In vivo gene transfer into skeletal muscle of neonatal chicks by electroporation

Chicks (Gallus gallus domesticus) show considerable growth of skeletal muscle during the neonatal period. The in vivo gene transfer method is useful for studying gene function and can be employed to elucidate the molecular mechanisms of skeletal muscle growth in chicks. We evaluated the following conditions for gene transfer to the skeletal muscle of neonatal chicks by electroporation: (i) voltage; (ii) age of the chick; (iii) plasmid DNA injected amount; and (iv) duration of gene expression. The results obtained from this study indicate that the most efficient gene transfer condition was as follows: 75 µg of plasmid DNA encoding β‐galactosidase was injected into the gastrocnemius muscle of chicks at 4 days of age electroporated at 50 V/cm. In addition, peak transferred gene expression was observed from 3 days to 5 days after electroporation. Our results provide optimal electroporation conditions for elucidating the gene function related to skeletal muscle growth and development in neonatal chicks.     

DEVELOPMENT OF REFERENCE INTERVALS FOR THE LARGE INTESTINAL TRANSIT OF RADIOPAQUE MARKERS IN DOGS

Ten healthy dogs were fed 30 1.5 mm and 10 5 mm radiopaque markers (BIPS, MedID, Grand Rapids) mixed with sufficient quantities of a high fibre diet to meet 25% of their estimated daily caloric requirements. Abdominal radiographs were made at two hour intervals until 90% of the small and large markers had left the colon. The mean residence times (MRT) of each size of marker in the proximal, distal and total colon were calculated using kinetic analysis. The MRT's of the small markers were 4.9 hours (SD 4.4), 7.1 hours (SD 3.3) and 12.0 hours (SD 7.1) respectively. The MRT's of the large markers were not significantly different from the small markers except in the proximal colon where they were significantly shorter (3.2 hours, SD 2.3). Reference colonic filling and colonic transit curves for both sizes of markers were constructed. These may be useful to detect abnormal colonic transit in dogs.     

Rice <Emphasis Type="Italic">terpene synthase 18</Emphasis> (<Emphasis Type="Italic">OsTPS18</Emphasis>) encodes a sesquiterpene synthase that produces an antibacterial (<Emphasis Type="Italic">E</Emphasis>)-nerolidol against a bacterial pathogen of rice

Plant volatile compounds, including terpenes, are known to be involved in the rice defense system. In the present analysis of a terpene synthase, OsTPS18, in rice, we found that OsTPS18 was localized in the cytoplasm and synthesized the sesquiterpenes (E)-nerolidol and (E)-β-farnesene. The amounts of (E)-nerolidol and (E)-β-farnesene increased after jasmonic acid (JA) treatment. (E)-Nerolidol had significant antibacterial activity against Xanthomonas oryzae pv. oryzae (Xoo). These results suggest that (E)-nerolidol plays an important role in JA-induced resistance against Xoo and that it functions as an antibacterial compound in rice.     

Integrated Anaerobic-Aerobic Biodegradation of Multiple Contaminants Including Chlorinated Ethylenes,Benzene, Toluene,and Dichloromethane

Water, Air, & Soil Pollution - In the present research, the distribution and subcellular localization of cadmium in the roots, shoots, and leaves of Monochoria hastata were evaluated to...     

Purification and characterization of two anionic trypsins from the hepatopancreas of carp

SUMMARY: Two trypsins, designated as trypsin A and trypsin B, have been purified from the hepatopancreas of carp. The purification procedures consisted of ammonium sulfate fractionation, and chromatographies on DEAE-Sephacel, Ultrogel AcA54 and Q-Sepharose. Trypsin A was purified to homogeneity with the molecular mass of approximately 28 kDa, while trypsin B gave two close bands of 28.5 kDa and 28 kDa on sodium dodecylsulfate polyacrylamide gel electrophoresis both under reducing and non-reducing conditions. On native-PAGE, both trypsin A and trypsin B showed a single band. Trypsin A and trypsin B revealed optimum temperature of 40°C and 45°C, respectively, and shared the same optimum pH 9.0 using Boc-Phe-Ser-Arg-MCA as substrate. Both enzymes were effectively inhibited by trypsin inhibitors and their susceptibilities were similar. The NH₂-terminal amino acid sequences of trypsin A and trypsin B were determined to 37th and 40th amino acid residue, respectively. Their sequences were very homologous, but not identical to that of a trypsin-type serine proteinase from carp muscle and these of other trypsins. Immunoblotting test using the antibody raised against trypsin A cross-reacted with trypsin B positively.