Structural and functional analysis of two universal stress proteins YdaA and YnaF from Salmonella typhimurium: possible roles in microbial stress tolerance

In many organisms ``Universal Stress Proteins'' CUSPS) are induced in response to a variety of environmental stresses. Here we report the structures of two USPs, YnaF and YdaA from Salmonella typhimurium determined at 1.8 angstrom and 2.4 angstrom resolutions, respectively. YnaF consists of a single USP domain and forms a tetrameric organization stabilized by interactions mediated through chloride ions. YdaA is a larger protein consisting of two tandem USP domains. Two protomers of YdaA associate to form a structure similar to the YnaF tetramer. YdaA showed ATPase activity and an ATP binding motif G-2X-G-9X-G(S/T/N) was found in its C-terminal domain. The residues corresponding to this motif were not conserved in YnaF although YnaF could bind ATP. However, unlike YdaA, YnaF did not hydrolyse ATP in vitro. Disruption of interactions mediated through chloride ions by selected mutations converted YnaF into an ATPase. Residues that might be important for ATP hydrolysis could be identified by comparing the active sites of native and mutant structures. Only the C-terminal domain of YdaA appears to be involved in ATP hydrolysis. The structurally similar N-terminal domain was found to bind a zinc ion near the segment equivalent to the phosphate binding loop of the C-terminal domain. Mass spectrometric analysis showed that YdaA might bind a ligand of approximate molecular weight 800 daltons. Structural comparisons suggest that the ligand, probably related to an intermediate in lipid A biosynthesis, might bind at a site close to the zinc ion. Therefore, the N-terminal domain of YdaA binds zinc and might play a role in lipid metabolism. Thus, USPs appear to perform several distinct functions such as ATP hydrolysis, altering membrane properties and chloride sensing. (C) 2015 Elsevier Inc. All rights reserved.

Influence of initial degree of saturation on swell pressures of compacted Barmer bentonite specimens

Densely compacted bentonite or bentonite-sand mixture has been identified as suitable buffer in deep geological repositories as its exceptionally high swelling capacity enables tight contact between the waste canister and surrounding rock. The degree of saturation of the compacted bentonite buffer can increase upon ingress of groundwater from the surrounding rock mass or decrease from evaporation due to high temperature (50-210 degrees C) derived from the waste canister. Available studies indicate that the influence of initial moisture content or degree of saturation on the swell pressure or swell potential of compacted bentonites is unclear. Some studies suggest that initial degree of saturation has an influence, while others suggest that it does not have bearing on the swell pressure of compacted bentonites. This paper examines the influence of initial degree of saturation in montmorillonite voids (termed,S-r,S-MF) on swell pressure of compacted Barmer bentonite-sand mixtures (dry density range: 1.4-2 Mg/m(3)) from micro-structural considerations. The experimental results bring out that, constant dry density specimens that developed similar number of hydration layers upon wetting developed comparable swell pressures and were unaffected by variations in initial S-r,S-MF values. Comparatively, constant dry density specimens that developed dis-similar number of hydration layers upon wetting established different swell pressures and were responsive to variations in initial S-r,S-MF. (C) 2015 Elsevier Ltd. All rights reserved.

Role of synergistic pi-pi stacking and X-H center dot center dot center dot Cl (X = C, N, O) H-bonding interactions in gelation and gel phase crystallization

The self-assembly of p-pyridyl-ended oligo-p-phenylenevinylenes (OPVs) in ethanol leads to the formation of either hollow or solid microrods. The corresponding protonated OPVs with n-butyl chains induce transparent gelation and also gel phase crystallization owing to various synergistic noncovalent interactions. The chloride ion-selective gelation, AIEE and stimuli responsiveness of the gel are also observed.

Experimental and numerical studies on protection of buried pipelines and underground utilities using geocells

This paper presents the results of the laboratory model tests and the numerical studies conducted on small diameter PVC pipes, buried in geocell reinforced sand beds. The aim of the study was to evaluate the suitability of the geocell reinforcement in protecting the underground utilities and buried pipelines. In addition to geocells, the efficacy of only geogrid and geocell with additional basal geogrid cases were also studied. A PVC (Poly Vinyl Chloride) pipe with external diameter 75 mm and thickness 1.4 mm was used in the experiments. The vehicle tire contact pressure was simulated by applying the pressure on the top of the bed with the help of a steel plate. Results suggest that the use of geocells with additional basal geogrid considerably reduces the deformation of the pipe as compared to other types of reinforcements. Further, the depth of placement of pipe was also varied between 1B to 2B (B is the width of loading plate) below the plate in the presence of geocell with additional basal geogrid. More than 50% reduction in the pressure and more than 40% reduction in the strain values were observed in the presence of reinforcements at different depths as compared to the unreinforced beds. Conversely, the performance of the subgrade soil was also found to be marginally influenced by the position of the pipe, even in the presence of the relatively stiff reinforcement system. Further, experimental results were validated with 3-dimensional numerical studies using FLAC(3D) (Fast Lagrangian Analysis of Continua in 3D). A good agreement in the measured pipe stain values were observed between the experimental and numerical studies. Numerical studies revealed that the geocells distribute the stresses in the lateral direction and thus reduce the pressure on the pipe. In addition, the results of the 1-g model tests were scaled up to the prototype case of the shallow buried pipeline below the pavement using the appropriate scaling laws. (C) 2015 Elsevier Ltd. All rights reserved.

Gold catalyzed intramolecular hydroalkoxylation assisted ring opening of furans to the corresponding saturated gamma-keto esters

A facile ring opening of furans in furyl propargyl alcohols to the corresponding saturated gamma-keto esters is observed in the gold(III) chloride catalyzed reaction with MeOH. It is found that the ring opening of furan is driven by the intramolecular hydroalkoxylation. Mitigating the intramolecular hydroalkoxylation led to the expected conjugated enyne resulting from the dehydration. (C) 2015 Elsevier Ltd. All rights reserved.

Endoplasmic reticulum targeting tumour selective photocytotoxic oxovanadium(IV) complexes having vitamin-B6 and acridinyl moieties

Oxovanadium(IV) complexes of vitamin-B6 Schiff base, viz., VO(HL1/L-2/L-3)(B)] Cl (1-4), where B is 2,2'-bipyridine (bpy in 1 and 2), 11-(9-acridinyl)dipyrido3,2-a:2',3'-c]phenazine (acdppz in 3 and 4), H2L1 center dot HCl is 3-hydroxy-5-(hydroxymethyl)-4-(((2-hydroxyphenyl)imino)methyl)-2-methylp yridin-1-ium chloride (in 1 and 4), HL2 is 2-(((2-(1H-imidazol-4-yl)ethyl) imino)methyl) phenol (in 2) and HL3 is 4-(((2-(1H-imidazol-4- yl)ethyl)imino)methyl)-5-(hydroxymethyl)-2-methylpyridin-3-ol (in 3) were synthesized, characterized and their cellular uptake, photo-activated cytotoxicity and intracellular localization were studied. Complexes 1a, as the perchlorate salt of 1, and 2a, as the hexafluorophosphate salt of 2, were structurally characterized. Vitamin-B6 transporting membrane carrier (VTC) mediated entry into tumour cells in preference to the normal ones seems to be responsible for the higher cellular uptake of the complexes into HeLa and MCF-7 cells over MCF-10A cells. Complexes 3 and 4 having acdppz as the photosensitizer exhibit remarkable photocytotoxicity in these cancer cells giving IC50 of < 0.9 mu M. The complexes remain non-toxic in the dark. The complexes show photo-induced apoptotic cell death via singlet oxygen (O-1(2)) generation. Fluorescence microscopy reveals specific localization of complex 4 to endoplasmic reticulum (ER) and generation of O-1(2) possibly leads to apoptotic cell death by triggering ER stress response (ERSR).

Endoplasmic reticulum targeting tumour selective photocytotoxic oxovanadium(IV) complexes having vitamin-B6 and acridinyl moieties

Oxovanadium(IV) complexes of vitamin-B6 Schiff base, viz., VO(HL1/L-2/L-3)(B)] Cl (1-4), where B is 2,2'-bipyridine (bpy in 1 and 2), 11-(9-acridinyl)dipyrido3,2-a:2',3'-c]phenazine (acdppz in 3 and 4), H2L1 center dot HCl is 3-hydroxy-5-(hydroxymethyl)-4-(((2-hydroxyphenyl)imino)methyl)-2-methylp yridin-1-ium chloride (in 1 and 4), HL2 is 2-(((2-(1H-imidazol-4-yl)ethyl) imino)methyl) phenol (in 2) and HL3 is 4-(((2-(1H-imidazol-4- yl)ethyl)imino)methyl)-5-(hydroxymethyl)-2-methylpyridin-3-ol (in 3) were synthesized, characterized and their cellular uptake, photo-activated cytotoxicity and intracellular localization were studied. Complexes 1a, as the perchlorate salt of 1, and 2a, as the hexafluorophosphate salt of 2, were structurally characterized. Vitamin-B6 transporting membrane carrier (VTC) mediated entry into tumour cells in preference to the normal ones seems to be responsible for the higher cellular uptake of the complexes into HeLa and MCF-7 cells over MCF-10A cells. Complexes 3 and 4 having acdppz as the photosensitizer exhibit remarkable photocytotoxicity in these cancer cells giving IC50 of < 0.9 mu M. The complexes remain non-toxic in the dark. The complexes show photo-induced apoptotic cell death via singlet oxygen (O-1(2)) generation. Fluorescence microscopy reveals specific localization of complex 4 to endoplasmic reticulum (ER) and generation of O-1(2) possibly leads to apoptotic cell death by triggering ER stress response (ERSR).

Studies on Carbon Mediated Paste Screen Printed Sensors for Blood Glucose Sensing Application

Ready-to-use screen printed glucose sensors are fabricated using Prussian Blue (PB) and Cobalt Phthalocyanine (CoPC) mediated carbon inks as working electrodes. The reference and counter electrodes are screen printed using silver/silver chloride and graphitic carbon paste respectively. The screen printed reference electrodes (internal reference electrode (IRE)) are found to be stable for more than 60 minutes when examined with saturated calomel electrode. Optimal operating voltage for PB and CoPC screen printed sensors are determined by hydrodynamic voltammetric technique. Glucose oxidase is immobilized on the working electrodes by cross-linking method. PB mediated glucose sensor exhibits a sensitivity of 5.60 mA cm(-2)/mM for the range, 10 to 1000 mu M. Sensitivity of CoPC mediated glucose sensor is found to be 5.224 mu A cm(-2)/mM and amperometeric response is linear for the range, 100 to 1500 mu M. Interference studies on the fabricated glucose sensors are conducted with species like uric acid and ascorbic acid. PB mediated sensors showed a completely interference-free behavior. The sensing characteristics of PB mediated glucose sensors are also studied in diluted human serum samples and the results are compared with the values obtained through standard clinical method. The co-efficient of variation is found to be less than 5%. (C) 2015 The Electrochemical Society. All rights reserved.

Folding of Protein L with Implications for Collapse in the Denatured State Ensemble

A fundamental question in protein folding is whether the coil to globule collapse transition occurs during the initial stages of folding (burst phase) or simultaneously with the protein folding transition. Single molecule fluorescence resonance energy transfer (FRET) and small-angle X-ray scattering (SAXS) experiments disagree on whether Protein L collapse transition occurs during the burst phase of folding. We study Protein L folding using a coarse-grained model and molecular dynamics simulations. The collapse transition in Protein L is found to be concomitant with the folding transition. In the burst phase of folding, we find that FRET experiments overestimate radius of gyration, R-g, of the protein due to the application of Gaussian polymer chain end-to-end distribution to extract R-g from the FRET efficiency. FRET experiments estimate approximate to 6 angstrom decrease in R-g when the actual decrease is approximate to 3 angstrom on guanidinium chloride denaturant dilution from 7.5 to 1 M, thereby suggesting pronounced compaction in the protein dimensions in the burst phase. The approximate to 3 angstrom decrease is close to the statistical uncertainties of the R-g data measured from SAXS experiments, which suggest no compaction, leading to a disagreement with the FRET experiments. The transition-state ensemble (TSE) structures in Protein L folding are globular and extensive in agreement with the Psi-analysis experiments. The results support the hypothesis that the TSE of single domain proteins depends on protein topology and is not stabilized by local interactions alone.

Mitochondria-Targeting Iron(III) Catecholates for Photoactivated Anticancer Activity under Red Light

Iron(III) catecholates Fe(R-bpa)(R-dopa)Cl] (1, 2) with a triphenylphosphonium (TPP) moiety, where R-bpa is 2-(TPP-N,N-bis((pyridin-2-yl)methyl)ethanamine) chloride (TPPbpa) and R-dopa is 4-{2-(anthracen-9-yl)methylamino]ethyl}benzene-1,2-diol (andopa, 1) or 4-{2-(pyren-1-yl)-methylamino]ethyl}benzene-1,2-diol (pydopa, 2), were synthesized and their photocytotoxicity studied. Complexes 3 and 4 with phenyl-N,N-bis(pyridin-2-yl)methyl]methanamine (phbpa) were used as controls. The catecholate complexes showed an absorption band near 720 nm. The 5e(-) paramagnetic complexes showed a Fe-III/Fe-II irreversible response near -0.45 V and a quasi-reversible catechol/semiquinone couple near 0.5 V versus saturated calomel electrode (SCE) in DMF/0.1 M tetrabutylammonium perchlorate. They showed photocytotoxicity in red/visible light in HeLa, HaCaT, MCF-7, and A549 cells. Complexes 1 and 2 displayed mitochondrial localization, reactive oxygen species (ROS) generation under red light, and apoptotic cell death. Control complexes 3 and 4 exhibited uniform distribution throughout the cell. The complexes showed DNA photocleavage under red light (785 nm), forming hydroxyl radicals as the ROS.

Anaesthetic impairment of immune function is mediated via GABA(A) receptors.

BACKGROUND: GABA(A) receptors are members of the Cys-loop family of neurotransmitter receptors, proteins which are responsible for fast synaptic transmission, and are the site of action of wide range of drugs. Recent work has shown that Cys-loop receptors are present on immune cells, but their physiological roles and the effects of drugs that modify their function in the innate immune system are currently unclear. We are interested in how and why anaesthetics increase infections in intensive care patients; a serious problem as more than 50% of patients with severe sepsis will die. As many anaesthetics act via GABA(A) receptors, the aim of this study was to determine if these receptors are present on immune cells, and could play a role in immunocompromising patients. PRINCIPAL FINDINGS: We demonstrate, using RT-PCR, that monocytes express GABA(A) receptors constructed of α1, α4, β2, γ1 and/or δ subunits. Whole cell patch clamp electrophysiological studies show that GABA can activate these receptors, resulting in the opening of a chloride-selective channel; activation is inhibited by the GABA(A) receptor antagonists bicuculline and picrotoxin, but not enhanced by the positive modulator diazepam. The anaesthetic drugs propofol and thiopental, which can act via GABA(A) receptors, impaired monocyte function in classic immunological chemotaxis and phagocytosis assays, an effect reversed by bicuculline and picrotoxin. SIGNIFICANCE: Our results show that functional GABA(A) receptors are present on monocytes with properties similar to CNS GABA(A) receptors. The functional data provide a possible explanation as to why chronic propofol and thiopental administration can increase the risk of infection in critically ill patients: their action on GABA(A) receptors inhibits normal monocyte behaviour. The data also suggest a potential solution: monocyte GABA(A) receptors are insensitive to diazepam, thus the use of benzodiazepines as an alternative anesthetising agent may be advantageous where infection is a life threatening problem.

气相扩散法生长溶菌酶晶体的动态光散射研究

首次采用动态光散射研究了气相扩散法生长溶菌酶晶体 .实验中采用了两种溶解溶菌酶的方法，所得实验结果是有区别的 .这种区别表明了 NaCl对溶菌酶分子间相互作用产生十分重要的影响 .实验结果表明，晶体生长过程中，溶液中溶菌酶始终保持单分子与两分子聚集体的状态，这种状态是生长晶体的基础 .

Saline water intrusion from deep artesian sources in McGregor Isles area of Lee County, Florida

Upward leakage of saline water from an artesian aquifer below 1,500 feet has caused an increase in chloride concentration in the lower Hawthorn aquifer from less than 1,000 mg/1 (milligrams per liter) to values ranging from about 1,300 to 15,000 mg/1. Similarly the higher temperatures of the intruding water has caused an increase in water temperatures in the aquifer from 82"F to values ranging from 83 to 93"F. The intruding water moves upward either through the open bore hole of deep wells or test holes, or along a fault or fracture system, which has been identified in the area. From these points of entry into the lower Hawthorn aquifer, the saline water spreads laterally toward the south and southeast, but is generally confined to components of the fault system. The saline water moves upward from the lower Hawthorn aquifer into the upper Hawthorn aquifer through the open bore hole of wells, which connect the aquifers. This movement has resulted in an increase in chloride from less than 200 mg/1 in the unaffected parts of the upper Hawthorn aquifer to values commonly ranging from about 300 to more than 3,000 mg/1 in parts of the aquifer affected by upward leakage. The upper Hawthorn aquifer is the principal source of ground-water supply for public water-supply systems in western Lee County. Similar effects have been noted in the water-table aquifer, where chloride increased from less than 100 to concentrations ranging from about 500 to more than 5,000 mg/1. This was caused by the downward infiltration of water discharged at land surface from wells tapping the lower Hawthorn aquifer. The spread of saline water throughout most of the McGregor Isles area is continuing as of 1971. (40 page document)

Interim report on the progress of an inventory of artesian wells in Florida: leading to the enforcement of sections 370.051 - 370.54, Florida statues

One of the causes of lower artesian pressure, water waste and aquifer contamination is the misuse and insufficient care of artesian wells. In 1953, Senate Bill No. 57, entitled "An Act to Protect and Control the Artesian Waters of the State" (see Appendix) became a law. This law was passed through the efforts exerted by leading members of the Senate and the House of Representatives, who understood the need for a wise and controlled expenditure of our most valuable natural resource. The State Geologist and his authorized representatives were designated by this law to enforce this conservation measure; however, no financial provision was included for the 1953-55 biennium. The proposed program of the Florida Geological Survey for this biennium did not include the funds nor provide any full-time personnel for the enforcement of this statute. As a result, little actual work was accomplished during these two years, although much time was given to planning and discussion of the problem. Realizing that this program could provide additional basic data needed in the analysis of the water-supply problem, the State Geologist sought and was granted by the 1955 Legislature adequate funds with which to activate the first phase of the enforcement of Florida Statute No. 370.051-054. Enumerated below is a summary of the progress made on this investigation as outlined previously: 1. Data have been collected on 967 wildly flowing wells in 22 counties. 2. Chloride determinations have been run on 850 of the 967 wells. 3. Of the 967 wells, 554 have chlorides in excess of the 250 ppm, the upper limit assigned by the State Board of Health for public consumption. 4. Water escapes at the rate of 37, 762 gallons per minute from these 967 wells. This amounts to 54, 377, 280 gallons per day. The investigation is incomplete at this time; therefore, no final conclusions can be reached. However, from data already collected, the following recommendations are proposed: 1. That the present inventory of wildly flowing wells be completed for the entire State. 2. That the current inventory of wildly flowing wells be expanded at the conclusion of the present inventory to include all flowing wells. 3. That a complete statewide inventory program be established and conducted in cooperation with the Ground Water Branchof the U.S. Geological Survey. 4. That the enforcement functions as set down in Sections 370.051/.054, Florida Statutes, be separated from the program to collect water-resource data and that these functions be given to the Water Resources Department, if such is created (to be recommended by the Water Resources Study Commission in a water policy law presented to the 1957 Legislature). 5. That the research phase (well inventory) of the program remain under the direction of the Florida Geological Survey. (PDF contains 204 pages.)

Seed Longevity of Melaleuca quinquenervia: A Burial Experiment in South Florida

Burial and removal techniques with seed bags were used to examine the viability and longevity of Melaleuca quinquenervia seeds at four field sites representing different soil types and hydrological conditions in South Florida. Seed viability was determined over different burial durations in the soil through a combination of germination tests and 2,3,5-triphenyl- tetrazolium chloride (TTC) treatments. Control seeds kept dry at 25 C in the laboratory maintained same viability of ca. 15% over the 3-year study. In the field, seed viability decreased with increased burial duration.(PDF has 4 pages.)