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.     

Sardine and anchovy regime fluctuations of abundance in four regions of the world oceans: a workshop report

Regimes of high abundance of sardine (Sardinops sagax and Sardina pilchardus) have alternated with regimes of high abundance of anchovy (Engraulis spp.) in each of the five regions of the world where these taxa co-occur and have been extensively fished. When one taxon has been plentiful, the other has usually been at a reduced level of abundance, and vice versa. Changes in the four heavily fished regions that support S. sagax–the Japanese, Californian, Humboldt, and Benguela systems–from a regime dominated by one taxon to a high level of abundance of the other have occurred more or less simultaneously. In the Pacific Ocean, sardines have tended to increase during periods of increasing global air and sea temperatures and anchovies to decrease. The Japanese system is dominated by sardines to a greater extent than the other systems, and sardines off Japan appear to increase as the Kuroshio Current cools. At the eastern edge of the Pacific Ocean, sardines colonize cooler areas during periods of warming. The Benguela system is out of phase with the three Pacific systems. The four systems all appeared to be in a state of flux in the 1980s. Increased abundance of the subdominant taxon is often one of the first signs of change. Sardines are relatively sedentary in refuge areas when scarce but change behavior to become highly migratory and colonize cooler areas when abundant. Anchovies, by contrast, expand around a fixed geographic center.     

Possible mechanisms underlying fluctuations in the Far Eastern sardine population inferred from time series of two biological traits

A cycle of fluctuation in the Far Eastern sardine, Sardinops sagax, population is accompanied by an abrupt drop and recovery of fish size and lipid content of parent fish, which may reflect a phase variation from coastal to oceanic and its reverse. The cycle begins with the survival success of a smaller number of eggs produced by high-quality parents, which is triggered by a global climate change (density-independent process), and finally ends in the survival failure resulting from an overexpansion of an enormous number of fish produced by low-quality parents (density-dependent process).     

Keratometry in normal cats: a cross-sectional study in Japan using an automated handheld keratometer

Keratometry was performed in 73 domestic cats of varied signalment in Japan using an automated handheld keratometer. The mean corneal curvature radius was significantly lower for cats younger than 1 year than for those older than 2 years (8.04 mm vs. 8.80−8.99 mm, P<0.01). The radius was significantly greater in males than in females among the cats older than 11 years (9.22 mm vs. 8.84 mm, P=0.01), while the age distributions of the males and females were similar. Corneal astigmatism did not significantly differ across the gender and age groups. The predictability of the corneal curvature and astigmatism was approximately 41−43% and less than 3%, respectively, as a function of age and bodyweight. The results highlighted some age- and sex-related keratometric variations in domestic cats in Japan.     

A new method for evaluating symplastic cadmium absorption in the roots of Solanum melongena using enriched isotopes 113Cd and 114Cd

Radioisotope techniques are well known as methods for evaluating symplastic ion absorption in roots. In the present study, a new method for evaluating symplastic cadmium (Cd) absorption in plant roots was developed using the enriched isotopes ¹¹³Cd and ¹¹⁴Cd. Seedlings of Solanum melongena were exposed to an enriched isotope solution of ¹¹³Cd at 25°C for 30 min. The roots were excised from each seedling and were then immersed in a cold buffer solution without Cd at 2°C for 120 min to suppress the metabolic activity of the roots. Finally, the roots were treated with a cold buffer solution containing enriched stable isotope ¹¹⁴Cd at 2°C for 120 min, whereby the apoplastically bound ¹¹³Cd was desorbed. We tested the validity of our method for evaluating symplastic Cd in roots compared with the conventional method based on differences in the amount of Cd absorbed at 2°C and 25°C using unlabeled Cd. There was no difference in the symplastic Cd content of the roots between the two methods. These results indicate that it is possible to evaluate the symplastic Cd content in roots using the enriched isotopes ¹¹³Cd and ¹¹⁴Cd.     

Changes in the milk composition of okapi (Okapia johnstoni) during the first 6 months of lactation

We collected 17 milk samples at 1 day to 6 months post partum from one captive okapi (Okapia johnstoni). Milks were examined for nutrient composition, including water, fat, crude protein, carbohydrates, ash and five minerals. The average values of the concentrations were 79.2 ± 3.0% SD water, 8.5 ± 3.4% fat, 7.5 ± 1.1% crude protein, 3.6 ± 0.6% carbohydrate, 1.2 ± 0.1% ash, 263.7 ± 47.3 mg/100 g calcium, 183.0 ± 40.0 mg/100 g phosphorus, 95.2 ± 17.6 mg/100 g sodium, 74.4 ± 34.3 mg/100 g potassium and 28.4 ± 4.4 mg/100 g magnesium, respectively. The protein concentration increased at late lactation (P < 0.01). The contents of sodium and potassium were rather high in the early lactation period, while those of calcium and phosphorus were rather low in the early lactation period. During the course of lactation in the first week post partum and the protein concentration was high at 1 day post partum. We compare our findings with the American Association of Zoos & Aquariums's Okapi Species Survival Plan Hand‐rearing Protocol, and recommend that nutrient composition of the formula for okapi calves is developed using the present study as a guide.     

MR IMAGING OF HISTIOCYTIC SARCOMA OF THE CANINE BRAIN

Histiocytic sarcomas are characterized by proliferation and/or infiltration of neoplastic histiocytes localized to specific organs, unlike malignant histiocytosis which involves many organ systems. Only a few cranial histiocytic sarcomas have been reported. Here we describe four dogs that presented with neurological deficits referable to the forebrain, and were diagnosed histologically as having histiocytic sarcoma. Using magnetic resonance (MR) imaging, the tumors were characterized by a T2-hyperintense and T1-isointense mass in one dog, T2- and T1-isointense extraaxial masses in two dogs, and a diffuse T2-hyperintense lesion over the left cerebral cortex in one dog. All tumors had contrast enhancement. MRI features in three of the four dogs were similar to that of meningioma, supported by the observation of a dural tail in two of these three dogs, and a broad base of attachment in the other. In the other dog the imaging findings were similar to those of encephalitis. Intracranial histiocytic sarcoma does not appear to have specific MR imaging features and can be confused with meningioma or encephalitis.     

Inactivation of Escherichia coli in soil by solarization

Contamination of agricultural soil by fecal pathogenic bacteria poses a potential risk of infection to humans. For the biosafety control of field soil, soil solarization in an upland field was examined to determine the efficiency of solarization on the inactivation of Escherichia coli inoculated into soil as a model microorganism for human pathogenic bacteria. Soil solarization, carried out by sprinkling water and covering the soil surface with thin plastic sheets, greatly increased the soil temperature. The daily average temperature of the solarized soil was 4–10°C higher than that of the non-solarized soil and fluctuated between 31 and 38°C. The daily highest temperature reached more than 40°C for 8 days in total in the solarized soil during the second and third weeks of the experiment. Escherichia coli in the solarized soil became undetectable (< 0.08 c.f.u. g⁻¹ dry soil) within 4 weeks as a result, whereas E. coli survived for more than 6 weeks in the non-solarized soil. Soil solarization, however, had little influence on the total direct count and total viable count of bacteria in the soil. These results indicate that soil solarization would be useful for the biosafety control of soil contaminated by human pathogens via immature compost or animal feces.