Cytogenetic studies of chromium (III) oxide nanoparticles on Allium cepa root tip cells.

The current study evaluates the cytogenetic effects of chromium (III) oxide nanoparticles on the root cells of Allium cepa. The root tip cells of A. cepa were treated with the aqueous dispersions of Cr2O3 nanoparticles (NPs) at five different concentrations (0.01, 0.1, 1, 10, and 100µg/mL) for 4hr. The colloidal stability of the nanoparticle suspensions during the exposure period were ascertained by particle size analyses. After 4hr exposure to Cr2O3 NPs, a significant decrease in mitotic index (MI) from 35.56% (Control) to 35.26% (0.01µg/mL), 34.64% (0.1µg/mL), 32.73% (1µg/mL), 29.6% (10µg/mL) and 20.92% (100µg/mL) was noted. The optical, fluorescence and confocal laser scanning microscopic analyses demonstrated specific chromosomal aberrations such as-chromosome stickiness, chromosome breaks, laggard chromosome, clumped chromosome, multipolar phases, nuclear notch, and nuclear bud at different exposure concentrations. The concentration-dependent internalization/bio-uptake of Cr2O3 NPs may have contributed to the enhanced production of anti oxidant enzyme, superoxide dismutase to counteract the oxidative stress, which in turn resulted in observed chromosomal aberrations and cytogenetic effects. These results suggest that A. cepa root tip assay can be successfully applied for evaluating environmental risk of Cr2O3 NPs over a wide range of concentrations.

Combined toxicity of two crystalline phases (anatase and rutile) of Titania nanoparticles towards freshwater microalgae: Chlorella sp.

In view of the increasing usage of anatase and rutile crystalline phases of titania NPs in the consumer products, their entry into the aquatic environment may pose a serious risk to the ecosystem. In the present study, the possible toxic impact of anatase and rutile nanoparticles (individually and in binary mixture) was investigated using freshwater microalgae, Chlorella sp. at low exposure concentrations (0.25, 0.5 and 1mg/L) in freshwater medium under UV irradiation. Reduction of cell viability as well as a reduction in chlorophyll content were observed due to the presence of NPs. An antagonistic effect was noted at certain concentrations of binary mixture such as (0.25, 0.25), (0.25, 0.5), and (0.5, 0.5) mg/L, and an additive effect for the other combinations, (0.25, 1), (0.5, 0.25), (0.5, 1), (1, 0.25), (1, 0.5), and (1, 1) mg/L. The hydrodynamic size analyses in the test medium revealed that rutile NPs were more stable in lake water than the anatase and binary mixtures [at 6h, the sizes of anatase (1mg/L), rutile NPs (1mg/L), and binary mixture (1, 1mg/L) were 948.83±35.01nm, 555.74±19.93nm, and 1620.24±237.87nm, respectively]. The generation of oxidative stress was found to be strongly dependent on the crystallinity of the nanoparticles. The transmission electron microscopic images revealed damages in the nucleus and cell membrane of algal cells due to the interaction of anatase NPs, whereas rutile NPs were found to cause chloroplast and internal organelle damages. Mis-shaped chloroplasts, lack of nucleus, and starch-pyrenoid complex were noted in binary-treated cells. The findings from the current study may facilitate the environmental risk assessment of titania NPs in an aquatic ecosystem.

Cytotoxicity of TiO2 nanoparticles towards freshwater sediment microorganisms at low exposure concentrations.

There is a persistent need to assess the effects of TiO2 nanoparticles on the aquatic ecosystem owing to their increasing usage in consumer products and risk of environmental release. The current study is focused on TiO2 nanoparticle-induced acute toxicity at sub-ppm level (≤1ppm) on the three different freshwater sediment bacterial isolates and their consortium under two different irradiation (visible light and dark) conditions. The consortium of the bacterial isolates was found to be less affected by the exposure to the nanoparticles compared to the individual cells. The oxidative stress contributed considerably towards the cytotoxicity under both light and dark conditions. A statistically significant increase in membrane permeability was noted under the dark conditions as compared to the light conditions. The optical and fluorescence microscopic images showed aggregation and chain formation of the bacterial cells, when exposed to the nanoparticles. The electron microscopic (SEM, TEM) observations suggested considerable damage of cells and bio-uptake of nanoparticles. The exopolysaccrides (EPS) production and biofilm formation were noted to increase in the presence of the nanoparticles, and expression of the key genes involved in biofilm formation was studied by RT-PCR.

Kinetic behaviour of calf intestinal alkaline phosphatase with pNPP.

The hydrolysis of p-nitrophenyl phosphate (pNPP) by calf intestinal alkaline phosphatase (CIAP) was investigated with respect to kinetic parameters such as V(max), K(m) and K(cat) under varying pH, buffers, substrate concentration, temperature and period of incubation. Highest activity was obtained with Tris-HCl at pH 11, while in the case of glycine-NaOH buffer the peak activity was recorded at pH 9.5. The enzyme showed the following kinetic characteristics with pNPP in 50 mM Tris-HCl at pH 11 and 100 mM glycine-NaOH at pH 9.5 at an incubation temperature of 37 degrees C: V(max), 3.12 and 1.6 micromoles min(-1) unit(-1); K(m), 7.6 x 10(-4) M and 4 x 10(-4) M; and K(cat), 82.98 s(-1) and 42.55 s(-1), respectively. CIAP displayed a high temperature optimum of 45 degrees C at pH 11. The kinetic behaviour of the enzyme under different parameters suggested that the enzyme might undergo subtle conformational changes in response to the buffers displaying unique characteristics. Bioprecipitation of Cu2+ from 50 ppm of CuCl2 solution was studied where 64.3% of precipitation was obtained. P(i) generated from CIAP-mediated hydrolysis of pNPP was found to bind with copper and precipitated as copper-phosphate. Thus, CIAP could be used as a test candidate in bioremediation of heavy metals from industrial wastes through generation of metal-phosphate complexes.

Enzymatically mediated bioprecipitation of heavy metals from industrial wastes and single ion solutions by mammalian alkaline phosphatase.

The study was aimed at investigating the potential use of calf intestinal alkaline phosphatase (CIAP) enzyme in the removal of heavy metals (Cd(2+), Ni(2+), Co(2+) and Cr(3+/6+)) from single ion solutions as well as tannery and electroplating effluents. CIAP mediated bioremediation (white biotechnology) is a novel technique that is eco-friendly and cost effective unlike the conventional chemical technologies. Typical reactions containing the enzyme (CIAP) and p-nitrophenyl phosphate (pNPP) as substrate in Tris-HCl buffer (pH 8 and 11) and either single ion metal solutions (250 ppm and 1000 ppm) or effluents from tannery or electroplating industry were incubated at 37°C for 30 min, 60 min and 120 min. The inorganic phosphate (P(i)) generated due to catalytic breakdown of pNPP complexes free metal ions as metal-phosphate and the amount of metal precipitated was derived by estimating the reduction in the free metal ion present in the supernatant of reactions employing atomic absorption spectrophotometer (AAS). Better precipitation of metal was obtained at pH 11 than at pH 8 and between the two concentrations of different metals tested, an initial metal concentration of 250 ppm in the reaction gave more precipitation than with 1000 ppm. Experimental data showed that at pH 11, the percentage of removal of metal ions (for an initial concentration of 250 ppm) was in the following order: Cd(2+) (80.99%) > Ni(2+) (64.78%) > Cr(3+) > (46.15%) > Co(2+) (36.47%) > Cr(6+) (32.33%). The overall removal of Cr(3+) and Cr(6+) from tannery effluent was 32.77% and 37.39% respectively in 120 min at pH 11. Likewise, the overall removal of Cd(2+), Co(2+) and Ni(2+) from electroplating effluent was 50.42%, 13.93% and 38.64% respectively in 120 min at pH 11. The study demonstrates that bioprecipitation by CIAP may be a viable and environmental friendly method for clean-up of heavy metals from tannery and electroplating effluents.

Compelled body weight shift approach in rehabilitation of individuals with chronic stroke.

PURPOSE: This study was designed to evaluate the effectiveness of the compelled body weight shift (CBWS) therapy approach in the rehabilitation of individuals with chronic stroke. CBWS involves a forced shift of body weight toward a person's affected side by means of a shoe insert that establishes a lift of the nonaffected lower extremity. METHOD: Eighteen individuals with chronic, unilateral stroke (mean age 57.7 ± 11.9 years, with a range of 35-75 years; mean time since stroke 6.7 ± 3.9 years, with a range of 1.1-14.1 years) who showed asymmetrical stance were randomly divided into 2 groups: the experimental group received 6 weeks of physical therapy combined with CBWS therapy, and the control group received only physical therapy. Both groups underwent a battery of identical tests (Fugl-Meyer Assessment, Berg Balance Scale, weight bearing, and gait velocity) before the start of the rehabilitation intervention, following its completion, and 3 months after the end of therapy. RESULTS: After the intervention, weight bearing on the affected side (measured with the Balance Master) increased in the experimental group to a larger degree compared to the control group (9.7% vs 6.4%). Similarly, gait velocity increased 10.5% in the experimental group compared to the control group. Improvements in weight bearing and gait velocity were maintained in the experimental group after the 3-month retention period. CONCLUSION: The study outcome revealed that a 6-week intervention involving CBWS therapy could result in a long-lasting improvement of the symmetry of weight bearing and velocity of gait in individuals with chronic stroke.

Gait assessment during the initial fitting of an ankle foot orthosis in individuals with stroke.

PURPOSE: To evaluate if the measurement of gait parameters, examined during the fitting of an Ankle Foot Orthosis (AFO), has a beneficial effect on the gait pattern of individuals who were affected by a stroke. Also, this study seeks to provide evidence regarding the use of the portable GaitRite system in a clinical setting. METHOD: Before-after trial conducted at a stroke outpatient orthotic clinic of a freestanding rehabilitation hospital. Thirteen individuals with acute and 27 individuals with chronic stroke participated in the assessment of gait velocity, cadence, step length and stance phase. RESULTS: AFO use significantly improves gait velocity, cadence, step, and stride length in individuals with hemiparesis due to stroke. CONCLUSION: The results of the current study indicate that the assessment of temporo-spatial characteristics of gait can be incorporated into a clinical routine. This will be useful for patient education, justification of medical necessity of paying, monitoring progress, and in the decision-making process of weaning patients off orthoses.

Cardiovascular effects of the supraglottic and super-supraglottic swallowing maneuvers in stroke patients with dysphagia.

The prolonged voluntary closure of the glottis during the supraglottic and super-supraglottic swallowing techniques may create the Valsalva maneuver. The Valsalva maneuver has been associated with sudden cardiac death and cardiac arrhythmias. This study describes the effects of the supraglottic and super-supraglottic swallowing techniques on the cardiovascular system. Subjects included 23 patients from an acute inpatient rehabilitation hospital. Subject groups included recent stroke, dysphagia. and a history of coronary artery disease (Group 1, N = 11). recent stroke and dysphagia with no known coronary artery disease (Group 2, N = 4), and orthopedic diagnosis with no known dysphagia or coronary artery disease (Group 3, N = 8). Cardiac status was monitored for 4 hours during swallowing training, regular therapy sessions, and a meal. For Groups I and 2, 86.6% (13 out of 15) of the subjects demonstrated abnormal cardiac findings during the swallowing session including supraventricular tachycardia, premature atrial contractions, and premature ventricular contractions. Arrhythmia subsided within a few minutes after the session and did not occur during other activities. In Group 3 (control group), none of the subjects demonstrated abnormal cardiac findings except for bradycardia in one subject. It is suggested that the supraglottic and super-supraglottic swallow maneuvers may be contraindicated for patients with a history of stroke or coronary artery disease.