[Preparation and evaluation of in-situ synthesis zeolite gas-liquid modified columns].

A new kind of NaA zeolite column has been prepared by in-situ synthesis technique. In comparing with traditional coating method, the in-situ NaA zeolite carrier is much more uniform and rigid. Several kinds of zeolite gas-liquid modified columns have been prepared with this in-situ technology, which showed satisfactory features in separation property and thermal stability. The successful utilization of mini-length and wide-bore columns showed good practical prospect of this new kind of zeolite column.

Speckle in optical coherence tomography.

Speckle arises as a natural consequence of the limited spatial-frequency bandwidth of the interference signals measured in optical coherence tomography (OCT). In images of highly scattering biological tissues, speckle has a dual role as a source of noise and as a carrier of information about tissue microstructure. The first half of this paper provides an overview of the origin, statistical properties, and classification of speckle in OCT. The concepts of signal-carrying and signal-degrading speckle are defined in terms of the phase and amplitude disturbances of the sample beam. In the remaining half of the paper, four speckle-reduction methods-polarization diversity, spatial compounding, frequency compounding, and digital signal processing-are discussed and the potential effectiveness of each method is analyzed briefly with the aid of examples. Finally, remaining problems that merit further research are suggested. © 1999 Society of Photo-Optical Instrumentation Engineers.

Cross-polarized backscatter in optical coherence tomography of biological tissue.

We have observed that cross-polarized backscatter measured by optical coherence tomography of human skin in vivo is surprisingly strong. We identify and give evidence of its main origins: single scattering from nonspherical particles and multiple scattering by particles with sizes much larger than a wavelength. Our findings show that depolarized light scattered by dense large-diameter particles maintains a high degree of temporal coherence and that differential-polarization imaging improves contrast between particles of different sizes.

Molecular analyses of human immunodeficiency virus type 1 V3 region quasispecies derived from plasma and peripheral blood mononuclear cells of the first long-term-nonprogressing mother and child pair.

Molecular analyses were done for the V3 region quasispecies of human immunodeficiency virus type 1 (HIV-1) strains from plasma and peripheral blood mononuclear cells of the first HIV-1-infected long-term-nonprogressing mother-child pair whose members have survived for >13 years with stable CD4 T cell counts. There was a predominance of lower V3 loop charge and the absence of genotypic changes that are critical in phenotypic determination and tropism during HIV-1 infection. The intrahost genetic diversity between HIV-1 strains from the mother-child pair compared with HIV-1 strains from slow and rapid progressors suggested that a high genetic heterogeneity in HIV-1 strains from this HIV-1-infected long-term-nonprogressing mother and child pair was directly proportional to the length of their immunocompetent period.

Simian T cell leukemia virus type I from naturally infected feral monkeys from central and west Africa encodes a 91-amino acid p12 (ORF-I) protein as opposed to a 99-amino acid protein encoded by HTLV type I from humans.

A single protein of 12 kDa, p12 is encoded by the HTLV-I genome from both the singly spliced mRNA pX-ORF-I and doubly spliced mRNA pX-rex-ORF-I. While many full-length sequences of HTLV-1 are known, data on the p12 regions of African STLV-I are unavailable. We have undertaken to sequence the p12 gene in STLV-I from Central and West Africa naturally infected primates, and have compared them to known p12 sequences of HTLV-I. Our data on sequence and in vitro transcription-translation analyses indicate that p12 is a 91-amino acid (aa) protein among STLV-I strains from Central and West Africa, in contrast to the 99-aa protein found among HTLV-I strains around the globe. The p12 sequences of STLV-I exhibit a marked genetic variability at the level of both nucleotide and peptide sequences. Hydropathic and helical wheel analyses reveal that 60% of residues in HTLV-I p12 are hydrophobic, in contrast to 55% in STLV-I from Africa. Although HTLV-I and STLV-I show a similar putative antigenic site, a second potential site was located exclusively in STLV-I from Africa. There are differences in the predicted transmembrane domains in p12 between STLV-I and HTLV-I. Furthermore, the secondary structure data according to the Chou and Fasman algorithm predict an alpha-helical domain at the carboxy terminus in HTLV-I, and this domain may be truncated in STLV-I p12. The amino acid sequence of p12 shows two leucine zipper motifs (LZMs) at the amino terminus and in the middle region, respectively. This is the first report describing the size differences in p12 protein between HTLV-I and STLV-I, which may provide insights into pathogenic mechanisms used by HTLV-I and STLV-I.

Coinfection and genetic recombination between HIV-1 strains: possible biological implications in Australia and South East Asia.

It has been recognised that human immunodeficiency virus (HIV) mutates rapidly and that nucleotide substitutions, deletions, insertions, and rearrangements resulting from recombination events are the main factors that result in variation of the HIV-1 genome. Together, these processes are actively contributing to the diversity and virulence of viral forms comprising the acquired immune deficiency syndrome (AIDS) pandemic. There are 9 HIV-1 subtypes recognised (A-H and O), based on the envelope region segments. Inter-subtype recombination has been already described, whereas intra-subtype recombination has been difficult to detect. In this study, we have identified in vivo genetic recombination between HIV-1 strains belonging to subtype B in a patient who presented both intravenous drug use (IVDU) and homosexual sex as risk factors. Genetic analysis of viral strains in the hypervariable V3 region of the envelope gene indicated the presence of three distinct sequence groups categorized according to their respective tetrapeptide motifs-GPGR, GLGR and GPGK. Detailed genetic and phylogenetic analyses suggested the recombination occurring only between sequence groups with GPGR and GPGK tetrapeptide motifs. These data suggest that coinfection with closely related strains can occur in vivo, and the generation of hybrid HIV-1 genomes via genetic recombination between subtype B strains can result in further antigenic diversity which may thwart diagnosis and future vaccine efforts. Since HIV-1 subtype B is still the most commonly found subtype around the globe, the hybrid genomes between different subtype B strains may result in epidemiologic shifts and altered pathogenesis.

An HIV-1 infected long-term non-progressor (LTNP): molecular analysis of HIV-1 strains in the vpr and nef genes.

We describe a long-term non-progressive injecting drug user (IDU) who was infected with human immunodeficiency virus type-1 (HIV-1) in 1984, and has survived with stable CD4+ T-cell counts (> 800/microliters blood) without any acquired immune deficiency syndrome (AIDS) related illness. With a goal to investigate the molecular nature of HIV-1 strains infecting this patient, we amplified the nef and vpr genes directly from the fresh uncultured peripheral blood mononuclear cells (PBMCs), and carried out co-culture studies. Sequence analysis of the nef gene (from 1994 samples) showed no deletions (as has been previously reported) expected for a 7 base pair duplication at the C-terminus which prematurely terminated the nef reading frame, whereas even after repeated attempts the nef gene could not be amplified from the 1992 PBMC samples. In contrast, the vpr gene (from 1992 and 1994 samples) revealed two distinct quasispecies with no apparent defects. We observed five amino acid substitutions, between residues 83-90, at the C-terminus which has been recently implicated in G2 cell cycle arrest as an early step to HIV-1 infection. In the light of recent evidence on the role of nef gene defects/attenuations in long-term survival of HIV-1 infected patients, it may be that the nef gene defect created by gene duplication, which eliminated the cysteine-206 crucial in disulfide bond formation, may play a role in chronic HIV-1 infection in this patient. These data further suggest that deletions in the nef gene may not be the only reason for long-term non-progression of HIV-1 infection in some individuals, but the gene defects like duplication and subtle mutations in the functional motifs of both nef and vpr genes may confer similar protection in HIV-1 infected patients surviving for longer periods of time with stable CD4 counts.

Gene defects clustered at the C-terminus of the vpr gene of HIV-1 in long-term nonprogressing mother and child pair: in vivo evolution of vpr quasispecies in blood and plasma.

Earlier studies on HIV-1 strains from HIV-1-infected long-term nonprogressors (LTNP) have reported that nef deletions and/or attenuations may be crucial in the survival of these patients. Other reports have suggested that the nef gene may not be the only gene involved, but attenuations in other accessory genes (vif, vpr, vpu), which play an important role in the viral life cycle, may be similarly important in chronic HIV-1 infection in LTNPs. Here we show the molecular and phylogenetic analyses of the vpr gene in HIV-1 strains derived from both blood and plasma of an HIV-1 infected long-surviving mother-child pair which has survived for > 13 years with HIV infection: both have maintained stable CD4+ T-cell counts. Analyses of blood-and plasma-derived HIV-1 vpr clones indicated the presence of defects (insertions and deletions) and length polymorphisms. Interestingly, all the vpr defects in PBMCs and plasma were clustered at the C-terminus of the Vpr protein, between amino acid residues 83 and 89, which has been implicated in the G2 cell cycle arrest as a step to early HIV-1 infection. In contrast, the vpr sequence analysis of HIV-1 strains derived from 30 different patients, who either died of AIDS-related illnesses or have AIDS, showed neither C-terminal defects nor length polymorphism in the vpr gene. Also, secondary structure predictions suggest that the naturally occurring mutations at the C-terminal region (aa 83-89) have the potential to affect the secondary structure of the Vpr protein. Also, in some cases, the out-of-frame mutations and the length polymorphisms affect the tat gene reading frame. Together, these mutations may have potential significance in conferring chronic HIV-1 infection in this long-surviving nonprogressing mother-child pair.

Molecular analysis of the rfb gene cluster of a group D2 Salmonella enterica strain: evidence for its origin from an insertion sequence-mediated recombination event between group E and D1 strains.

The Salmonella enterica O antigen is a highly variable surface polysaccharide composed of a repeated oligosaccharide (the O unit). The O unit produced by serogroup D2 has structural features in common with those of groups D1 and E1, and hybridization studies had previously suggested that the D2 rfb gene cluster responsible for O-unit biosynthesis is indeed a hybrid of the two. In this study, the rfb gene cluster was cloned from a group D2 strain of S. enterica sv. Strasbourg. Mapping, hybridization, and DNA sequencing showed that the organization of the D2 rfb genes is similar to that of group D1, with the alpha-mannosyl transferase gene rfbU replaced by rfbO, the E1-specific beta-mannosyl transferase gene. The E1-specific polymerase gene (rfc) has also been acquired. Interestingly, the D1-like and E1-like rfb regions are separated by an additional sequence closely related to an element (Hinc repeat [H-rpt]) associated with the Rhs loci of Escherichia coli. The H-rpt resembles an insertion sequence and possibly mediated the intraspecific recombination events which produced the group D2 rfb gene organization.

Variation of the rfb gene clusters in Salmonella enterica.

In order to explore the genetic variation of O antigens of Salmonella enterica, we surveyed 164 strains (132 serovars) belonging to 45 serogroups, using 25 mostly single-gene rfb DNA probes for colony hybridization. The results revealed that strains within a serogroup have very similar or identical rfb genes. At least three of the four rhamnose genes were detected in all 17 serogroups reported to contain rhamnose, and one or more were detected in three others. The likelihood of being detected decreased in the order rfbB, rfbC, rfbA, and rfbD, which is the map order, suggesting a gradient of divergence. Mannose pathway genes were much less conserved, and of 27 groups reported to contain mannose or mannose derivatives colitose or fucose, only 9 hybridized to the rfbM and rfbK probes. Dideoxyhexose genes were found only in groups reported to contain dideoxyhexoses. Group D2, which had not been studied previously, appears to resemble group D1, with the substitution of one gene from group E1 to give a change in one linkage. In contrast to sugar pathway genes, sugar transferase genes did not in general hybridize to strains of other groups outside the closely related groups A, B, and D, with the exception of the galactose transferase gene also shared by groups C2, C3, and all E groups.