Purification and active site modification studies on glyoxalase I from monkey intestinal mucosa

Glyoxalase I ((R)-S-lactoylglutathione methylglyoxal-lyase (isomerizing), EC 4.4.1.5) from monkey intestinal mucosa was purified to homogeneity. The purified enzyme had a molecular weight of 48,000, composed of two apparently identical subunits. Active-site modification was carried out on the purified enzyme in presence and absence of S-hexylglutathione, a reversible competitive inhibitor of glyoxalase I. Modification by tetranitromethane and N-acetylimidazole caused inactivation of the enzyme. Inactivation by N-acetylimidazole was reversible with hydroxylamine treatment, suggesting the importance of tyrosine residues for the activity of the enzyme. The enzyme was inactivated by 2-hydroxy-5-nitrobenzyl bromide, N-bromosuccinimide, 2,4,6-trinitrobenzenesulphonic acid, pyridoxal phosphate and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, indicating the importance of tryptophan, lysine and glutamic acid/aspartic acid residues for the activity of the enzyme. The enzyme was inactivated by diethyl pyrocarbonate and the activity was not restored by hydroxylamine treatment, suggesting that histidine residues may not be important for activity. Modification by N-ethylmaleimide and p-hydroxymercuribenzoate did not affect its activity, indicating that sulphydryl groups may not be important for activity. These studies indicated that the amino acids present in the active site of glyoxalase I from intestinal mucosa which may be important for activity are tyrosine, tryptophan, lysine and glutamic acid/aspartic acid residues.     

Verification of a virtual fields method to extract the mechanical properties of human optic nerve head tissues in vivo

We aimed to verify a custom virtual fields method (VFM) to estimate the patient-specific biomechanical properties of human optic nerve head (ONH) tissues, given their full-field deformations induced by intraocular pressure (IOP). To verify the accuracy of VFM, we first generated ‘artificial’ ONH displacements from predetermined (known) ONH tissue biomechanical properties using finite element analysis. Using such deformations, if we are able to match back the known biomechanical properties, it would indicate that our VFM technique is accurate. The peripapillary sclera was assumed anisotropic hyperelastic, while all other ONH tissues were considered isotropic. The simulated ONH displacements were fed into the VFM algorithm to extract back the biomechanical properties. The robustness of VFM was also tested against rigid body motions and noise added to the simulated displacements. Then, the computational speed of VFM was compared to that of a gold-standard stiffness measurement method (inverse finite element method or IFEM). Finally, as proof of principle, VFM was applied to IOP-induced ONH deformation data (obtained from one subject’s eye imaged with OCT), and the biomechanical properties of the prelamina and lamina cribrosa (LC) were extracted. From given ONH displacements, VFM successfully matched back the biomechanical properties of ONH tissues with high accuracy and efficiency. For all parameters, the percentage errors were less than 0.05%. Our method was insensitive to rigid body motions and was also able to recover the material parameters in the presence of noise. VFM was also found 125 times faster than the gold-standard IFEM. Finally, the estimated shear modulus for the prelamina and the LC of the studied subject’s eye were 33.7 and 63.5 kPa, respectively. VFM may be capable of measuring the biomechanical properties of ONH tissues with high speed and accuracy. It has potential in identifying patient-specific ONH biomechanical properties in the clinic if combined with optical coherence tomography.     

16 S rRNA gene diversity and gut microbial composition of the Indian white shrimp (Penaeus indicus)

Antonie van Leeuwenhoek - The endemic Indian white shrimp (Penaeus indicus) is an economically important crustacean species, distributed in the Indo-West Pacific region. Knowledge of its gut...     

Psoralen production in hairy roots and adventitious roots cultures of Psoralea coryfolia

Psoralea corylifolia is an endangered plant producing various compounds of medical importance. Adventitious roots and hairy roots were induced in cultures prepared from hypocotyl explants. Psoralen content was evaluated in both root types grown either in suspension cultures or on agar solidified medium. Psoralen content was ~3 mg g⁻¹ DW in suspension grown hairy roots being higher than in solid grown hairy roots and in solid and suspension-grown adventitious roots.     

Comparative effects of β-carotene and fucoxanthin on retinol deficiency induced oxidative stress in rats

This study aimed at comparing antioxidant potential of fucoxanthin (FUCO) with β-carotene in relieving lipid peroxidation (Lpx) caused by retinol deficiency (RD) in rats. RD rats (n = 45) were fed a dose of either β-carotene (0.81 μmol) or FUCO (0.83 μmol). Plasma and liver lipid peroxide levels and activity of antioxidant enzymes catalase (CAT) and glutathione transferase (GST) were measured for 8 h. Results revealed that RD increased (P < 0.05) Lpx in plasma and liver by 34.3% and 19.4%, while the CAT activity in plasma (89%) and liver microsomes (91%) and GST in liver homogenate (31%) and liver microsomes (30%) were decreased (P < 0.05) compared to control (rats fed basal diet). FUCO suppressed (P < 0.05) the Lpx level by 7–85% (plasma) and 24–72% (liver) as compared to β-carotene (51–76%, 33–65%) over a period of 8 h. The activity of CAT in plasma and liver microsomes was higher (P < 0.05) in FUCO (90–95%, 85–93%) and β-carotene (87–96%, 79–91%) groups as compared to RD group. Similarly, the activity of GST in liver and its microsomes was also elevated (P < 0.05) in FUCO (44–51%, 22–51%) and β-carotene (19–54%, 30–43%) groups as compared to RD group. Results demonstrate that FUCO has greater potential than β-carotene in modulating Lpx, CAT, GST in plasma and liver of RD rats.     

MMR/c-Abl-dependent activation of ING2/p73α signaling regulates the cell death response to N-methyl-N′-nitro-N-nitrosoguanidine

Agents inducing O⁶-methylguanine (O⁶MeG) in DNA such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are cytotoxic and a deficiency in mismatch repair (MMR) results in lack of sensitivity to this genotoxin (termed alkylation tolerance). Here, we show that ING2, a member of the inhibitor of growth family, is required for cell death induced by MNNG. We further observe that MNNG treatment increases cellular protein levels of ING2 that is dependent on intact MMR function and that MNNG-induced ING2 localizes and associates with p73α in the nucleus. Suppression of ING2 by short hairpin RNA (shRNA) in MMR-proficient colorectal cancer cells decreased its sensitivity to MNNG and, in addition, abrogated MNNG-induced stabilization and acetylation of p73α. Interestingly, suppression of p73α had a greater impact on MNNG-induced cell death than ING2 leading us to conclude that ING2 regulates the cell death response, in part, through p73α. Inhibition of c-Abl by STI571 or suppression of c-Abl expression by shRNA blocked ING2 induction and p73α acetylation induced by this alkylator. Similarly, suppression of MMR (MLH1) by shRNA abrogated ING2 induction/p73α acetylation. Taken together, these results demonstrate that MLH1/c-Abl-dependent activation of ING2 > p73α signaling regulates cell death triggered by MNNG and further suggests that dysregulation of this event may, in part, be responsible for alkylation tolerance observed in MMR compromised cells.     

Planning Transboundary Ecological Risk Assessments at Military Installations

Ecological risk assessments at military installations that are performed to support natural resources management objectives rely on information from the surrounding region. Stressors such as noise, ozone, and ozone precursors cross installation boundaries, and effects of urbanization and highway development are regional in scale. Ecological populations are not limited to one side of the installation boundary. Therefore, a framework for transboundary ecological risk assessment at military installations is under development. This article summarizes the problem formulation stage. Components include: (1) regional management goals such as installation Integrated Natural Resources Management Plans and land acquisition, (2) involvement of multiple stressors, and (3) large-scale assessment endpoint entities. Challenges of selecting measures of exposure include: quantifying exposure to aggregate stressors, describing land cover consistently in the region, describing rates of land-cover transition, scaling local measurements to a region, and aggregating or isolating exposures from within and outside of the installation. Measures of effect that are important to transboundary or regional ecological risk assessments at military installations are those that represent: effects at a distance from the stressor, large-scale effects, effects of habitat change or fragmentation, spatial extrapolations of localized effects, and integrated effects of multiple stressors. These factors are reflected in conceptual models.     

Problems in seawater industrial desalination processes and potential sustainable solutions: a review

Seawater desalination has significantly developed towards membrane technology than phase change process during last decade. Seawater reverse osmosis (SWRO) in general is the most familiar process due to higher water recovery and lower energy consumption compared to other available desalination processes. Despite major advancements in SWRO technology, desalination industry is still facing significant amount of practical issues. Therefore, the potentials and problems faced by current SWRO industries and essential study areas are discussed in this review for the benefit of desalination industry. It is important to consider all the following five components in SWRO process i.e. (1) intake (2) pre-treatment (3) high pressure pumping (4) membrane separation (performance of membranes and brine disposal) and (5) product quality. Development of higher corrosion resistant piping materials or coating materials, valves, and pumps is believed to be in higher research demand. Furthermore, brine management, that includes brine disposal and resource recovery need further attention. Pre-treatment sludge management and reduced cleaning in place flush volume will reduce the capital costs associated with evaporation ponds and the maintenance costs associated with disposal and transportation reducing the unit cost of water.