Complement-mediated bacteriolysis after binding of specific antibodies to drug-resistant Pseudomonas aeruginosa: morphological changes observed by using a field emission scanning electron microscope.

A bactericidal mechanism mediated by human serum was investigated by a field emission scanning electron microscope and a strain of drug-resistant Pseudomonas aeruginosa. When the bacteria were treated with meropenem, a carbapenem antibiotic, spheroplasts and bulges (spheroidization) appeared after 1-3 h. When 40% serum was added to the bacteria, the bacteria agglutinated within 2 min and then lysed after 5-30 min. Immunoelectron micrographic analyses showed dispositions of complement component C9 molecules on the cell surface of lysed bacteria by the serum treatment that might suggest formation of a membrane attack complex. Immunoglobulin G (IgG) depletion from the serum diminished the lytic activity and adding human intravenous immunoglobulin (IVIG) restored it, suggesting that lysis was induced by specific IgG binding to the bacteria. IVIG may help patients with less IgG against bacteria to overcome severe infection.

Enzymatic detachment of spore germlings in Magnaporthe oryzae.

Magnaporthe oryzae germlings tightly attach to the host surface by producing extracellular matrix (ECM) from germ tubes and appressoria, which are important for the early infection process. To understand the adhesion mechanisms of ECM during differentiation of infection structure, we evaluated the effects of various enzymes on M. oryzae germlings and the disease symptoms of the host plant, wheat. Treatment with ß-mannosidase, collagenase N-2, collagenase S-1, or gelatinase B at 1-h postinoculation (hpi) resulted in germling detachment, although producing normal appressoria. Treatment with matrix metalloproteinases (MMPs) at 6 hpi also caused germling detachment. Furthermore, we confirmed by the inoculation tests and scanning electron microscopy that the germlings on the wheat plant were removed and did not manifest pathogenicity on treatment with MMPs. The most effective MMPs were crude collagenase, collagenase S-1, and gelatinase B, suggesting that the application of MMPs is promising for crop protection from fungal diseases by its detachment action.

Cellular Injury of Cardiomyocytes during Hepatocyte Growth Factor Gene Transfection with Ultrasound-Triggered Bubble Liposome Destruction.

We transfected naked HGF plasmid DNA into cultured cardiomyocytes using a sonoporation method consisting of ultrasound-triggered bubble liposome destruction. We examined the effects on transfection efficiency of three concentrations of bubble liposome (1 × 10(6), 1 × 10(7), 1 × 10(8)/mL), three concentrations of HGF DNA (60, 120, 180 µg/mL), two insonification times (30, 60 sec), and three incubation times (15, 60, 120 min). We found that low concentrations of bubble liposome and low concentrations of DNA provided the largest amount of the HGF protein expression by the sonoporated cardiomyocytes. Variation of insonification and incubation times did not affect the amount of product. Following insonification, cardiomyocytes showed cellular injury, as determined by a dye exclusion test. The extent of injury was most severe with the highest concentration of bubble liposome. In conclusion, there are some trade-offs between gene transfection efficiency and cellular injury using ultrasound-triggered bubble liposome destruction as a method for gene transfection.