Effects of atherosclerosis on mean and daily variation of arterial pressure in conscious WHHL rabbits.

Effects of atherosclerosis on the mean value and daily variation of arterial pressure were studied in 12 Watanabe-heritable hyperlipidemic (WHHL) rabbits aged 12 to 35 months and 25 normal Japanese white rabbits aged 6 to 30 months. A pressure catheter was inserted through the left subclavian artery under pentobarbital anesthesia. A few days after the catheterization, the mean arterial pressure (MAP) of the rabbits, which were active and in a good state of appetite, was recorded by an analogue-to-digital converter every second for about 6 hrs and stored in a computer. The mean (M) and standard deviation (SD) in the WHHL rabbit, calculated from each successive MAP record, ranged widely from 85.8 to 131.4 mmHg and 5.6 to 12.6 mmHg, respectively. There was no significant correlation between M and SD in the WHHL rabbit. M and variance (V) of MAP in the WHHL rabbit were significantly higher than those in the normal rabbit. M did not show any significant change with increasing ages, whereas SD increased significantly with aging in the WHHL rabbit. Concentrations of serum total cholesterol and triglyceride in the WHHL rabbit were 475 and 328 mg/dl, which were about nine and seven times as high as those in the normal rabbit, respectively. Macroscopic and histopathological examinations of the aorta revealed development and spread of sclerotic lesions with aging in the WHHL rabbit. We can conclude that development of atherosclerosis with aging in the WHHL rabbit causes malfunction of the baroreceptors, which contributes to hypertension and lability of arterial pressure.     

Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

Reproductive biology of the female Japanese mantis shrimp Oratosquilla oratoria (Stomatopoda) in relation to changes in the seasonal pattern of larval occurrence in Tokyo Bay, Japan

ABSTRACT:   The reproductive traits and the monthly larval abundance of the mantis shrimp Oratosquilla oratoria were investigated in Tokyo Bay, Japan, in 2002. The goal of the study was to elucidate the cause of changes in the monthly pattern of larval abundance from the 1980s to the 1990s as these changes relate to variation in the stock size of the adult shrimp. Oogenesis was divided into 10 stages by histological observation. The developmental stage of oocytes in an individual's ovary was synchronous, suggesting that almost all the oocytes in an ovary are spawned at the same time. The size at first maturity was estimated to be 7 ≤ body length ( BL ) < 8 cm. Fecundity was expressed as a function of BL , ranging from 19 300 eggs for 8 cm BL to 92 100 eggs for 14 cm BL . Small female shrimps (<10 cm BL ) spawned around August. Most large female shrimps (≥10 cm BL ) spawned around May, and some large female shrimps also spawned until September. Although most large female shrimps spawned in spring, the larval abundance was low before July and high from August onwards. The results suggest that a substantial decrease in the stock size of large individuals causes the low larval abundance before July.     

Pilot Production of Hatchery-Reared Summer Flounder Paralichthys dentatus in a Marine Recirculating Aquaculture System: The Effects of Ration Level on Growth, Feed Conversion, and Survival

Pilot‐scale trials were conducted to evaluate growout performance of hatchery‐reared summer flounder fingerlings in a state‐of‐the‐art recirculating aquaculture system (RAS). The outdoor RAS consisted of four 4.57‐m dia × 0.69‐m deep (vol. =11.3 m³) covered, insulated tanks and associated water treatment components. Fingerlings (85.1 g mean initial weight) supplied by a commercial hatchery were stocked into two tanks at a density of 1,014 fish/tank (7.63 kg/m³). Fish were fed an extruded dry floating diet consisting of 50% protein and 12% lipid. The temperature was maintained between 20 C and 23 C and the salinity was 34 ppt. Under these conditions, growth, growth variation (CV_wt), feed utilization, and survival of fish fed to 100% and 82% of a satiation rate were compared. Due to clear changes in growth patterns during the study, data was analyzed in three phases. During phase 1 (d 1–d 196), fish showed rapid growth, reaching a mean weight of 288 g ± 105 and 316 g ± 102, with a CV_wt of 0.36 and 0.32 and FCR's of 1.38 and 1.36 in the subsatiation and satiation groups, respectively. During phase 2 (d 196–d 454), fish displayed slower growth reaching mean weights of 392 g ± 144 and 436 g ± 121, with a CV_wt of 0.37 and 0.28, and increasing FCR's of 3.45 and 3.12 in the subsatiation and satiation groups, respectively. During phase 3 (d 454–d 614), fish showed little growth reaching mean weights of 399 g ± 153 and 440 g ± 129, with a CV_wt of 0.38 and 0.29 in the subsatiation and satiation groups, respectively. Over the entire growout period (d 1–d 614), feed conversion ratios were 2.39 and 2.37 and survival was 75% and 81 % in the subsatiation and satiation treatments, respectively. The maximum biomass density reached during the study was 32.3 kg/m³. The satiation feed rate was superior to the 82% satiation rate, since it maximized growth rates, with no effect on FCR. The higher CV_wt in the subsatiation group suggests increased competition for a restricted ration led to a slower growth with more growth variation. The decrease in growth in phases 2 and 3 was probably related to a high percentage of slower growing male fish in the population and the onset of sexual maturity. This study demonstrated that under commercial scale conditions, summer flounder can be successfully grown to a marketable size in a recirculating aquaculture system. Based on these results, it is recommended that a farmer feed at a satiation rate to minimize growout time. More research is needed to maintain high growth rates through marketable sizes through all‐female production and/or inhibition of sexual maturity.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.