Quantification of Blautia wexlerae and Blautia luti in human faeces by real-time PCR using specific primers.

The Clostridium coccoides group, including the genus Blautia and other genera, is one of the predominant bacterial groups in the human intestine. We re-examined 266 human faecal clones and 58 isolates in the C. coccoides group isolated by Hayashi et al. (2002) in order to elucidate the detailed distribution of Blautia wexlerae and Blautia luti in human faeces. Subsequently, we designed a primer pair specific for B. wexlerae and B. luti based on the 16S ribosomal RNA (16S rRNA) gene sequence. The number of B. wexlerae and B. luti in faecal samples of 12 healthy Japanese subjects was examined by real-time PCR assay. The number of the C. coccoides group in the 12 faecal samples was also determined using C. coccoides group-specific primers. Re-examination of the human faecal clones and isolates revealed that B. wexlerae and B. luti accounted for 19.5% of the clones and 25.9% of the isolates. B. wexlerae and B. luti were detected in all faecal samples with 5.3±3.2×10(9) cells/g faeces (wet weight, average ± standard deviation) as assessed by real-time PCR. Furthermore, B. wexlerae and B. luti constituted 32.3±12.7% (average ± standard deviation) of the C. coccoides group (1.7±0.8×10(10) cells/g faeces). This demonstrates that B. wexlerae and B. luti were presented in human faeces with a high frequency as the dominant bacteria.

Analysis of the human intestinal microbiota from 92 volunteers after ingestion of identical meals.

The intestinal microbiota composition of 92 volunteers living in Japan was identified following the consumption of 'identical meals' (1,879 kcal/day) for 3 days. When faecal samples were analysed by terminal restriction fragment length polymorphism with several primer-restriction enzyme systems and then clustered, the patterns could be divided into 2 clusters. Contribution tests and partition modelling showed that OTU211 of the 35f-MspI system and OTU237 of the 35f-AluI system were key factors in the distribution of these groups. However, significant differences among these groups in terms of body mass index and age were not observed.

[The latest trend of PRSP infection].

Streptococcus pneumoniae is an important pathogen and is a leading cause of pneumonia, meningitis, otitis media and sinusitis. Penicillin-resistant Streptococcus pneumoniae(PRSP) first appeared in the 1970s, and its has been increasing recently worldwide. Penicillin resistance among streptococcal isolates has been rapidly emerged in Japan the last decade, too. Penicillin and beta-lactam resistance due to altered penicillin binding protein was recognized. Resistant strains to other non-beta-lactam drugs, such as the macrolides, clindamycin, minocycline began to increase. Despite the use of appropriate antimicrobial agents, therapeutic failure in-patients with pneumococcal infection were increasing. It will be more important to select antimicrobial agent for PRSP infection, especially in meningitis, where the minimum inhibitory concentration of drug in cerebrospinal fluid may be less than the MIC in the blood. In the future 23-valent pneumococcal capsular polysaccharide vaccine should be used all persons 2 years of age and older who are at risk for serious pneumococcal disease and in all persons 65 years of age and older.

[Cerebral palsy in twins in Okinawa].

Twin pregnancy is an important risk factor associated with cerebral palsy (CP). We investigated the prevalence and clinical features of CP in twins born from 1988 to 1994 in Okinawa prefecture. The prevalence of CP in twins was 10.1 per 1000 multiple births, and it was 6 times that in singletons (1.6 per 1000 live births). Twins weighing more than 2500 g had a higher prevalence of CP than singletons, but those weighing less than 2500 g had no significant difference. Our results indicate that the higher rate of CP in twins reflect their tendency to be low-birth weight. There may be specific events affecting twins weighing 2500 g or more.

[Diffuse aspiration bronchiolitis in an elderly woman].

A 74-year-old woman was admitted to the hospital because of persistent nocturnal coughing and abnormal shadows on chest x-ray films. The films showed cavities in the right upper lobe and small nodular shadows in the right upper, lower, and left upper lung fields. Examination of a specimen obtained by transbronchial lung biopsy showed nonspecific inflammatory changes. An open-lung biopsy was done. Histopathological examination showed evidence of diffuse aspiration bronchiolitis and actinomyces. Actinomyces is a member of the endogenous flora of the oral mucous membranes. Our diagnosis was diffuse aspiration bronchiolitis caused by micro-aspiration of oro-pharyngeal secretions during sleep.

[Susceptibilities of bacteria isolated from patients with respiratory infectious diseases to antibiotics (1995)].

The bacteria isolated from the patients with lower respiratory tract infections were collected by institutions located throughout Japan, since 1981. Ikemoto et al. have been investigating susceptibilities of these isolates to various antibacterial agents and antibiotics, and characteristics of the patients and isolates from them each year. Results obtained from these investigations are discussed. In 23 institutions around the entire Japan, 567 strains of presumably etiological bacteria were isolated mainly from the sputa of 459 patients with lower respiratory tract infections during the period from October 1995 to September 1996. MICs of various antibacterial agents and antibiotics were determined against 74 strains of Staphylococcus aureus, 82 strains of Streptococcus pneumoniae, 104 strains of Haemophilus influenzae, 85 strains of Pseudomonas aeruginosa (non-mucoid strains), 18 strains of Pseudomonas aeruginosa (mucoid strains), 52 strains of Moraxella subgenus Branhamella catarrhalis, 25 strains of Klebsiella pneumoniae etc., and the drug susceptibilities of these strains were assessed except for those strains that died during transportation. 1) S. aureus. S. aureus strains for which MICs of oxacillin (MPIPC) were higher than 4 micrograms/ml (methicillin-resistant S. aureus) accounted for 52.7%. Arbekacin (ABK) showed the most highest activity against S. aureus with MIC80 of 0.5 micrograms/ml. Vancomycin (VCM) showed the next highest activity with MIC80 of 1 microgram/ml. These drugs showed the high activities against MRSA with MIC80S of 1 microgram/ml. 2) S. pneumoniae. Most of drugs tested showed potent activities against S. pneumoniae. Imipenem (IPM) and panipenem (PAPM), carbapenems, showed the most potent activity with MIC80S of 0.063 microgram/ml. Cefotaxime (CTX), cefmenoxime (CMX) and cefpirome (CPR) of cephems showed the next most potent activities with MIC80S of 0.25 microgram/ml. Erythromycin (EM) and clindamycin (CLDM) showed low activities with MIC80S 128 micrograms/ml or high. Among these strains, however, 48.8% and 65.9% of respective strains were quite toward sensitive these agents with MICs of 0.063 microgram/ml. 3) H. influenzae. The activities of all drugs were potent against H. influenzae test with all MICs at 4 micrograms/ml or below. Cefotiam (CTM), CMX, cefditoren (CDTR) and ofloxacin (OFLX) showed the most potent activity with MIC90S to 0.063 microgram/ml. 4) P. aeruginosa. (mucoid strains) IPM and tobramycin (TOB) showed the most potent activity against P. aeruginosa (mucoid strains) with MIC80S of 1 microgram/ml. Ceftazidime (CAZ), cefsulodin (CFS) and carumonam (CRMN) showed next potent activity, with MIC80S of 2 micrograms/ml. The MIC80S of the other drugs ranged from 4 micrograms/ml to 32 micrograms/ml. 5) P. aeruginosa (non-mucoid strains). TOB and ciprofloxacin (CPFX) showed the most potent activities against P. aeruginosa (non-mucoid strains) with MIC80S of 1 microgram/ml. The MIC80 of ampicillin (ABPC) was 128 micrograms/ml in 1994, it was 16 micrograms/ml in 1995. 6) K. pneumoniae. All drugs except ABPC were active against K. pneumoniae. CPR and CRMN showed the most potent activities against K. pneumoniae with MIC80S of 0.063 microgram/ml. The MIC80S of the other drugs ranged from 0.125 microgram/ml to 2 micrograms/ml. 7) M. (B.) catarrhalis. Against M. (B.) catarrhalis, all the drugs showed good activities with MIC80S at 4 micrograms/ml or below. And MICs of all strains were 8 micrograms/ml or below. IPM, OFLX and minocycline (MINO) showed the most potent activity with MIC80S of 0.063 microgram/ml. Also, we investigated year to year changes in the characteristics of patients, their respiratory infectious diseases, and the etiology. Patients' backgrounds were examine for 567 isolates from 459 cases. The examination of age distribution found that the proportion of patients with ages over 60 years was 66.3% of all the patients showing a slight increase over that in 1994. Proportion of differe

[Effects of the Prevotella intermedia culture filtrate and short-chain fatty acids on human polymorphonuclear neutrophil functions].

Anaerobes, in particular Prevotella spp., are the predominant bacteria present in mixed respiratory infections. To study the virulence factor of Prevotella intermedia, we examined the effects of the culture filtrate of P. intermedia on the bactericidal activity of human polymorphonuclear neutrophils (PMNs). The culture filtrate significantly inhibited the phagocytic killing and the intracellular killing of Staphylococcus aureus by PMNs (p < 0.05), but the phagocytosis of PMNs was not affected. The boiled culture filtrate also inhibited the phagocytic killing of PMNs (p < 0.05), which suggests that the inhibitory effect may be mediated by heat-stable substances. Succinic and isovaleric acid were mainly detected from the culture filtrate by gas chromatography. The migration of PMNs was significantly inhibited by these short-chain fatty acids (15 mM succinic acid, p < 0.01 and 1 mM isovaleric acid, p < 0.05). The phagocytic killing of PMNs was significantly inhibited by succinic acid (30 mM, p < 0.05) and not by isovaleric acid. Therefore, these inhibitory effects on human PMN functions may be contributed to the virulence of the mixed infections with anaerobes.

Prinzmetal's variant form of angina as a manifestation of alpha-adrenergic receptor-mediated coronary artery spasm: documentation by coronary arteriography.

In four patients with Prinzmetal's variant form of angina, the attack was induced by the combined administration of epinephrine (0.4 to 0.5 mg, given subcutaneously at 7:30 to 8:00 A.M.) and propranolol (40 mg. given orally at 5:00 A.M.). Selective coronary cinearteriography was done before, during, and after the attack with constant monitoring of the ECG and blood pressure. Severe spasm of the right coronary artery occurred at the proximal portion in association with ST-segment elevation in Lead III during the attack and disappeared with the subsidence of the attack in all of them. These results strongly suggest that severe spasm of a large coronary artery mediated by alpha-adrenergic receptors is responsible for the attack of Prinzmetal's variant form of angina.