Soft-diet feeding inhibits adult neurogenesis in hippocampus of mice.

Our previous study showed that mice fed on a soft diet after weaning had reduced levels of brain-derived neurotrophic factor (BDNF) protein in the hippocampus after 3 months of age compared with mice fed on a hard diet. BDNF is one of the most effective promoters of neurogenesis in the hippocampus, and enhancement of BDNF production has been shown to enhance neural precursor cell proliferation in the dentate gyrus. We hypothesized that soft-diet feeding during development would reduce the proliferation rate of precursor cells, resulting in lower production of new neurons in the hippocampus. Male C57BL/6 mice pups were fed either a solid (hard-diet group) or powdered (soft-diet group) diet starting at weaning. Three and six months after birth, mice of each group received intraperitoneal injections of bromodeoxyuridine (BrdU, 50 mg/kg body weight), twice a day for 3 consecutive days. After survival time of 1 day, 1 week, or 4 weeks, the mice were anesthetized and perfused transcardially. Newborn cells in the dentate gyrus were examined by immunohistochemistry using anti-BrdU antibody. In addition, phenotypically neuronal cells among the newborn cells were detected by immunofluorescent double labeling for BrdU and mature neuron-specific nuclear protein (NeuN) using anti-BrdU and anti-NeuN antibodies. Total number of BrdU-positive cells in the dentate gyrus was fewer in the 6-month-old mice than in the 3-month-old mice at any survival time investigated, and fewer in the soft-diet group than in the hard-diet group at 3 and 6 months of age. Neither soft-diet feeding nor aging affected ratio of phenotypically neuronal cells among newborn cells. These results indicate that insufficient mastication activity during development as well as aging restrains hippocampal neurogenesis in adulthood.

Effects of soft-diet feeding on BDNF expression in hippocampus of mice.

Our previous study showed that mice fed a soft diet after weaning had reduced synaptic connections in the hippocampal formation and impaired spatial learning ability after 3 months of age. We hypothesized that soft-diet feeding during development reduced levels of brain-derived neurotrophic factor (BDNF) protein in the hippocampus, resulting in lower synaptic densities in this region. Male pups of C57BL/6 mice were fed either a solid (hard-diet group) or powdered diet (soft-diet group), starting at weaning. Expression of BDNF protein in the hippocampus and cerebral cortex was evaluated quantitatively with enzyme-linked immunosorbent assay (ELISA) at 1, 3 and 6 months of age. Reduction in BDNF protein levels due to soft diet was detected markedly in the hippocampus of 3- and 6-month-old mice. On the other hand, a soft diet showed no significant effect on BDNF content in the cerebral cortex throughout the ages investigated. Immunohistochemistry of hippocampal formation in 3-month-old mice revealed that intensities of BDNF immunoreactivity in the dentate gyrus granule cell layer and CA1 and CA3 pyramidal cell layers appeared diminished in mice fed the soft diet compared with mice fed the hard diet. These results indicate that insufficient mastication activity during development reduces BDNF protein levels in the hippocampus and influences synaptic plasticity in this region.

Specific timing of taurine supplementation affects learning ability in mice.

The effects of taurine supplementation on visual discrimination in mice were examined. Taurine, 2-aminoethane-sulphonic acid, found in high concentrations in the central nervous system of mammals and in human milk, has been shown to be essential for development. Male mice were divided into four groups according to taurine supplementation periods. 1) Lifelong: taurine (400 mg/kg/day) was dissolved in distilled water and provided as drinking water. In the prenatal period, taurine was given via the mother. After weaning mice were administered taurine in drinking water. 2) Pre-weaning: mice were exposed to taurine prior to weaning, 3) Post-weaning: mice were exposed to taurine after weaning. 4) CONTROL: no supplementation of taurine. It was shown that the Lifelong group required a longer period of time to acquire visual discrimination than the CONTROL group. Conversely, in the Post-weaning group, mice learned the task faster than CONTROLs. Visual discrimination learning time in the Pre-weaning group showed no significant difference compared with that in the CONTROL group. From these results, we suggest that the perinatal to early postnatal period is a "sensitive period" where taurine supplementation can result in retardation of learning in later life. At the same time, taurine supplementation after weaning improved visual discrimination learning. Thus, timing of taurine supplementation affected learning.

Effectiveness of probiotic therapy for the prevention of relapse in patients with inactive ulcerative colitis.

AIM: To evaluate the effectiveness of probiotic therapy for suppressing relapse in patients with inactive ulcerative colitis (UC). METHODS: Bio-Three tablets, each containing 2 mg of lactomin (Streptococcus faecalis T-110), 10 mg of Clostridium butyricum TO-A, and 10 mg of Bacillus mesentericus TO-A, were used as probiotic therapy. Sixty outpatients with UC in remission were randomly assigned to receive 9 Bio-Three tablets/day (Bio-Three group) or 9 placebo tablets/day (placebo group) for 12 mo in addition to their ongoing medications. Clinical symptoms were evaluated monthly or on the exacerbation of symptoms or need for additional medication. Fecal samples were collected to analyze bacterial DNA at baseline and 3-mo intervals. Terminal restriction fragment length polymorphism and cluster analyses were done to examine bacterial components of the fecal microflora. RESULTS: Forty-six patients, 23 in each group, completed the study, and 14 were excluded. The relapse rates in the Bio-Three and placebo groups were respectively 0.0% vs 17.4% at 3 mo (P = 0.036), 8.7% vs 26.1% at 6 mo (P = 0.119), and 21.7% vs 34.8% (P = 0.326) at 9 mo. At 12 mo, the remission rate was 69.5% in the Bio-Three group and 56.6% in the placebo group (P = 0.248). On cluster analysis of fecal flora, 7 patients belonged to cluster I, 32 to cluster II, and 7 to cluster III. CONCLUSION: Probiotics may be effective for maintaining clinical remission in patients with quiescent UC, especially those who belong to cluster I on fecal bacterial analysis.

Incidence of dehydration encephalopathy among patients with disturbed consciousness at a hospital emergency unit.

Systemic dehydration and diffuse central nervous system signs without any other illness is referred to as dehydration encephalopathy (DE). However, the incidence of DE at emergency units remains uncertain. We investigated the incidence of DE among persons with disturbed consciousness who visited the emergency unit. We reviewed the medical case records of the emergency unit at our university hospital during a 6-month period. Among them, 132 patients presented with disturbed consciousness as the sole initial manifestation on arrival. They were 75 men, 47 women; mean age 68 years (16-95 years). After carefully excluding other etiologies, the incidence of DE was 2% among all persons in the emergency unit and 4% among persons older than 68 years. In conclusion, the incidence of DE in our emergency unit was not common. Nevertheless, recognition of DE is extremely important in order to avoid unnecessary medication in elderly subjects.

Monitoring functional serum antitumor necrosis factor antibody level in Crohn's disease patients who maintained and those who lost response to anti-TNF.

BACKGROUND: Infliximab (IFX) is an antitumor necrosis factor (TNF)-α antibody used to treat Crohn's disease (CD). However, antibodies to IFX (ATI) emerge, which can impair its efficacy. A fluid-phase enzyme immunoassay (FP-EIA) was established for measuring serum functional IFX (f-IFX) in CD patients receiving maintenance IFX. METHODS: In 31 patients, 16 had maintained response (GI) and 15 had lost response to IFX despite good initial response (GII) were selected. Serum f-IFX was measured just before and immediately after IFX infusion and the values together with CD activity index (CDAI) and C-reactive protein (CRP) were compared. RESULTS: IFX therapy in GI and GII were 1.8 ± 1.2 years and 2.7 ± 1.5 years, respectively, while the median dose frequency was 56 days in GI and 29 days in GII. Our FP-EIA for f-IFX showed TNF-α binding increasing with the IFX dose, which was suppressed by antibodies to IFX. On the infusion day, CRP and CDAI in GII were significantly higher than in GI, while median trough f-IFX for GI and GII were 4.7 µg/mL and 6.3 µg/mL, respectively. The median f-IFX immediately after IFX infusion for GI and GII were 149.5 µg/mL and 126.3 µg/mL, respectively (P = 0.0488), and binary logistic regression showed conditional maximum likelihood estimate to be -0.0258 (P = 0.0395), supporting association of low postinfusion f-IFX to the loss of response. CONCLUSIONS: FP-EIA could accurately measure f-IFX. High serum ATI strongly impacted f-IFX levels immediately after an infusion. The postinfusion f-IFX level was associated with clinical response. f-IFX level should be valuable in decision-making to optimize treatment efficacy.

Adacolumn selective leukocyte adsorption apheresis in patients with active ulcerative colitis: clinical efficacy, effects on plasma IL-8, and expression of Toll-like receptor 2 on granulocytes.

Adacolumn selective granulocyte and monocyte apheresis (GMA) depletes activated leukocytes in patients with ulcerative colitis (UC). However, this per se cannot fully explain the efficacy of GMA. We have investigated the effects of GMA on the expression of toll-like receptors (TLRs) and plasma interleukin-8 (IL-8). Twenty-two patients with clinical activity index (CAI) of 5-17, 15 with total colitis and 7 with left-sided colitis, were included. Each patient could receive up to 10 GMA sessions, at 1 or 2 sessions per week. GMA was added to the patients' ongoing medication following a relapse or worsening UC, but no additional medication was given. Further, at entry and pre-GMA, blood samples were taken for full blood cell count, expression of TLRs on leukocytes, and plasma IL-8. Seventy-five percent of patients achieved remission after the 10th session (CAI, < or =4; P < 0.005) and there was a marked fall in C-reactive protein (P < 0.01), plasma IL-8 (P < 0.001), and granulocytes (P < 0.05) but an increase in lymphocytes (P < 0.05). The expression of TLR2 on granulocytes was down-modulated (P < 0.05) together with suppression of inflammatory cytokines produced by peripheral blood leukocytes. In conclusion, GMA appears to be an effective adjunct therapy to induce remission in the majority of patients, who are then spared from excess drug therapy. The procedure is associated with sustained immunomodulation. Control studies should strengthen these findings.

Clinical effectiveness of probiotics therapy (BIO-THREE) in patients with ulcerative colitis refractory to conventional therapy.

OBJECTIVE: Intestinal microflora has been implicated in the etiology of ulcerative colitis (UC). Over the past few years, the use of probiotics in UC has gained attention. The aim of this study was to evaluate the efficacy of probiotics therapy for mild to moderate distal UC refractory to conventional therapies. MATERIAL AND METHODS: Twenty patients with mild to moderate distal UC took 9 BIO-THREE tablets per day for 4 weeks. Clinical symptoms and endoscopic findings were evaluated as ulcerative colitis disease activity index (UCDAI) scores before and after administration of BIO-THREE. Fecal samples were collected from all patients before and after probiotics administration, and fecal microflora was analyzed by the terminal restriction fragment length polymorphism (T-RFLP) method. RESULTS: Remission (UCDAI score < or =2) was observed in 45% (9/20) of the patients; response (decrease in UCDAI > or = 3, but final score > or = 3) in 10% (2/20); no response in 40% (8/20); and worsening (UCDAI > 3) in 5% (1/20). T-RFLP analysis indicated that the principal alteration in microflora was an increase in bifidobacteria. CONCLUSIONS: This study showed that administration of BIO-THREE improved the clinical symptoms and endoscopic findings in patients with UC, indicating that administration of BIO-THREE is safe and efficacious for the treatment of UC.