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Optical problems, related to the particle on the surface, i.e. optical resonance and near-field effects in laser cleaning are discussed. It is shown that the small transparent particle with size by the order of the wavelength may work as a lens in the near-field region. This permits to focus laser radiation into the area with the sizes, smaller than the radiation wavelength. It leads to 3D effects in surface heating and thermal deformation, which influences the mechanisms of the particle removal.

Recently, the field of nanoelectonics has evolved into a major area of investigation. In this paper, we present a novel method of nanofabrication using pulsed lasers and near-field optical properties of sub-micron particles. For this purpose, spherical silica particles were deposited on a silicon surface. After laser illumination, hillocks with size of about 150 nm were obtained at the original position of the particles. The mechanism can be explained as the enhancement of light intensity near the contact area. Since the characteristic distance between particles and substrate is smaller than the radiation wavelength and the particle size is of the order of a wavelength, particle does not simply play the role of microfocusing lens as in far-field, but possess optical resonance effect in near-field. In our work, the light intensity on the surface under the spherical particle was calculated by solving the electromagnetic boundary problem “particle on suface”.

The synthesis of cholesterol esters by acyl-CoA:cholesterol acyltransferase (ACAT, EC 2.3.1.26) is an important component of cellular cholesterol homeostasis. Cholesterol ester formation also is hypothesized to be important in several physiologic processes, including intestinal cholesterol absorption, hepatic lipoprotein production, and macrophage foam cell formation in atherosclerotic lesions. Mouse tissue expression studies and the disruption of the mouse ACAT gene (Acact) have indicated that more than one ACAT exists in mammals and specifically that another enzyme is important in mouse liver and intestine. We now describe a second mammalian ACAT enzyme, designated ACAT-2, that is 44% identical to the first cloned mouse ACAT (henceforth designated ACAT-1). Infection of H5 insect cells with an ACAT-2 recombinant baculovirus resulted in expression of a approximately 46-kDa protein in cell membranes that was associated with high levels of cholesterol esterification activity. Both ACAT-1 and ACAT-2 also catalyzed the esterification of the 3beta-hydroxyl group of a variety of oxysterols. Cholesterol esterification activities for ACAT-1 and ACAT-2 exhibited different IC50 values when assayed in the presence of several ACAT-specific inhibitors, demonstrating that ACAT inhibitors can selectively target specific forms of ACAT. ACAT-2 was expressed primarily in mouse liver and small intestine, supporting the hypothesis that ACAT-2 contributes to cholesterol esterification in these tissues. The mouse ACAT-2 gene (Acact2) maps to chromosome 15 in a region containing a quantitative trait locus influencing plasma cholesterol levels. The identification and cloning of ACAT-2 will facilitate molecular approaches to understanding the role of ACAT enzymes in mammalian biology.

Southern blot analysis of mouse genomic DNA reveals two EcoRI fragments which faintly hybridize to mouse Adh-1 cDNA and are not part of the Adh-1 gene. These fragments were isolated from agarose gels, cloned, and characterized. Sequence analysis of the 2.1-kb EcoRI fragment suggests that it is likely a pseudogene since it does not contain a long open reading frame. However; the 2.0-kb EcoRI fragment contains a coding sequence with a long open reading frame which corresponds to exon 6 of the mouse Adh-1 gene. Comparison of the coding sequence with other known ADHs suggests that the sequence has diverged sufficiently from any currently known class of ADH to be a possible distinct class. Further confirmation awaits analysis of currently available genomic clones. Using these sequences as probe, restriction fragment length polymorphisms were identified for each sequence between C57Bl/6J and DBA/2J inbred mouse strains. The strain distribution pattern for these allelic differences was determined among the B x D recombinant inbred strains. This analysis revealed that the 2.1-kb EcoRI sequence is located on chromosome 3 but at a distance from the Adh-1/Adh-3 complex as previously reported. However, the new polymorphism identified in the 2.0-kb EcoRI fragment enabled this sequence to be mapped at the Adh-1/Adh-3 complex.

The hepatitis B virus X gene encodes a transcriptional trans-activator that appears essential for viral replication in the infected liver. It has been shown by others that at least two X protein products, initiating at different in-frame ATG codons within the X open reading frame, are necessary for full trans-activating function. Yet, previous studies have demonstrated only a full-length X gene transcript that extends 5' of the first ATG codon in the X open reading frame. In this communication, we present evidence for the existence of shorter X gene transcripts, whose heterogeneous 5' ends straddle the second ATG codon of the X open reading frame. These transcripts are not degradation products of the full-length X gene transcripts but rather arise from an independent promoter. Furthermore, immunofluorescence and functional analyses reveal that active X proteins are translated from these transcripts. Therefore, there appears to be a previously undescribed promoter embedded within the X open reading frame that can give rise to functionally important transcripts.

We present data demonstrating that hydrogen peroxide markedly decreases release of progeny hepatitis B virus particles in cultured cells. The presence of reduced glutathione prevents this effect. Hydrogen peroxide also decreases secretion of the hepatitis B virus surface and e antigens, with a concomitant decrease in the steady-state levels of the corresponding viral transcripts. This effect is specific to viral gene expression, since hydrogen peroxide at the concentration used does not have any significant effect on the overall pattern of host cell protein synthesis nor on the secretion of growth hormone from a co-transfected plasmid. Since hepatitis B virus can cause acute and chronic hepatitis and since inflammatory cells release significant amounts of reactive oxygen species, including hydrogen peroxide, this phenomenon may be of pathophysiological importance in the viral life cycle.

The molecular basis of the alcohol dehydrogenase (ADH)-negative deer mouse (Peromyscus maniculatus) has been investigated. Several classes of mammalian ADHs have been recognized based upon biochemical and structural properties. ADH cDNA clones identified by hybridization to a mouse class I ADH cDNA clone were obtained from a deer mouse ADH-positive liver cDNA library. This cDNA has been identified as being a class I sequence and represents the deer mouse Adh-1 gene. An additional cDNA sequence identified in both the ADH-positive and -negative deer mouse cDNA libraries was identified by weak cross-hybridization to the mouse cDNA. This cDNA encodes an amino acid sequence representing a new class of mammalian ADH, and the deer mouse gene for this ADH is named Adh-2. ADH-negative deer mice do not produce mRNA, that is detected by the Adh-1 cDNA probe. However, both stocks of deer mice produce high levels of Adh-2 mRNA in liver. Southern analysis using an essentially full-length Adh-1 cDNA probe has shown that the Adh-1 gene is deleted in the ADH-negative mice. Biochemical analysis of enzyme activity suggests at least three ADH polypeptides are expressed in different tissues and have somewhat different substrate specificities, as in the mouse.