Effects of naringin on hydrogen peroxide-induced cytotoxicity and apoptosis in P388 cells

Flavonoids are widely recognized as naturally occurring antioxidants. Naringin (NG) is one of the flavonoid components in citrus fruits such as grapefruit. Hydrogen peroxide (H2O2) causes cytotoxicity through oxidative stress and apoptosis. In this paper, we examined the effects of NG on H2O2-induced cytotoxicity and apoptosis in mouse leukemia P388 cells. Cytotoxicity was determined by mitochondrial activity (MTT assay). Apoptosis and DNA damage were analyzed by measuring chromatin condensation and Comet assay (alkaline single cell gel electrophoresis), respectively. H2O2-induced cytotoxicity was significantly attenuated by NG or the reduced form of glutathione (GSH), a typical intracellular antioxidant. NG suppressed chromatin condensation and DNA damage induced by H2O2. These results indicate that NG from natural products is a useful drug having antioxidant and anti-apoptopic properties.     

Eight novel single nucleotide polymorphisms in ABCG2/BCRP in Japanese cancer patients administered irinotacan

Eight novel single nucleotide polymorphisms (SNPs) were found in the gene encoding the ATP-binding cassette transporter, ABCG2/BCRP, from 60 Japanese individuals administered the anti-cancer drug irinotecan. The detected SNPs were as follows: 1) SNP, MPJ6_AG2005 (IVS2-93T>C); Gene Name, ABCG2; Accession Number, NT_006204; 2) SNP, MPJ6_AG2007 (IVS3+71_72 insT); Gene Name, ABCG2; Accession Number, NT_006204; 3) SNP, MPJ6_AG2012 (IVS6-204C>T); Gene Name, ABCG2; Accession Number, NT_006204; 4) SNP, MPJ6_AG2015 (at nucleotide 1098G>A (exon 9) from the A of the translation initiation codon); Gene Name, ABCG2; Accession Number, NT_006204; 5) SNP, MPJ6_AG2017 (1291T>C (exon 11)); Gene Name, ABCG2; Accession Number, NT_006204; 6) SNP, MPJ6_AG2019 (IVS11-135G>A); Gene Name, ABCG2; Accession Number, NT_006204; 7) SNP, MPJ6_AG2020 (1465T>C (exon 12)); Gene Name, ABCG2; Accession Number, NT_006204; 8) SNP, MPJ6_AG2023 (IVS13+65T>G); Gene Name, ABCG2; Accession Number, NT_006204.MPJ6_AG2015 was a synonymous SNP (E366E). MPJ6_AG2017 and MPJ6_AG2020 resulted in amino acid alterations, F431L and F489L, respectively.     

Prevalence of ciliochoroidal effusion after prophylactic laser iridotomy

PURPOSE: To study the prevalence of ciliochoroidal effusion (CE) after prophylactic laser iridotomy (LI). METHODS: An ultrasound biomicroscope (UBM) examination was performed before, 2, and 24 hours after LI to investigate the changes in supraciliary-choroidal space after LI in circumference of 38 eyes in 23 chronic primary angle-closure patients. Subjects were divided randomly into two groups based on LI settings: 21 eyes by argon laser (Ar-LI group) and argon combined with yttrium-aluminum-garnet (YAG) laser in 17 eyes (Ar-YAG-LI group).Ciliochoroidal effusion was observed predominantly in the inferior sector (12 of 12 eyes, 100%) and rare in nasal part of the eyes (4 of 12 eyes, 33%). Ciliochoroidal effusion appeared more often in the Ar-LI group (10 eyes, 52%) than in the Ar-YAG-LI group (2 eyes, 12%). All CE disappeared 7 days after by UBM observation. CONCLUSIONS: Subclinical CE observed by UBM frequently occurs after LI.     

Spontaneous closure of stage 2 macular hole observed with optical coherence tomography

PURPOSE: To report the clinical course and images of optical coherence tomography of an eye with a stage 2 macular hole which closed spontaneously. DESIGN: Observational case report. METHODS: Serial optical coherence tomographic images were obtained. RESULTS: A 67-year-old woman received a diagnosis of stage 2 idiopathic macular hole in her right eye. The diagnosis was made with a tomographic image of a full-thickness dehiscence of the neurosensory retina at the fovea. The posterior hyaloid membrane was adhering to the edge of the dehiscence. Four weeks later, the closure of the macular hole was ascertained with optical coherence tomography. The posterior hyaloid membrane was fully separated from the fovea. CONCLUSION: A stage 2 macular hole may close spontaneously with the separation of the hyaloid membrane.     

Characterization of beta-adrenergic receptor sequestration by newly developed whole cell binding assays

Upon agonist binding, beta-adrenergic receptors sequestrate from the cell surface plasma membrane to cytosol. In the present study, we examine the kinetics of sequestration of beta1-adrenergic receptor and beta3-adrenergic receptor subtypes by radioligand binding assays using whole cells ('whole cell binding assays'). We found that HEK293T cells, but not COS1 cells, were readily and uniformly detached from the culture dish upon exposure to ice-cold phosphate-buffered saline. Using this property of HEK293T cells, we conducted whole cell binding assays using a hydrophilic antagonist ([3H]CGP-12177) and HEK293T cells transiently overexpressing human beta1-adrenergic receptor or beta3-adrenergic receptor. The Bmax and Kd values were 5.96 +/- 0.97 pmol/mg protein and 1 +/- 0.23 nM for the beta1-adrenergic receptor, and were 1.84 +/- 0.13 pmol/mg protein and 44.7 +/- 2.5 nM for the beta3-adrenergic receptor, respectively. Isoproterenol treatment, but not 6-[3-(dimethylamino)propionyl]forskolin treatment, for 2 h resulted in a dose-dependent loss of the number of the cell surface beta1-adrenergic receptor. At 100 microM, 36.6 +/- 5.7% of the cell surface beta1-adrenergic receptor was lost. In contrast, the cell surface beta3-adrenergic receptor number remained unchanged with isoproterenol treatment. Thus, beta1-adrenergic receptor sequestrates upon agonist stimulation but the same agonist stimulation does not induce beta3-adrenergic receptor sequestration, as demonstrated by our whole cell binding assays.     

Measurement of residual 152Eu activity induced by atomic bomb neutrons in Nagasaki and the contribution of environmental neutrons to this activity

Residual 152Eu activities induced by neutrons from the Nagasaki atomic bomb were measured for nine mineral samples located up to 1,061 m in the slant range and one control sample at 2,850 m from the hypocenter. A chemical separation to prepare europium-enriched samples was performed for all samples, and gamma ray measurements were carried out with a low background well-type germanium detector. In this paper, the measured specific activities of 152Eu are compared with activation calculations based on the DS86 neutron fluence and the 93Rev one. The calculated-to-measured ratios are also compared with those of 60Co and 36Cl. The present results indicate that the measurements agree to the calculation within a factor of three as observed in the nuclear tests at Nevada. The activation level of environmental neutrons and the detection limit for 152Eu are also discussed.     

Wild and synanthropic hosts of Leishmania (Viannia) braziliensis in the endemic cutaneous leishmaniasis locality of Amaraji, Pernambuco State, Brazil

Evidence of Leishmania infection was found in small mammals captured between 1996 and 2000 in the Amaraji region, Pernambuco State, Brazil. The kDNA polymerase chain reaction (PCR), using primers specific for subgenus L. (Viannia), was positive for 43/153 water rats (Nectomys squamipes), 13/81 black rats (Rattus rattus), 15/103 grass mice (Bolomys lasiurus), 1/14 marsh mice (Holochilus scieurus), 2/50 field mice (Akodon arviculoides), 2/12 woolly opossums (Marmosa sp.), and 5/37 common opossums (Didelphis albiventris). This same kDNA PCR was positive for 12/61 dog and 8/58 horse skin samples. In paired PCR tests of 203 small mammals, 18.7% were positive with the kDNA primers and 18.2% with rDNA primers. Amastigotes were seen in 26/460 and L. (V.) braziliensis was isolated from 5 grass mice and 1 black rat. We concluded that small mammals, particularly rodents, are infected with parasites of the subgenus L. (Viannia). The isolation of L. (V.) braziliensis zymodeme IOC/Z74 from 6 rodents and the fact that all the other described L. (Viannia) species that commonly infect humans have never been found in rodents or marsupials leads us to suggest that the positive PCRs indicate infections of L. (V.) braziliensis. The isolation of zymodeme IOC/Z74 from humans reinforces our hypothesis that small, ground-loving mammals, such as rodents are the primary reservoirs of L. (V.) braziliensis.     

Randomized controlled trial of the effect of bifidobacteria-fermented milk on ulcerative colitis

BACKGROUND: Alterations of intestinal flora, such as reduction in the concentration of bifidobacteria and increase in that of Bacteroides species, are apparently associated with the severity of ulcerative colitis. OBJECTIVE: We conducted a randomised clinical trial of the use of a bifidobacteria-fermented milk (BFM) supplement as a dietary adjunct in the treatment of ulcerative colitis. METHODS: The subjects were randomly divided into two groups: a group with BFM supplementation (BFM group, 11 subjects) and a control group (control group, 10 subjects). The BFM group was given 100 mL/day of BFM for one year. Colonoscopies, general blood markers and examinations of intestinal flora including the analysis of fecal organic acids were performed at the commencement of the study and after one year. RESULTS: Exacerbation of symptoms was seen in 3 out of 11 subjects in the BFM group and in 9 out of 10 in the control group. Log rank statistic analysis of the cumulative exacerbation rates showed a significant reduction in exacerbations for the BFM group (p = 0.0184). The analysis of microflora and the organic acids in the feces showed a significant reduction in the relative proportion of B. vulgatus in Bacteroidaceae and butyrate concentration, respectively, after supplementation with BFM, in comparison with before. CONCLUSION: Supplementation with the BFM product was successful in maintaining remission and had possible preventive effects on the relapse of ulcerative colitis.     

Changing prevalent T serotypes and emm genotypes of Streptococcus pyogenes isolates from streptococcal toxic shock-like syndrome (TSLS) patients in Japan

We surveyed T serotypes and emm genotypes of Streptococcus pyogenes isolates from streptococcal toxic shock-like syndrome (TSLS) patients. T1 (emm1) remained dominant through 1992 to 2000, but the dominant T3 (emm3.1) strains from 1992 to 1995 disappeared during 1996-2000. Strains of several emm genotypes emerged during 1996-2000, indicating alterations in the prevalent strains causing TSLS.