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We tested a hypothesis that manganese enhanced magnetic resonance imaging (MEMRI) after systemic injection of MnCl2 could detect axonal sprouting in the hippocampus following kainate (KA) induced status epilepticus (SE). MEMRI was performed at 3 h, 25 h, 4 days, and 2 months post-SE. To assess the contribution of various cellular alterations that occur in parallel with sprouting to the MEMRI signal, we sacrificed animals for histology at 4 days and 2 months post-SE. Neurodegeneration was assessed from thionin and Fluoro-Jade B stained preparations, astrogliosis from GFAP (glial fibrillary acidic protein) and microgliosis from Ox-42 immunostained preparations. Sprouting of granule cells axons (mossy fibers) in the dentate gyrus was analyzed from Timm stained sections. Occurrence of spontaneous epileptic seizures was analyzed at 2 months post-SE using continuous video-EEG monitoring. Integrity of the blood–brain barrier (BBB) was studied using Gd-enhanced MRI. We found abnormal MEMRI hyperintensity in the CA1 and the dentate gyrus at 2 months post-SE but not at earlier time points. Based on histologic analysis of individual animals with MEMRI hyperintensity, hippocampal MEMRI changes could be attributed to increasing axonal density rather than to neurodegeneration, astrogliosis, or microgliosis. Moreover, MEMRI contrast was not affected by seizure activity, and we could not detect any leakage of the BBB that could have explained the observed MEMRI hyperintensity. Present data show that systemic MEMRI can reveal axonal sprouting, and thus, can potentially serve as a marker for neuroplasticity in preclinical studies.

In traumatic brain injury (TBI) the initial impact causes both immediate damage and also launches a cascade of slowly progressive secondary damage. The chronic outcome disabilities vary greatly and can occur several years later. The aim of this study was to find predictive factors for the long-term outcome using multiparametric, non-invasive magnetic resonance imaging (MRI) methodology and a clinically relevant rat model of fluid percussion induced TBI. Our results demonstrated that the multiparametric quantitative MRI (T 2, T 1ρ, trace of the diffusion tensor D av, the extent of hyperintense lesion and intracerebral hemorrhage) acquired during acute and sub acute phases 3 h, 3 days, 9 days and 23 days post-injury has potential to predict the functional and histopathological outcome 6 to 12 months later. The acute D av changes in the ipsilateral hippocampus correlated with the chronic spatial learning and memory impairment evaluated using the Morris water maze ( p < 0.05). Similarly, T 1ρ, T 2 and D av correlated with hippocampal atrophy and with histologically quantified neurodegeneration ( p < 0.01). The early lesion volume and quantitative MRI changes in the perilesional region prefigured the final lesion extent ( p < 0.01). Furthermore, the severity of acute intracerebral hemorrhage correlated with the final cortical atrophy ( p < 0.05), hippocampal atrophy ( p < 0.01), and also with the water maze performance ( p < 0.01). We conclude that, assessment of early quantitative MRI changes in the hippocampus and in the perifocal area may help to predict the long-term outcome after experimental TBI.

Wastewaters from separate chemical factories are treated together in an extended aeration activated sludge plant. The factories produce chemicals for paper industry (e.g. starch), latexes and animal feed. The components of the wastewaters include styrene, tertiary butanol and vinyl acetate. Activated sludge is clarified by sedimentation. During winter time, when the water temperature was 3-12°C, the clarification deteriorated causing carry over of suspended solids containing COD. Enhancement of suspended solids and COD removals was studied in a dissolved air flotation jar test unit. Flotation trials were conducted for activated sludge, sedimentation treated final effluent (tertiary treatment) and separate wastewater fractions. The need for chemicals, flocculation and amount of recycle water were judged according to the achieved removals. Dissolved air flotation was found well suited for the clarification of activated sludge, but not technically and economically feasible for the clarification of the wastewater streams before the activated sludge treatment.

To study macular topography and contrast sensitivity (CS) in diabetic and nondiabetic women during pregnancy and post partum. A prospective study of 46 pregnant women with insulin-dependent diabetes and 11 nondiabetic pregnant controls. Macular surface topography was analyzed by Heidelberg Retinal Tomograph. Volume above the reference plane (VARP) was measured with 1.0-, 1.5-, 2.0-, and 3.0-mm-diameter circles. CS was measured with the Vistech 6500 Contrast Test System. The diabetic women had greater VARP than the controls measured with the 1.5-mm diameter circle. In diabetic women, the mean VARP was 0.084+/-0.064 mm(3) (mean +/- SD) in the first trimester, 0.080 +/- 0.056 mm(3) in the third trimester, and 0.087 +/- 0.067 mm(3) 3 months post partum compared with the values of 0.069+/-0.043, 0.054+/-0.024, and 0.036+/-0.020 mm(3) in the controls ( P=0.036 between groups). In diabetic women requiring laser treatment, the difference from controls was more significant ( P<0.001). CS at 3 cpd and 6.0 cpd was lower in diabetic women than in controls throughout pregnancy and post partum ( P=0.012 and P=0.043). A statistically significant negative correlation appeared between macular topography and CS during the third trimester; between cpd 6 and VARP 1.5 mm ( r=-0.471, P=0.001), and between cpd 6 and VARP 2.0 mm ( r=-0.446, P=0.002). In the diabetic women, especially in those with clear progression of retinopathy during pregnancy and post-partum, the macula seemed to be slightly more elevated than in the controls, and CS was lower at mid-range spatial frequencies. CS loss in the diabetic women correlated with macular elevation during the third trimester.

Wastewaters from separate chemical factories are treated together in an extended aeration activated sludge plant. The factories produce chemicals for paper industry (e.g. starch), latexes and animal feed. The components of the wastewaters include styrene, tertiary butanol and vinyl acetate. Activated sludge is clarified by sedimentation. During winter time, when the water temperature was 3-12 degree C, the clarification deteriorated causing carry over of suspended solids containing COD. Enhancement of suspended solids and COD removals was studied in a dissolved air flotation jar test unit. Flotation trials were conducted for activated sludge, sedimentation treated final effluent (tertiary treatment) and separate wastewater fractions. The need for chemicals, flocculation and amount of recycle water were judged according to the achieved removals. Dissolved air flotation was found well suited for the clarification of activated sludge, but not technically and economically feasible for the clarification of the wastewater streams before the activated sludge treatment.

To find out whether the levels of insulin-like growth factor-I (IGF-I), IGF binding protein-1 (IGFBP-1), highly phosphorylated IGFBP-1 (hpIGFBP-1), and IGF binding protein-3 (IGFBP-3) are related to the progression of diabetic retinopathy (DR) during pregnancy and postpartum. In a prospective study of 42 pregnant women with Type 1 diabetes and 9 nondiabetic controls, DR was graded from fundus photographs. Levels of serum total IGF-I and two different phosphoisoform patterns of IGFBP-1 and IGFBP-3 were measured during the first and third trimester of pregnancy and 3 months postpartum. Both the levels of serum total IGF-I ( P<.0001) and IGFBP-3 ( P=.003) were lower in the diabetic than in the nondiabetic women during pregnancy and postpartum (repeated-measures ANOVA between the groups). Additionally, the IGF-I and IGFBP-3 levels tended to be lower in the diabetic women with more severe DR at baseline than in those with less severe DR. There were no statistically significant differences in the levels of IGF-I and IGFBP-3 in the diabetic women with progression of DR compared with those without. No statistical differences appeared in the IGFBP-1 phosphoisoform patterns between the groups. In diabetic women, mean serum levels of IGF-1 and IGFBP-3 are lower than in nondiabetic controls during pregnancy and/or postpartum. Because there was no clear connection between the IGF system and progression of DR during pregnancy, it is unlikely that these substances mediate the tendency of DR to progress during pregnancy.

ObjectiveAn adequate bowel cleansing is essential for a successful colonoscopy. Although purgative consumption is safe for the patient, there is little consensus on how the intestinal microbiota is affected by the procedure, especially regarding the potential long-term consequences.Design23 healthy subjects were randomised into two study groups consuming a bowel preparation (Moviprep), either in two separate doses of 1 L or as a single 2-L dose. Participants donated faecal samples at the baseline, after bowel cleansing, 14 and 28 days after the treatment. The intestinal microbiota composition was determined with phylogenetic microarray as well as quantitative PCR analysis and correlated with the previously quantified faecal serine proteases.ResultsThe lavage introduced an instant and substantial change to the intestinal microbiota. The total microbial load was decreased by 31-fold and 22% of the participants lost the subject-specificity of their microbiota. While the bacterial levels and community composition were essentially restored within 14 days, the rate of recovery was dose dependent: consumption of the purgative in a single dose had a more severe effect on the microbiota composition than that of a double dose, and notably increased the levels of Proteobacteria, Fusobacteria and bacteria related to Dorea formicigenerans. The abundance of the latter also correlated with the amount of faecal serine proteases that were increased after purging.ConclusionsOur results suggest that the bowel cleansing using two separate dosages introduces fewer alterations to the intestinal microbiota than a single dose and hence may be preferred in clinical practice.

HIV and SIV infection dynamics are commonly investigated by measuring plasma viral loads. However, this total viral load value represents the sum of many individual infection events, which are difficult to independently track using conventional sequencing approaches. To overcome this challenge, we generated a genetically tagged virus stock (SIVmac239M) with a 34-base genetic barcode inserted between the vpx and vpr accessory genes of the infectious molecular clone SIVmac239. Next-generation sequencing of the virus stock identified at least 9,336 individual barcodes, or clonotypes, with an average genetic distance of 7 bases between any two barcodes. In vitro infection of rhesus CD4+ T cells and in vivo infection of rhesus macaques revealed levels of viral replication of SIVmac239M comparable to parental SIVmac239. After intravenous inoculation of 2.2x105 infectious units of SIVmac239M, an average of 1,247 barcodes were identified during acute infection in 26 infected rhesus macaques. Of the barcodes identified in the stock, at least 85.6% actively replicated in at least one animal, and on average each barcode was found in 5 monkeys. Four infected animals were treated with combination antiretroviral therapy (cART) for 82 days starting on day 6 post-infection (study 1). Plasma viremia was reduced from >106 to <15 vRNA copies/mL by the time treatment was interrupted. Virus rapidly rebounded following treatment interruption and between 87 and 136 distinct clonotypes were detected in plasma at peak rebound viremia. This study confirmed that SIVmac239M viremia could be successfully curtailed with cART, and that upon cART discontinuation, rebounding viral variants could be identified and quantified. An additional 6 animals infected with SIVmac239M were treated with cART beginning on day 4 post-infection for 305, 374, or 482 days (study 2). Upon treatment interruption, between 4 and 8 distinct viral clonotypes were detected in each animal at peak rebound viremia. The relative proportions of the rebounding viral clonotypes, spanning a range of 5 logs, were largely preserved over time for each animal. The viral growth rate during recrudescence and the relative abundance of each rebounding clonotype were used to estimate the average frequency of reactivation per animal. Using these parameters, reactivation frequencies were calculated and ranged from 0.33-0.70 events per day, likely representing reactivation from long-lived latently infected cells. The use of SIVmac239M therefore provides a powerful tool to investigate SIV latency and the frequency of viral reactivation after treatment interruption.