The distribution of serum C-reactive protein (CRP) levels and their association with age, sex, and atherosclerotic risk factors were studied in a large Japanese population between 1992 and 1995. The subjects consisted of 2,275 males and 3,832 females aged 30 years and over. CRP was measured by nephelometry. The distribution of CRP was highly skewed toward a lower level than that of previous studies and seemed to be a combination of two separate distribution curves. The increase in CRP with age was statistically significant, and males had higher CRP levels than did females. Males who were current smokers had higher CRP levels than did nonsmokers. Age, systolic blood pressure, diastolic blood pressure, triglycerides, fibrinogen, and body mass index were all positively associated with CRP in both sexes, while total cholesterol and blood glucose were positively related in females only. High density lipoprotein cholesterol was inversely related in both sexes. Multiple logistic regression analysis showed that sex, age, systolic pressure, high density lipoprotein cholesterol, triglycerides, fibrinogen, and body mass index were significant independent variables. In conclusion, the distribution of CRP among the Japanese was quite different from that among Westerners, although CRP levels correlated with other atherosclerotic risk factors, similar to those in Westerners.

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),

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