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Various Time-points (various + time-point)
Selected AbstractsRetinoid- and carotenoid-enriched diets influence the ontogenesis of the immune system in miceIMMUNOLOGY, Issue 2 2003Ada L. Garcia Summary Vitamin A (VA) has been identified as an important factor for the development of the immune system, especially during ontogenesis. It has been shown that antibody secretion and proliferation of lymphocyte populations depend on retinoids. In the present study we investigated the influence of a base VA diet and diets enriched with VA, ,-carotene and lycopene, on the ontogenesis of the immune system in mice. We examined the absolute and relative concentrations of splenic B lymphocytes (CD45R/B220), T lymphocytes (CD3+) and their subpopulations (CD4+ and CD8+), and measured serum immunoglobulin G (IgG) concentrations in the offspring of supplemented dams at different ages (1, 3, 5, 7, 14, 21 and 65 days). The experimental diets resulted in higher numbers of T and B lymphocytes after VA and carotenoid enrichment, when compared, at various time-points, with the base diet. Higher values of total serum IgG were found in the ,-carotene-enriched diet group on day 7. On days 7 and 14, the enriched diets induced significant alterations in the percentages and total numbers of splenic lymphocytes in comparison to the base diet. Our results confirm that supplementation with VA and carotenoids affect the immune-cell function during ontogenesis and suggest a possible role of these nutritional factors on the development of the immune system. [source] Selecting for development of fluoroquinolone resistance in a Campylobacter jejuni strain 81116 in chickens using various enrofloxacin treatment protocolsJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2010K. Stapleton Abstract Aims:, To determine the effect of various enrofloxacin dose regimes on the colonization and selection of resistance in Campylobacter jejuni strain 81116P in experimentally colonized chickens. Methods and Results:, Two experiments were undertaken, in which 14-day-old chickens were colonized with 1 × 107,1 × 109 CFU g,1Camp. jejuni strain 81116P and then treated with enrofloxacin at 12,500 ppm in drinking water for various times. Caecal colonization levels were determined at various time-points after start-of-treatment, and the susceptibility of recovered isolates to ciprofloxacin was monitored. Resistance was indicated by growth on agar containing 4 ,g ml,1 ciprofloxacin, MICs of 16 ,g ml,1 and the Thr86Ile mutation in gyrA. Enrofloxacin at doses of 12,250 ppm reduced Camp. jejuni colonization over the first 48,72 h after start-of-treatment. The degree of reduction in colonization was dose, but not treatment time, dependent. In all cases, maximal colonization was re-established within 4,6 days. Fluoroquinolone-resistant organisms were recoverable within 48 h of start-of-treatment; after a further 24 h all recovered isolates were resistant. In contrast, a dose of 500 ppm enrofloxacin reduced colonization to undetectable levels within 48 h, and the treated birds remained Campylobacter negative throughout the remaining experimental period. By high pressure liquid chromatography, for all doses, the maximum concentrations of enrofloxacin and ciprofloxacin in the caecal contents were detected at the point of treatment completion. Thereafter, levels declined to undetectable by 7 days post-treatment withdrawal. Conclusions:, In a model using chickens maximally colonized with Camp. jejuni 81116P, treatment with enrofloxacin, at doses of 12,250 ppm in drinking water, enables the selection, and clonal expansion, of fluoroquinolone-resistant organisms. However, this is preventable by treatment with 500 ppm of enrofloxacin. Significance and impact of the study:, Treatment of chickens with enrofloxacin selects for resistance in Camp. jejuni in highly pre-colonized birds. However, a dose of 500 ppm enrofloxacin prevented the selection of resistant campylobacters. [source] Relationship of Porphyromonas gingivalis with glycemic level in patients with type 2 diabetes following periodontal treatmentMOLECULAR ORAL MICROBIOLOGY, Issue 4 2008N. Makiura Introduction:, The aim of this study was to assess the relationship between serum glycemic levels and subgingival microbial profile alteration following periodontal treatment in patients with type 2 diabetes mellitus. Methods:, We studied 30 periodontitis patients with type 2 diabetes mellitus who received full-mouth subgingival debridement by analyzing their subgingival microbial profiles using a polymerase chain reaction method at baseline and various time-points for 12 months following treatment. Concurrently, probing pocket depth, bleeding on probing, and metabolic parameters, including glycated hemoglobin A1c (HbA1c), blood sugar level, C-reactive proteins, total cholesterol, triglyceride, and high-density and low-density lipoprotein cholesterol, were recorded. Results:, Periodontal conditions were significantly improved after treatment, and the occurrence rates of periodontal bacterial species, including Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, and Prevotella intermedia, were also reduced. Interestingly, P. gingivalis was detected more frequently in subjects with increased HbA1c values after periodontal treatment than in those patients with decreased HbA1c values. Furthermore, P. gingivalis with type II fimbriae was detected only in HbA1c-increased subjects, while improvements in HbA1c values were observed only in subjects without type II clones. Conclusions:, These results suggest that glycemic level in diabetes is affected by the persistence of P. gingivalis, especially clones with type II fimbriae, in periodontal pockets. [source] A high throughput drug screen based on fluorescence resonance energy transfer (FRET) for anticancer activity of compounds from herbal medicineBRITISH JOURNAL OF PHARMACOLOGY, Issue 3 2007H Tian Background and purpose: We report the development of a very efficient cell-based high throughput screening (HTS) method, which utilizes a novel bio-sensor that selectively detects apoptosis based on the fluorescence resonance energy transfer (FRET) technique. Experimental approach: We generated a stable HeLa cell line expressing a FRET-based bio-sensor protein. When cells undergo apoptosis, they activate a protease called ,caspase-3'. Activation of this enzyme will cleave our sensor protein and cause its fluorescence emission to shift from a wavelength of 535 nm (green) to 486 nm (blue). A decrease in the green/blue emission ratio thus gives a direct indication of apoptosis. The sensor cells are grown in 96-well plates. After addition of different chemical compounds to each well, a fluorescence profile can be measured at various time-points using a fluorescent plate reader. Compounds that can trigger apoptosis are potential candidates as anti-cancer drugs. Key results: This novel cell-based HTS method is highly effective in identifying anti-cancer compounds. It was very sensitive in detecting apoptosis induced by various known anti-cancer drugs. Further, this system detects apoptosis, but not necrosis, and is thus more useful than the conventional cell viability assays, such as those using MTT. Finally, we used this system to screen compounds, isolated from two plants used in Chinese medicine, and identified several effective compounds for inducing apoptosis. Conclusions and Implications: This FRET-based HTS method is a powerful tool for identifying anti-cancer compounds and can serve as a highly efficient platform for drug discovery. British Journal of Pharmacology (2007) 150, 321,334. doi:10.1038/sj.bjp.0706988 [source] Evaluation of vitreous levels of gatifloxacin after systemic administration in inflamed and non-inflamed eyesACTA OPHTHALMOLOGICA, Issue 6 2009Rajpal Abstract. Purpose:, This study aimed to evaluate the human vitreous penetration of gatifloxacin in inflamed and non-inflamed eyes after oral administration. Methods:, Vitreous penetration of single-dose (400 mg) oral gatifloxacin was evaluated in patients (n = 33) undergoing vitreous tap during the standard procedure for intravitreal antibiotic injection for acute postoperative endophthalmitis at various time-points. Vitreous penetration of 400 mg oral gatifloxacin was evaluated in the non-inflamed eyes of patients (n = 33) undergoing pars plana vitrectomy at similar time-points. The study was extended to evaluate the vitreous penetration of single-dose oral (800 mg) gatifloxacin at a single time-point in inflamed (n = 10) and non-inflamed (n = 11) eyes. Results:, After 400 mg oral gatifloxacin, inflamed eyes showed mean vitreous concentrations of 0.58±0.19,g/ml, 1.33±0.33 ,g/ml and 1.30 ± 0.23 ,g/ml at 2, 4 and 6 hours, respectively. The levels reached at 2 and 4 hours were found to be significantly increased compared with those in non-inflamed eyes. At the 800-mg dose, 4-hour vitreous levels in inflamed and non-inflamed eyes were 1.57 ± 0.3 ,g/ml and 1.42 ± 0.24 ,g/ml, respectively. Although the increased dose of gatifloxacin elevated plasma concentration, it failed to raise vitreous levels significantly higher than the 400-mg dose at the 4-hour time-point. Conclusions:, Orally administered gatifloxacin achieves therapeutic levels in both inflamed and non-inflamed human eyes with a spectrum covering the bacterial species most frequently involved in the various causes of endophthalmitis. However, the levels achieved were below the MIC90 for Pseudomonas aureginosa and Enterococcus. [source] In vivo imaging of microglial cell traffickingACTA OPHTHALMOLOGICA, Issue 2009M PAQUES Purpose Microglial cells (MCs) are active sensors of neural tissues that are rapidly mobilized upon disruption of homeostasis. OUr goal was to observe in vivo the migration of MCs, which has not been done yet. Methods Following acute laser damage, the behavior of MCs in the retina of adult Cx3cr1gfp/+ and gfp/gfp mice was observed noninvasively using time-lapse confocal scanning laser ophthalmoscopy. Observation were done at various time-points up to 8 days after laser damage. Results Focal damage elicite prompt migratory response of MCs within 200 to 400 µm around laser burns. This migratory response was preceded in all case by dendritic reorientation. Convergent and nonconvergent migration were observed. Such migratory activity persisted several days after laser damage. At day 8, the microglia network was restored and microglial locomotion had ceased. Conclusion To our knowledge, this is the first observation of microglial locomotion in vivo. A Morphological evidence of microglial activation starts with dendritic reorganization. Migrating cells were only of the dendritic type (i.e. not ameboid). There appears to be a notable heterogeneity in the locomotor response of MCs. MCs within and around scars remain highly motile and mobile several days after laser damage. [source] |