Detection of the EGFP sequence in breast muscle of 3‐year‐old chicken after transfection using sonoporation

In our continuing effort to generate transgenic chickens, sonoporation was chosen to insert an exogenous gene into the chicken genome. An EGFP expression vector (pCAG‐EGFPac) and microbubbles were injected into the central disc of stage‐X blastoderm or the germinal crescent of stage‐4 embryos, followed by ultrasonic vibration. Nineteen chicks out of 108 treated embryos hatched, six females and six males out of these 19 chicks grew to sexual maturity and two females and three males lived for 3 years. Genomic DNA from 17 out of 35 gonads from embryos and chicks that died before sexual maturity was EGFP‐positive by PCR. No EGFP sequence was detected in the genomic DNA of 322 embryos from six sexually mature females and the semen from four sexually mature males by PCR. When genomic DNA was obtained from various tissues of five 3‐year‐old chickens, the EGFP sequence was amplified from the genomic DNA of the breast muscle of a female (No. 85). The above sequence was subjected to DNA sequencing and verified to be the EGFP sequence. These results showed that sonoporation is an effective tool for the transduction of exogenous genes into chicken embryos for the generation of transgenic chickens.     

α-葡萄糖苷酶抑制剂的研究进展及食品源抑制剂的开发前景

随着世界上糖尿病患者的不断增加,除了传统的药物治疗以外,如今糖尿病及营养学专家更希望患者添加对辅助降血糖食品的应用,并建议用食疗的方法调节血糖水平,有效预防糖尿病的产生。在众多辅助降糖保健食品中,糖类物质、多酚类、黄酮类、磷脂酸类、生物碱等作为α-葡萄糖苷酶活性抑制剂,在降血糖方面起着极其重要的作用。     

DYNAMIC COMPUTED TOMOGRAPHY OF THE PANCREAS IN NORMAL DOGS AND IN A DOG WITH PANCREATIC INSULINOMA

To establish optimal imaging conditions for enhanced computed tomography (CT) for canine pancreatic tumors, 10 healthy beagles were subjected to dynamic CT. This technique was then applied to a dog with suspected insulinoma. The changes in mean peak enhancement and the delay time of the aorta and pancreas were determined. In normal beagles, maximal arterial and pancreatic CT enhancement was observed at 15 +/- 2 s (795 +/- 52 Housfield unit [HU]) and 28 +/- 9 s (118 +/- 16HU) after contrast medium injection, respectively. Multiphase enhanced CT was performed in a pug with suspected insulinoma using the CT protocol defined for the normal beagles with some parameters modified; the images were acquired at the arterial (14 s after contrast medium injection), pancreatic (after 28 s), and equilibrium (after 90 s) phases; scanning was followed by exploratory laparotomy. CT images were characterized by an enhanced mass in the left pancreatic lobe at the arterial phase, during which the difference between the CT values of the mass and normal pancreas was the highest. Histopathologic diagnosis of the pancreatic mass was insulinoma. Thus, it appears that enhanced CT imaging can be used to delineate the pancreas from a pancreatic mass, and it may be helpful in deciding the need for surgery.     

Horizontal distribution and seasonal variability of the epipelagic chaetognath Sagitta elegans in relation to hydrography in the western subarctic Pacific Ocean

The spatial distribution and seasonal variability of epipelagic chaetognaths along the Kurile Islands and off south-east Hokkaido, in the western subarctic Pacific Ocean. were investigated during the period from May 1990 to October 1992. Sagitta elegans was the dominant species among the epipelagic chaetognaths in the study area. Juvenile S. elegans were distributed mainly along the path of the mixed water which was determined by the acceleration potential anomaly (APA) at the iso-pycnal density surface of 26.6 σθ. The location of the path of The mixed water meandered around cyclonic and anticyclonic eddies. Catches of S. elegans were made at temperatures ranging from 4.5 to 22.2C. The juvenile distribution, however, was mostly restricted to between 1 and 4C of the potential temperature determined at the isopycnal density surface of 26.6 σθ. Variability in the juvenile abundance within the path of the mixed water could be caused by predation rather than by food limitation because microzooplankton, which are known to he prey for juveniles, were abundant enough to meet the food requirements of juveniles in the study area. Adults occurred in spring (April-May) and young individuals (juvenile and stage 1) were abundant in summer (June-July), when a strong thermocline developed. The main spawning period appears to be during April-May, with a possible second spawning period in the autumn.     

Variability in timing and magnitude of spring bloom in the Oyashio region, the western subarctic Pacific off Hokkaido, Japan

The spring bloom of phytoplankton is a well-established, regular, seasonal event in the western subarctic Pacific and is considered one of the most important conditions of massive production of pelagic fishes. A series of 12 cruises was conducted from 1990 to 1992 to examine the timing and magnitude of the spring phytoplankton bloom in the Oyashio region, the western subarctic Pacific off Hokkaido, Japan. An interannual variability in the bloom events was also analysed. On the basis of hydrographical characteristics, the study area was divided into three water masses: the Oyashio Water Mass, the Mixed Water Mass, and the Coastal Water Mass. Spring blooms were observed first in April in the Oyashio and the Coastal Water Masses, and continued to May in 1991 and 1992. However, no bloom was recorded in the Mixed Water Mass. High nutrient supply into the surface mixed layer during winter is likely to be one of the factors supporting an intense spring bloom in the Oyashio Water Mass. A significant positive relationship between log-transformed surface chlorophyll a concentration and maximum density gradient (MDG) within the euphotic layer was obtained in April, indicating the importance of vertical stability of the water column in the initiation of spring blooms in the Oyashio and the Coastal Water Masses. The spring blooms in 1991 were much more extensive and lasted longer than in 1990. It is suggested that meteorological conditions and abundance of grazers were responsible for this interannual difference.     

Signature-tagged mutagenesis of Vibrio vulnificus

Vibrio vulnificus is the causative agent of primary septicemia, wound infection and gastroenteritis in immunocompromised people. In this study, signature-tagged mutagenesis (STM) was applied to identify the virulence genes of V. vulnificus. Using STM, 6,480 mutants in total were constructed and divided into 81 sets (INPUT pools); each mutant in a set was assigned a different tag. Each INPUT pool was intraperitoneally injected into iron-overloaded mice, and in vivo surviving mutants were collected from blood samples from the heart (OUTPUT pools). From the genomic DNA of mixed INPUT or OUTPUT pools, digoxigenin-labeled DNA probes against the tagged region were prepared and used for dot hybridization. Thirty tentatively attenuated mutants, which were hybridized clearly with INPUT probes but barely with OUTPUT probes, were negatively selected. Lethal doses of 11 of the 30 mutants were reduced to more than 1/100; of these, the lethal doses of 2 were reduced to as low as 1/100,000. Transposon-inserted genes in the 11 attenuated mutants were those for IMP dehydrogenase, UDP-N-acetylglucosamine-2-epimerase, aspartokinase, phosphoribosylformylglycinamidine cyclo-ligase, malate Na (+) symporter and hypothetical protein. When mice were immunized with an attenuated mutant strain into which IMP dehydrogenase had been inserted with a transposon, they were protected against V. vulnificus infection. In this study, we demonstrated that the STM method can be used to search for the virulence genes of V. vulnificus.     

Expression of cell adhesion molecules in canine choroid plexus tumors

Choroid plexus tumor (CPT) is a primary intracranial neoplasm of the choroid plexus epithelium in the central nervous system. In the current World Health Organization classification, CPT is classified into two categories; choroid plexus papilloma (CPP) and carcinoma (CPC). In the present study, we investigated immunohistochemical expressions of N-cadherin, E-cadherin and β-catenin in 5 canine CPT cases (1 disseminated CPC, 2 CPCs and 2 CPPs). One CPP case was positive for N-cadherin and β-catenin, but negative for E-cadherin. The disseminated CPC case was positive for E-cadherin and β-catenin, but negative for N-cadherin. The other cases were positive for the three molecules examined. These results suggest that loss of the N-cadherin expression might associate with the spreading of CPC cells.     

Small‐scale heterogeneity in the soil environment influences the distribution of lawn grass and weeds

The association of the micro‐scale distribution of six plant species in a lawn, including the lawn grass, Zoysia japonica, with the soil's environmental factors was studied in an urban park in Kyoto, Japan. The cover of each plant species was scored in July and August. The relative elevation, depth of waterlogging after a shower, soil water content, and soil hardness were measured in 144 quadrats in a 12 m × 12 m plot. The subsurface soil was sampled at every fourth quadrat and was analyzed for the percentage of fine earth and the levels of total carbon, total nitrogen, and available phosphate. Three correlation criteria, namely the Pearson's, Mantel, and partial Mantel correlations, showed considerable disagreement in indicating whether or not there was a significant correlation between each plant species and environmental variable. Although the reason for the disagreement was not always clear, some unimodal or inverted unimodal responses of a plant species against an environmental factor, which was not detectable by the Pearson's correlation coefficient, were indicated to be significant by the Mantel or partial Mantel tests. There were four plant–environment pairs that had a significant correlation in all three criteria: Juncus tenuis in August had a positive correlation with the soil water content in July and August and a negative correlation with soil hardness and Z. japonica showed a negative correlation with the water content in August. The results suggest that the small patches of lawn with poor drainage are the sites of invasion by J. tenuis.     

Mid-to-caudal partial hemipelvectomy with limb preservation for ischial tumor in a dog

A 10-year-old spayed female Golden Retriever was referred for hindlimb lameness. A firm mass was palpated over the right caudal pelvis. Computed tomography revealed an osteolytic bone region and an associated periosteal reaction in the ischium, including the acetabulum. The histological diagnosis was sarcoma of unknown origin. A mid-to-caudal partial hemipelvectomy was performed to remove the mass. Femoral head and neck osteotomy was performed to allow hindlimb preservation. Following surgery, the dog regained satisfactory hindlimb use over time and was alive for 821 days with no recurrence or metastatic disease. This report indicates that mid-to-caudal partial hemipelvectomy with femoral head and neck osteotomy is technically feasible and allows for tumor control with preservation of the hindlimb and its function.