Enhanced vaccination effect against influenza by prebiotics in elderly patients receiving enteral nutrition.

AIM: We investigated the effect of prebiotics on the immunological response after influenza vaccination in enterally fed elderly individuals. The intervention group was given an enteral formula containing lactic acid bacteria-fermented milk products. In addition, two different types of other prebiotics, galacto-oligosaccharide and bifidogenic growth stimulator, were also given. The two prebiotics improved intestinal microbiota differently. In a control group, a standard formula without prebiotics was given. METHODS: An enteral formula with (intervention group [F]) or without (control group [C]) prebiotics was given through percutaneous endoscopic gastrostomy to elderly participants for 10 weeks. Influenza vaccine was inoculated at week 4. Nutritional and biochemical indices, intestinal micro bacteria and immunological indices were analyzed. RESULTS: The Bifidobacterium count in groups F and C at week 0 was 6.4 ± 1.9 and 6.6 ± 3.0 (log10 [count/g feces]), respectively. Although the count in group C decreased at week 10, the count in group F increased. The Bacteroides count in group F increased from 10.7 ± 0.9 to 11.4 ± 0.5, but decreased in group C from 11.2 ± 0.2 to 10.7 ± 0.4. Although the enhanced titers of H1N1, H3N2 and B antigens against the vaccine decreased thereafter in group C, these enhanced titers in group F could be maintained. CONCLUSION: Our findings suggest that prebiotics affect the intestinal microbiota and might maintain the antibody titers in elderly individuals.

Clinical effects of probiotic Bifidobacterium longum BB536 on immune function and intestinal microbiota in elderly patients receiving enteral tube feeding.

BACKGROUND: Immune system function declines with age. We evaluated the effects of supplementation with the probiotic Bifidobacterium longum BB536 on immune function and intestinal microbiota in the elderly. MATERIALS AND METHODS: In a double-blind study, 45 elderly patients fed by enteral tube feeding (mean [SD] age 81.7 [8.7] years) were given BB536 (n = 23) or a placebo powder (n = 22) for 12 weeks and were observed for an additional 4 weeks posttreatment. At week 4, all patients received an influenza vaccination (A/H1N1, A/H3N2, and B). Clinical data were assessed, including body temperature, bowel movements, fecal microbiota, and immunological biomarkers in blood. RESULTS: BB536 intake significantly increased cell numbers of bifidobacteria in fecal microbiota. There was a tendency toward an increase (P = .085 at week 4 and P = .070 at week 16) of serum IgA in the BB536 group compared with the placebo group. BB536 intake did not significantly affect hemagglutination inhibition (HI) titers in response to the influenza vaccine. Natural killer (NK) cell activity decreased significantly in the placebo group during the intervention but not in the BB536 group. Among those subjects with low NK cell activity (<55%, n = 10 for each group), a significant intergroup difference (P < .05) was observed in the changed values from baseline of NK cell activity at weeks 8 and 12. CONCLUSIONS: These results shed new light on the potential of long-term ingestion of BB536 in increasing the cell number of bifidobacteria in intestinal microbiota and modulating immune function in the elderly.

Effects of peripheral cannabinoid receptor ligands on motility and polarization in neutrophil-like HL60 cells and human neutrophils.

The possible role of the peripheral cannabinoid receptor (CB2) in neutrophil migration was investigated by using human promyelocytic HL60 cells differentiated into neutrophil-like cells and human neutrophils isolated from whole blood. Cell surface expression of CB2 on HL60 cells, on neutrophil-like HL60 cells, and on human neutrophils was confirmed by flow cytometry. Upon stimulation with either of the CB2 ligands JWH015 and 2-arachidonoylglycerol (2-AG), neutrophil-like HL60 cells rapidly extended and retracted one or more pseudopods containing F-actin in different directions instead of developing front/rear polarity typically exhibited by migrating leukocytes. Activity of the Rho-GTPase RhoA decreased in response to CB2 stimulation, whereas Rac1, Rac2, and Cdc42 activity increased. Moreover, treatment of cells with RhoA-dependent protein kinase (p160-ROCK) inhibitor Y27632 yielded cytoskeletal organization similar to that of CB2-stimulated cells. In human neutrophils, neither JWH015 nor 2-AG induced motility or morphologic alterations. However, pretreatment of neutrophils with these ligands disrupted N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)-induced front/rear polarization and migration and also substantially suppressed fMLP-induced RhoA activity. These results suggest that CB2 might play a role in regulating excessive inflammatory response by controlling RhoA activation, thereby suppressing neutrophil migration.

Problems in blood alcohol testing of severely injured drivers brought to emergency departments in Japan.

Breath alcohol tests are widely used to control DUI (driving under the influence) in Japan. However, this test is not applied to injured drivers transported to emergency hospitals. In such cases, BAC (blood alcohol concentration) testing should be done to prove DUI. In this paper, we tried to clarify two important issues on the BAC testing in Japan using a questionnaire survey and experiments about contamination of antiseptic ethanol. First, we have described the doctor's dilemma with DUI cases; our present questionnaire survey showed that the police often request the doctor to volunteer blood samples of the suspected drunk drivers brought to emergency hospitals since they have not been granted the right to order blood sampling in Japan. Then, doctors face a serious dilemma whether comply with the police request or not, resulting in widely different responses. Secondly, we have estimated the effects of antiseptic ethanol routinely used as a dermal antiseptic on the BAC tests. Our present experiments showed that uptake of ethanol can occur under certain conditions. Given the actual conditions outlined in the questionnaire, there seem to be a definite risk of ethanol contamination in BAC testing. Obviously, the time has come to discuss problems in BAC testing of injured drivers brought to emergency hospitals in Japan.

Protein-tyrosine kinase, Syk, is required for CXCL12-induced polarization of B cells.

Cell polarization and migration in response to CXCL12 is essential for hematopoiesis. To investigate the role of Syk in CXCL12/CXCR4-induced signaling, wild-type Syk or its dominant-negative form (DN-Syk) was introduced in mouse pro-B cells, BAF3. With CXCL12 stimulation, BAF3 cells became polarized with the formation of a leading edge and contractile uropod at the rear end with increased motility. Overexpression of wild-type Syk caused enhanced polarization, whereas DN-Syk inhibited cell polarity due to the loss of contractile structure at the rear end, and the altered phenotype was enhanced after CXCL12 stimulation. Motility of mutant BAF3 containing DN-Syk increased independent of CXCL12 stimulation. As beta1 integrin-mediated cell adhesion was inhibited, decreased adhesion might promote motility. CXCL12 stimulation led to prompt activation of RhoA, but expression of DN-Syk suppressed RhoA activation. These results demonstrate that Syk participates in CXCL12-induced cell polarization, which occurs in concert with cell adhesion mediated by beta1 integrin.

Mutations in 14 Y-STR loci among Japanese father-son haplotypes.

In the present study 161 Japanese father/son haplotype transfers in 147 pedigrees were analyzed at 14 Y-STRs with two multiplex PCR-based typing systems. Five isolated single repeat mutations were identified at the DYS389I, DYS439, Y-GATA-H4, DYS389II and DYS391 loci, and a pedigree showing triple alleles at the DYS385 locus (a duplicate locus) without allelic discrepancy between the father and son was also observed. The overall mutation rate estimated across the 14 Y-STRs in the Japanese population was 0.22%/locus/meiosis (95% C.I. 0.09-0.51%). This rate was not significantly different (p>0.05) from those of autosomal STRs and Y-STRs in other populations, including German, Austrian, Polish and Norwegian populations. Furthermore, 138 haplotypes were identified in 147 pedigrees with a haplotype diversity value of 0.9983. Therefore, a combination of the two systems should permit effective analysis with sufficient discriminatory power.

Sensitive determination of pethidine in body fluids by gas chromatography-tandem mass spectrometry.

We have presented a simple and sensitive method for determining pethidine, a narcotic analgesic drug in body fluids by gas chromatography-tandem mass spectrometry (GC-MS/MS). Pethidine and 4'-piperidinoacetophenone (internal standard) were extracted from body fluids with Bond Elut C(18) columns; the recoveries were above 85% for both compounds. The calibration curves for blood and urine showed good linearities in the range of 1.25-40 ng/ml. Its detection limits (signal-to-noise ratio=3) were estimated to be approximately 0.5 ng/ml of whole blood and urine.

Haplotype analysis with 14 Y-STR loci using 2 multiplex amplification and typing systems in 2 regional populations in Japan.

In this study 14 Y-STR loci (DYS393, DYS19, DYS391, DYS437, DYS435, DYS439, DYS389II, DYS438, DYS436, DYS390, Y-GATA-H4, DYS385, Y-GATA-A7.1 and DYS392) were analysed in 207 Japanese males from Honshu (main island of Japan, Nagoya City) and 87 Japanese males from Okinawa (southernmost islands of Japan) using two multiplex PCR typing systems, a novel 10-plex amplification system and a new commercially available 6-plex typing kit which had two loci in common. The allele frequency distributions were similar at almost all of the 14 loci. Of the haplotypes observed, 244 were unique in both Japanese populations and 17 haplotypes were observed more than once but the 2 populations shared only 7 haplotypes. The haplotype diversities for the 14 loci were 0.9987 and 0.9976 in Honshu and Okinawa Japanese, respectively. The haplotype analysis at 14 Y-STR loci would be useful for personal identification in forensic fields and for population genetics because of the high divergence of these haplotypes.

Identification and characterization of 17 phenothiazine compounds by capillary high-performance liquid chromatography/fast atom bombardment mass spectrometry.

Phenothiazines are widely prescribed as neuroleptics; some are used as antihistaminics. These compounds are important in clinical and forensic toxicology. Seventeen phenothiazine compounds with heavy side chain structures have been found to be detectable by high-performance liquid chromatography/fast atom bombardment-mass spectrometry (HPLC/FAB-MS) method. Authentic samples of the compounds were subjected to our HPLC/FAB-MS system; their mass spectra were obtained by positive and negative modes. Four typical phenothiazines, in the serum samples of two patients, were also analyzed. All 17 phenothiazines were sufficiently separated on the chromatogram. In the positive mode, all the base peaks were quasimolecular ions; their main fragment ions observed were [M-R(1)+CH(2)](+), [R(1)](+), [M-R(1)](+) and [M+H+Gly](+). In the negative mode, the base peaks were [Cl](-) for chlorpromazine, prochlorperazine and perphenazine, three compounds containing chloride. For the other compounds, they were [M-R(1)-CH(3)](-), [M-R(1)-CH(2)CH(3)](-) or [M-R(1)-(CH(3))(2)](-) ions. We observed [M+H](-) ions in all the compounds, however, the ir intensities were variable (3-74%). The spectra and mass chromatograms of four compounds and their metabolites in the extracted serum samples from two patients, were also obtained. The approximate detection limits for phenothiazines were less than 1 ng on-column in the positive mode, and about 1 microg on-column in the negative mode. We have succeeded in the identification and characterization of 17 phenothiazine compounds at therapeutic concentrations in body fluids using our HPLC/FAB-MS system. The present method would be useful in forensic toxicological practice.