Bacterial 16S ribosomal RNA genes (rDNA) were detected in blood samples from two healthy individuals by PCR under conditions involving 30 cycles that did not produce any visible products from negative control saline. Even from control samples, PCR involving 35-40 cycles yielded visible bands. Major clones detected in the blood samples, but not in control, were the Aquabacterium subgroup, Stenotrophomonas subgroup, Budvicia subgroup, Serratia subgroup, Bacillus subgroup and Flavobacteria subgroup. No clone was located within the bacteroides-clostridium-lactobacillus cluster, which is indigenous to gastrointestinal flora.Key words bacteremia, blood, polymerase chain reaction, 16S ribosomal RNA.Diagnosis of bacterial infection and identification of the agent responsible is essential in clinical medicine, and has been traditionally carried out by inoculating blood or infected tissues into a liquid or solid nutrient medium. This approach has a limitation in that it can detect only bacteria and fungi that are culturable in a laboratory. Recently, PCR using species-specific primers (1-5) and nucleic acid sequence-based amplification (NASBA) (6) have also been used as an alternative approach for detecting agents that are responsible for infection.These techniques can also be used for detailed analysis of microbial biota in the oral cavity and gastrointestinal tract, using universal primers that anneal to conserved regions in the 16S ribosomal RNA gene (rDNA) or gyr B gene (7-9). It is reported that half of the bacteria comprising the oral and intestinal flora have not been previously identified by in vitro culture procedures (10). Moreover, recent studies using PCR have raised the possibility that bacterial DNA may be present in the human bloodstream (11-13). Our ultimate objective is to elucidate the role of such subclinically infecting unculturable or latent bacteria in the pathogenesis of chronic vascular diseases (14-16). As a first step, we investigated whether bacteria can translocate in some way from the oral and intestinal flora to the blood stream in 'healthy' humans. In this preliminary study, blood specimen-specific bacterial sequences were detected by PCR of rDNA. However, they were not representative of sequences found in human intestine.PCR was done with a REDExtract-N-Amp Blood PCR kit (Sigma-Aldrich Japan, Tokyo, Japan) using broadrange 16S ribosomal RNA gene (rDNA)-specific oligonucleotide primers. The PCR kit does not require any type of purification, organic extraction, centrifugation or alcohol precipitation, and can be used with whole blood.For blood sampling, the skin was first sterilized with a cotton swab moistened with popidone iodide, the anterior brachial median vein was punctured using a sterile 21-gauge needle (Terumo Corporation, Tokyo, Japan),

An acenaphthylene-fused cyclo[8]pyrrole was synthesized by using an oxidative coupling reaction of the corresponding 2,2'-bipyrrole. Two conformational isomers 1 a and 1 b were isolated, and their molecular structures were elucidated by X-ray crystallographic studies. The less-polar and lower-symmetry 1 b isomer can be converted into the 1 a isomer through a thermal ring flip. Application of the perimeter model developed by Michl to magnetic circular dichroism spectroscopic data and theoretical calculations demonstrate that there is a marked redshift of the near-IR absorption maxima relative to cyclo[8]isoindole because there is a significant stabilization of the LUMO due to the differing effects of a fused ring expansion with acenaphthylene and benzene moieties on the frontier π molecular orbitals.

An acenaphthylene-fused cyclo[10]pyrrole 1b was selectively synthesized via an oxidative coupling reaction of the corresponding 2,2'-bipyrrole with the appropriate dianion template, croconate anion. The structure of 1b as the isolated largest cyclo[n]pyrrole was elucidated by X-ray crystallographic analysis. The absorption spectrum exhibited a markedly red-shifted, intensified L band at 1982 nm, which was interpreted by application of Michl's perimeter and Gouterman's 4-orbital models, supported by magnetic circular dichroism (MCD) data and theoretical calculations.

By a solid-state process, well-crystallized BaTiO3 (BT) particles with their average grain size below 0.2 μm were obtained. Wet and dry mechanical pretreatment processes were combined to obtain fine particulate mixture comprising BaCO3 and TiO2 with the highest possible homogeneity without causing appreciable agglomeration. Degree of homogenization was quantitatively evaluated by different microscopic techniques, in an attempt to optimize nuclei-growth processes. Reaction processes were discussed on the basis of thermal analyses in conjunction with the particulate morphology. The granulometrical and crystallographical properties of the present particulate products are comparable with or even superior to commercially available high-valued products fabricated via a hydrothermal or sol-gel route.

Retro: Cyclo[8]isoindole, which has a saddle‐shaped geometry (see picture, 2), was synthesized by oxidative coupling of a bicyclo[2.2.2]octadiene(BCOD)‐fused 2,2′‐bipyrrole followed by a retro‐Diels–Alder reaction of BCOD‐fused cyclo[8]pyrrole (1). Key trends in the optical spectra of ring‐annelated cyclo[8]pyrroles are identified based on magnetic circular dichroism spectra and theorectical calculations.