DNA transfection using linear poly(ethylenimine) prepared by controlled acid hydrolysis of poly(2-ethyl-2-oxazoline).

A series of linear poly(ethylenimine) (L-PEI) containing varying amounts of cationic charge density in its backbone was produced by controlled hydrolysis of poly(2-ethyl-2-oxazoline) (PEtOz) for using as a nonviral DNA transfection agent. The effects of cationic charge density and molecular weight of the L-PEI on the cytotoxicity and transfection efficiency were studied. The efficiency of transfection was monitored by using a luciferase reporter gene system. Gel retardation assay and dynamic light scattering (DLS) showed that the condensation capacity of L-PEI was suitable for transfection. Highly compacted L-PEI/DNA complex ( approximately 150 nm) was obtained with a surface charge value of around +28.4 mV. Cell cytotoxicity was affected to a great extent by the hydrolysis percent of L-PEI as well as by the molecular weight. Transfection efficiency of luciferase plasmid DNA against NIH 3T3 fibroblast was largely dependent upon the hydrolysis percent (charge density) in the polymer backbone and the molecular weight of the L-PEI, but independent of the total amount of cationic charges used for DNA condensation. L-PEI with a hydrolysis percent of 88.0% exhibited comparable transfection efficiency to that of commonly used branched PEI.

Poly(DMAEMA-NVP)-b-PEG-galactose as gene delivery vector for hepatocytes.

A block copolymer composed of cationic polymer and poly(ethylene glycol) (PEG) was used as a DNA carrier. Poly(2-(dimethylamino)ethyl methacrylate (DMAEMA)-co-N-vinyl-2-pyrrolidone (NVP)) having a terminal carboxylic group was synthesized by free radical polymerization using an initiator, 4,4'-azobis(4-cyanovaleric acid). The terminal carboxylic acid was activated by N-hydroxysuccinimide (NHS) with dicyclohexylcarbodiimide (DCC) and then conjugated with PEG-bis(amine). For specific gene targeting to asialoglycoprotein receptor of hepatocytes, a galactose moiety was incorporated into the PEG terminal end of poly(DMAEMA-NVP)-b-PEG by reductive coupling using lactose and sodium cyanoborohydride. RSV luciferase plasmid was used as a reporter gene, and in vitro gene transfection efficiency was measured in HepG2 human hepatocarcinoma cells. Poly(DMAEMA-NVP)-b-PEG-galactose/DNA complexes formed at 0.5-2 polymer/plasmid weight ratio had compacted structures around 200 nm particle size and exhibited slightly negative surface charge. These complexes were coated with a cationic, pH sensitive, endosomolytic peptide, KALA, to generate positively charged poly(DMAEMA-NVP)-b-PEG-galactose/DNA/KALA complex particles. In the presence of serum proteins, both the PEG block and the galactose moiety of poly(DMAEMA-NVP)-b-PEG-galactose greatly enhanced the gene transfection efficiency, which was very close to that of Lipofectamine plus. Irrespective of the presence of serum proteins, as the KALA/DNA weight ratio increased, the transfection efficiency of poly(DMAEMA-NVP)-b-PEG-galactose was enhanced due to the pH dependent endosomal disruptive property of KALA. This study demonstrates that sufficient transfection efficiency as high as that of commercial agent could be attained by judicious formulation of molecular engineered poly(DMAEMA-NVP)-b-PEG-galactose in combination with an endosomolytic peptide, KALA.

NsiI and ScaI restriction fragment length polymorphisms at the atrial natriuretic peptides (ANP) gene locus.

The authors report on two new restriction fragment length polymorphisms at the human atrial natriuretic peptide gene locus, detected in three families with restriction endonucleases ScaI and NsiI.

Effects of the acoustic noise of the gradient systems on fMRI: a study on auditory, motor, and visual cortices.

MR acoustic, or sound, noise due to gradient pulsing has been one of the problems in MRI, both in patient scanning as well as in many areas of psychiatric and neuroscience research, such as brain fMRI. Especially in brain fMRI, sound noise is one of the serious noise sources that obscures the small signals obtainable from the subtle changes occurring in oxygenation status in the cortex and blood capillaries. Therefore, we have studied the effects of acoustic, or sound, noise arising in fMR imaging of the auditory, motor, and visual cortices. The results show that the effects of acoustic noise on motor and visual responses are opposite. That is, for motor activity, there is an increased total motor activation, whereas for visual stimulation, the corresponding (visual) cortical activity is diminished substantially when the subject is exposed to a loud acoustic sound. Although the current observations are preliminary and require more experimental confirmation, it seems that the observed acoustic-noise effects on brain functions, such as in the motor and visual cortices, are new observations and could have significant consequences in data observation and interpretation in future fMRI studies.

Nasal cycle in patients with septal deviation: evaluation by acoustic rhinometry.

The nasal cycle in patients with septal deviation was studied by acoustic rhinometric techniques. This study included 24 patients with anteriorly located septal deviations (mean age = 23.5), and 26 normal controls (mean age = 24.7). Data of MCA (minimum cross-sectional area) and NV (nasal volume), collected in 20-minute intervals, were plotted for each subject during 8 hours. Twenty of 24 patients (83%) with septal deviation and 20 of 26 normal subjects (77%) showed at least one complete cycle. Duration of the nasal cycle, which ranged from 100 minutes to 400 minutes, had no statistical difference between the septal deviation group (mean duration of 216 minutes) and the normal control group (mean duration of 227 minutes). The degrees of variation of MCA and NV, defined as Degree of Variation of MCA (%) = 100 (MCAmax - MCAmin)/MCAmax, Degree of Variation of NV (%) = 100 (NVmax - NVmin)/NVmax, which represent the percent change of MCA and NV throughout the study, showed no difference between the wide side and the narrow side, or between the septal deviation group and the normal control group. These findings suggest that the nasal cycle is relatively independent of peripheral anatomic factors for its generation. However, the amplitude of changes of MCA was greater in the wide side, and the sum of both MCAs tended to fluctuate in accordance with the fluctuation of MCA of the wide side. Thus, the nasal cycle seemed to be affected by septal deviation.

Normative measurements of Korean orbital structures revealed by computerized tomography.

PURPOSE: The purposes of this study were to establish criteria for the diameters of normal extraocular muscles, the width of the optic nerve, and the globe position as revealed by CT, and to investigate the effects of age and sex on these parameters in the Korean population. SUBJECTS AND METHODS: Diameters of extraocular muscles (medial, lateral, superior complex, and inferior rectus), distance from the interzygomatic line to the posterior margin of the globe, width of the optic nerve-sheath complex, and length of the interzygomatic line were calculated for 214 patients on axial and direct coronal CT images, and the effects of age and sex were analyzed. RESULTS: Normal range for the diameters as given by mean+/-2SDs of extraocular muscles were medial rectus, 2.2--5.4 mm; lateral rectus, 2.1--4.9 mm; inferior rectus, 2.5--5.7 mm; and superior complex group, 2.6--6.4 mm. The mean optic nerve-sheath complex was 4.1 mm and the values ranged from 2.9 mm to 5.3 mm. The mean length of the interzygomatic line was 103.8 mm. The normal posterior position of the globe as here designed was 11.2 mm behind the interzygomatic line (range, 6.4--15.3 mm). All parameters under study were a little larger in males than females. Statistically, however, there was no significant difference between males' and females' results, nor between the various age groups (p>0.05). CONCLUSION: Our CT results may help clinical ophthalmologists to accurately assess enlargement of the extraocular muscles and the optic nerve as well as aiding in the evaluation of exophthalmos.