeleonidae family, this isolate was morphologically similar to I. jaracimrmani, which has been considered to be a virulent species. This case study suggests the possibility that species of Isospora might not always cause disease because the animal that shed these oocysts showed no symptoms for more than two months.     

Evaluation of a rapid coliform detection kit from clinical mastitis milk using colloidal gold nanoparticle–based immunochromatographic strips

The accurate identification of mastitis‐causing bacteria assists in effective management by both dairy farmers and veterinarians and can be used to implement the selective use of antimicrobials for treatment. The purpose of this study was to evaluate the ability of our developed anti–ribosomal protein-L7/L12 antibody–coated immunochromatographic strip (ICS) test to detect coliforms in milk by comparing the results with the bacteriological culture method. We investigated the performance of the ICS test as compared with the bacteriological culture method using 308 milk samples from clinical bovine mastitis. First, to determine the optimal ICS test cutoff point for detecting coliform mastitis, we developed a receiver-operating characteristic curve. The result showed that the cutoff point was at 0.5 of our index. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value of the ICS test were 81.3%, 84.8%, 69.2%, and 91.54%, respectively. As the clinical signs increased in severity, the F-measure, a weighted harmonic mean of the sensitivity and overall PPV performance, increased. Because it is especially important to treat clinical mastitis appropriately in the early stages of detection, the ICS test, which can be used by both dairy farmers and veterinarians on dairy farms, is considered to be a useful tool for detecting coliform mastitis, which often presents with severe signs.     

Effects of starvation on RNA : DNA ratio, glycogen content, and C : N ratio in columellar muscle of the Japanese turban shell Turbo (Batillus) cornutus (Gastropoda)

ABSTRACT: The validity of using the RNA : DNA ratio, glycogen content, and C : N ratio in the columellar muscle as indicators of the nutritional condition in the Japanese turban shell Turbo cornutus was examined. These biochemical indices were compared between fed and starved animals. The fed animals were fed brown algae to satiation for 108 days as a control, whereas the starved animals were not fed for the first 75 days and then fed for next 33 days. All three indices declined during the starvation period, and the values for the starved group were significantly lower than those for the fed group during days 20–75. At day 108, after the re-feeding period, the indices for the starved group were found to have increased. Among the indices, only the RNA : DNA ratio for the starved males responded rapidly to starved conditions and became significantly lower even at day 4, and only the RNA : DNA ratio for the starved males and females recovered to those levels of the fed animals at day 108. The results indicated that the RNA : DNA ratio is the most rapid indicator of nutritional stress among the three indices. However, it is recommended that glycogen content and C : N ratio be used in addition to the RNA : DNA ratio for monitoring the health of T. cornutus as the RNA : DNA ratio showed large variations.