Human Synaptic Plasticity Gene Expression Profile and Dendritic Spine Density Changes in HIV-Infected Human CNS Cells: Role in HIV-Associated Neurocognitive Disorders (HAND)

HIV-associated neurocognitive disorders (HAND) is characterized by development of cognitive, behavioral and motor abnormalities, and occur in approximately 50% of HIV infected individuals. Our current understanding of HAND emanates mainly from HIV-1 subtype B (clade B), which is prevalent in USA and Western countries. However very little information is available on neuropathogenesis of HIV-1 subtype C (clade C) that exists in Sub-Saharan Africa and Asia. Therefore, studies to identify specific neuropathogenic mechanisms associated with HAND are worth pursuing to dissect the mechanisms underlying this modulation and to prevent HAND particularly in clade B infection. In this study, we have investigated 84 key human synaptic plasticity genes differential expression profile in clade B and clade C infected primary human astrocytes by using RT2 Profile PCR Array human Synaptic Plasticity kit. Among these, 31 and 21 synaptic genes were significantly (≥3 fold) down-regulated and 5 genes were significantly (≥3 fold) up-regulated in clade B and clade C infected cells, respectively compared to the uninfected control astrocytes. In flow-cytometry analysis, down-regulation of postsynaptic density and dendrite spine morphology regulatory proteins (ARC, NMDAR1 and GRM1) was confirmed in both clade B and C infected primary human astrocytes and SK-N-MC neuroblastoma cells. Further, spine density and dendrite morphology changes by confocal microscopic analysis indicates significantly decreased spine density, loss of spines and decreased dendrite diameter, total dendrite and spine area in clade B infected SK-N-MC neuroblastoma cells compared to uninfected and clade C infected cells. We have also observed that, in clade B infected astrocytes, induction of apoptosis was significantly higher than in the clade C infected astrocytes. In conclusion, this study suggests that down-regulation of synaptic plasticity genes, decreased dendritic spine density and induction of apoptosis in astrocytes may contribute to the severe neuropathogenesis in clade B infection.

Modeling Nymphoides architecture: A morphological analysis of Nymphoides aquatica (Menyanthaceae)

• Premise of the study: Species in the aquatic genus Nymphoides have inflorescences that appear to arise from the petioles of floating leaves. The inflorescence-floating leaf complex can produce vegetative propagules and/or additional inflorescences and leaves. We analyzed the morphology of N. aquatica to determine how this complex relates to whole plant architecture and whether whole plant growth is sympodial or monopodial. • Methods: We used dissections, measurements, and microscopic observations of field-collected plants and plants cultivated for 2 years in outdoor tanks in south Florida, USA. • Key results: Nymphoides aquatica had a submerged plagiotropic rhizome that produced floating leaves in an alternate/spiral phyllotaxy. Rhizomes were composed of successive sympodial units that varied in the number of leaves produced before the apex terminated. The basic sympodial unit had a prophyll that subtended a renewal-shoot bud, a short-petioled leaf (SPL) with floating lamina, and an inflorescence; the SPL axillary bud expanded as a vegetative propagule. Plants produced either successive basic sympodial units or expanded sympodia that intercalated long-petioled leaves between the prophyll and the SPL. • Conclusions: Nymphoides aquatica grows sympodially, forming a rhizome composed of successive basic sympodia and expanded sympodial units. Variations on these types of sympodial growth help explain the branching patterns and leaf morphologies described for other Nymphoides species. Monitoring how these two sympodial phases are affected by water depth provides an ecologically meaningful way to assess N. aquatica’s responses to altered hydrology.

Fractal analysis of flocs formed in edible oil -water emulsions by image analysis techniques

Edible oil is an important contaminant in water and wastewater. Oil droplets smaller than 40 μm may remain in effluent as an emulsion and combine with other contaminants in water. Coagulation/flocculation processes are used to remove oil droplets from water and wastewater. By adding a polymer at proper dose, small oil droplets can be flocculated and separated from water. The purpose of this study was to characterize and analyze the morphology of flocs and floc formation in edible oil-water emulsions by using microscopic image analysis techniques. The fractal dimension, concentration of polymer, effect of pH and temperature are investigated and analyzed to develop a fractal model of the flocs. Three types of edible oil (corn, olive, and sunflower oil) at concentrations of 600 ppm (by volume) were used to determine the optimum polymer dosage and effect of pH and temperature. To find the optimum polymer dose, polymer was added to the oil-water emulsions at concentration of 0.5, 1.0, 1.5, 2.0, 3.0 and 3.5 ppm (by volume). The clearest supernatants obtained from flocculation of corn, olive, and sunflower oil were achieved at polymer dosage of 3.0 ppm producing turbidities of 4.52, 12.90, and 13.10 NTU, respectively. This concentration of polymer was subsequently used to study the effect of pH and temperature on flocculation. The effect of pH was studied at pH 5, 7, 9, and 11 at 30°C. Microscopic image analysis was used to investigate the morphology of flocs in terms of fractal dimension, radius of oil droplets trapped in floc, floc size, and histograms of oil droplet distribution. Fractal dimension indicates the density of oil droplets captured in flocs. By comparison of fractal dimensions, pH was found to be one of the most important factors controlling droplet flocculation. Neutral pH or pH 7 showed the highest degree of flocculation, while acidic (pH 5) and basic pH (pH 9 and pH 11) showed low efficiency of flocculation. The fractal dimensions achieved from flocculation of corn, olive, and sunflower oil at pH 7 and temperature 30°C were 1.2763, 1.3592, and 1.4413, respectively. The effect of temperature was explored at temperatures 20°, 30°, and 40°C and pH 7. The results of flocculation of oil at pH 7 and different temperatures revealed that temperature significantly affected flocculation. The fractal dimension of flocs formed in corn, olive and sunflower oil emulsion at pH 7 and temperature 20°, 30°, and 40°C were 1.82, 1.28, 1.29, 1.62, 1.36, 1.42, 1.36, 1.44, and 1.28, respectively. After comparison of fractal dimension, radius of oil droplets captured, and floc length in each oil type, the optimal flocculation temperature was determined to be 30°C. ^