Trace amounts of nickel in belowground rhizomes of Vaccinium myrtillus L. decrease anthocyanin concentrations in aerial shoots without water stress

Nickel (Ni) may impair plant water balance through detrimental effects on the belowground level. Bilberry (Vaccinium myrtillus L.) plants were grown in a mesic heath forest-type soil and subjected to Ni sulphate (NiSO₄·6H₂O) concentrations of 0, 10, 50, 100 and 500 mg m⁻² during an entire growing season in northern Finland (65°N). Biomass of belowground rhizomes, and tissue water content (TWC) and anthocyanin concentrations of aerial shoots were determined from mature plants in order to study rhizospheric Ni stress, and its possible long-distance effects on aerial shoots. As the major proportion of biomass of bilberry is invested in belowground parts, it was hypothesised that Ni-induced rhizospheric disturbance causes water stress in aerial shoots and increases their anthocyanin concentrations for osmotic regulation. Uptake of Ni from the soil to the rhizome and aerial shoots was measured with X-ray fluorescence spectrometry. Ni concentrations in the soil and rhizome exhibited a dose–response relationship, but the concentrations in the rhizome were about 10-fold lower (<3 mg Ni kg⁻¹) than those in the soil (<30 mg Ni kg⁻¹). Translocation of Ni from the rhizome to aerial shoots did not occur, as Ni concentrations in shoots remained at 1 mg Ni kg⁻¹. Although Ni concentrations in the rhizome were below the threshold values of Ni toxicity (i.e. 10–50 mg Ni kg⁻¹), Ni decreased the rhizome biomass. Anthocyanins decreased in aerial shoots along with the Ni accumulation in the rhizome, while TWC was unaffected. The result suggests that anthocyanins are not involved in osmotic regulation under Ni stress, since anthocyanins in aerial shoots responded to the Ni concentrations in the rhizome despite the lack of water stress.     

Isoelectric focusing reveals additional casein variants in German sheep breeds

Isoelectric focusing (IEF) was applied for screening milk protein variants in milk samples from altogether 1078 sheep of different breeds, in detail Black Faced Mutton sheep (SKF; n = 57), East Friesian Milk sheep (OMS; n = 254), Gray Horned Heath (GGH; n = 190), Merinoland sheep (MLS; n = 363), Merino Mutton sheep (MMS; n = 88), and Rhön sheep (RHO; n = 126). Besides the known genetic variants of α_s1-casein (CSN1S1) (A, C, D), α_s2-casein (CSN1S2) (A, B), and β-lactoglobulin (LGB) (A, B, C) additional variants could be demonstrated in CSN1S1 (H, I) and CSN1S2 (C, D) and their genetic control confirmed by segregation analyses. CSN1S1*H corresponds to a previously mentioned phenotype “X” occurring in OMS, whereas CSN1S1*I was identified for the first time in GGH. CSN1S2*C appeared in OMS, GGH, MLS, and RHO in low frequencies and CSN1S2*D in MLS. Within LGB all three alleles occurred in Merino breeds while α-lactalbumin (LAA) and κ-CN (CSN3) were monomorph at protein level. The haplotype CSN1S1*C–CSN1S2*A was predominant in five out of six breeds with frequencies between 0.325 and 0.919.     

Evaluation of the association of mercury(II) with some dicysteinyl tripeptides

The present study was undertaken to gain insight into the associations of mercury(II) with dicysteinyl tripeptides in buffered media at pH 7.4. We investigated the effects of increasing the distance between cysteinyl residues on mercury(II) associations and complex formations. The peptide–mercury(II) formation constants and their associated thermodynamic parameters in 3-(N-morpholino)propanesulfonic acid (MOPS) buffered solutions were evaluated by isothermal titration calorimetry. Complexes formed in different relative ratios of mercury(II) to cysteinyl peptides in ammonium formate buffered solutions were characterized by LTQ Orbitrap mass spectrometry. The results from these studies show that n-alkyl dicysteinyl peptides (CP 1–4), and an aryl dicysteinyl peptide (CP 5) can serve as effective “double anchors” to accommodate the coordination sites of mercury(II) to form predominantly one-to-one Hg(peptide) complexes. The aryl dicysteinyl peptide (CP 5) also forms the two-to-two Hg₂(peptide)₂ complex. In the presence of excess peptide, Hg(peptide)₂ complexes are also detected. Notably, increasing the distance between the ligating groups or “anchor points” in CP 1–5 does not significantly affect their affinity for mercury(II). However, the enthalpy change (ΔH) values (ΔH₁ ∼ −91 kJ mol⁻¹ and ΔH₂ ∼ −66 kJ mol⁻¹) for complex formation between CP 4 and 5 with mercury(II) are about one and a half times larger than the related values for CP 1, 2 and 3 (ΔH₁ ∼ −66 kJ mol⁻¹ and ΔH₂ ∼ 46 kJ mol⁻¹). The corresponding entropy change (ΔS) values (ΔS₁ ∼ −129 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −116 J K⁻¹ mol⁻¹) of the structurally larger dicysteinyl peptides CP 4 and 5 are less entropically favorable than for CP 1, 2 and 3 (ΔS₁ ∼ −48 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −44 J K⁻¹ mol⁻¹). Generally, these associations result in a decrease in entropy, indicating that these peptide–mercury complexes potentially form highly ordered structures. The results from this study show that dicysteinyl tripeptides are effective in binding mercury(II) and they are promising motifs for the design of multi-cysteinyl peptides for binding more than one mercury(II) ion per peptide.     

Origin of magnetization tunneling in single-molecule magnets as determined by single-crystal high-frequency EPR

We review an extensive body of single-crystal high-frequency electron paramagnetic resonance (HFEPR) data in order to determine the transverse spin Hamiltonian parameters that control the tunneling of the direction of magnetization in a variety of integer and half-integer-spin single-molecule magnets (SMMs). The SMMs studied are members of the following families: S = 9/2 [Mn₄O₃Cl]⁶⁺; S = 5 [Mn₃NiO₄]⁶⁺; S = 6 [Mn₃ZnO₄]⁶⁺; and S = 4 [Ni₄(OR)₄]⁴⁺. HFEPR spectra for the half-integer S = 9/2 Mn₄ complexes that have C₃ symmetry do not provide measurable evidence for transverse spin Hamiltonian terms. This finding is consistent with the relatively large coercive field seen in the magnetization hysteresis loops for these complexes. On the other hand, a low symmetry S = 9/2 complex exhibits a much faster rate of ground-state magnetization tunneling, in agreement with HFEPR spectra for a powder sample that gives a rhombic zero-field splitting (ZFS) parameter of E = 0.140 cm^?1. The S = 5 Mn₃Ni systems exhibit magnetization tunneling that is much faster than seen for the high-symmetry S = 9/2 Mn₄ complexes. This can be attributed to their integer-spin ground states. Like the C₃ symmetry Mn₄ SMMs, the HFEPR spectra for high-symmetry Mn₃Ni complexes do not provide measurable evidence for transverse ZFS terms. However, the spectra exhibit broad peaks, suggesting distributions in the local molecular environments brought about by disordered solvate molecules. This disorder likely explains the fast tunneling in the high-symmetry S = 5 Mn₃Ni systems, though one cannot rule out fourth- (and higher-) order interactions that cannot be detected by HFEPR due to the broad resonances. The one S = 6 Mn₃Zn complex shows an even faster rate of tunneling compared to the isostructural S = 5 Mn₃Ni complex. Finally, the S = 4 [Ni(hmp)(dmb)Cl]₄ complex provides unique insights into the origin of fourth- (and higher-) order interactions found for many SMMs on the basis of analysis using a giant spin Hamiltonian (GSH) approximation. We conclude that the fourth-order anisotropy found for the S = 4 ground state of [Ni(hmp)(dmb)Cl]₄ originates from the second-order ZFS interactions associated with the individual Ni^II ions, but only as a result of higher-order processes that occur via S-mixing between the ground state and higher-lying (S < 4) spin-multiplets. The S-mixing is relatively strong in this system because of comparable exchange and anisotropy energy scales. The relatively fast tunneling is a direct consequence of this S-mixing, as opposed to any intrinsic fourth-order (spin–orbit) anisotropy associated with Ni^II.     

Synthesis,structures and characterization of the dinuclear nickel(II) complexes containing N,N,N′,N′-tetrakis[(1-ethyl-2-benzimidazolyl)methyl]-2-hydroxy-1,3-diaminopropane and an azide ion

Two dinuclear nickel(II) complexes, [Ni₂(L-Et)(N₃)(H₂O)₃](NO₃)₂ · 2H₂O (1) and [Ni₂(L-Et)(μ-1,3-N₃)(H₂O)₂](NO₃)₂ · 4H₂O (2) containing (HL-Et = N,N,N′,N′-tetrakis[(1-ethyl-2-benzimidazolyl)methyl]-2-hydroxy-1,3-diaminopropane), have been synthesized and characterized by their IR and UV–Vis spectra and magnetic susceptibilities. The crystal structures of [Ni₂(L-Et)(N₃)(H₂O)₃](NO₃)₂ · CH₃OH (1′) and [Ni₂(L-Et)(μ-1,3-N₃)(H₂O)₂](NO₃)₂ · 2C₂H₅OH (2′) similar to 1 and 2 were determined by X-ray crystallography. In 1′, the two nickel(II) ions are bridged by only an alkoxo group of L-Et⁻, while an azido and an alkoxo connect two nickel(II) ions in 2′. Magnetic susceptibility measurements (2–300 K) showed a weak ferromagnetic exchange coupling between the two nickel(II) ions (2J = 10.1 cm⁻¹) for 1. On the other hand, antiferromagnetic interactions were observed for 2 (2J = −15.8 cm⁻¹).     

Synthesis and in vivo evaluation of [18F]2-(4-(4-(2-(2-fluoroethoxy)phenyl)piperazin-1-yl)butyl)-4-methyl-1,2,4-triazine-3,5(2H,4H)-dione ([18F]FECUMI-101) as an imaging probe for 5-HT1A receptor agonist in nonhuman primates

The 5-HT_1AR partial agonist PET radiotracer, [¹¹C]CUMI-101, has advantages over an antagonist radiotracer as it binds preferentially to the high affinity state of the receptor and thereby provides more functionally meaningful information. The major drawback of C-11 tracers is the lack of cyclotron facility in many health care centers thereby limiting widespread clinical or research use. We identified the fluoroethyl derivative, 2-(4-(4-(2-(2-fluoroethoxy)phenyl)piperazin-1-yl)butyl)-4-methyl-1,2,4-triazine-3,5(2H,4H)dione (FECUMI-101) (K_i = 0.1 nM; E_max = 77%; EC₅₀ = 0.65 nM) as a partial agonist 5-HT_1AR ligand of the parent ligand CUMI-101. FECUMI-101 is radiolabeled with F-18 by O-fluoroethylation of the corresponding desmethyl analogue (1) with [¹⁸F]fluoroethyltosylate in DMSO in the presence of 1.6 equiv of K₂CO₃ in 45 ± 5% yield (EOS). PET shows [¹⁸F]FECUMI-101 binds specifically to 5-HT_1AR enriched brain regions of baboon. The specificity of [¹⁸F]FECUMI-101 binding to 5-HT_1AR was confirmed by challenge studies with the known 5-HT_1AR ligand WAY100635. These findings indicate that [¹⁸F]FECUMI-101 can be a viable agonist ligand for the in vivo quantification of high affinity 5-HT_1AR with PET.     

Structure and magnetic properties of a trinuclear nickel(II) complex with benzenetricarboxylate bridge

Novel trinuclear Ni(II) complex [Ni₃(pmdien)₃(btc)(H₂O)₃](ClO₄)₃ · 4H₂O, 1 where pmdien = N,N,N′,N′,N″-pentamethyldiethylenetriamine, H₃btc = 1,3,5-benzenetricarboxylic (trimesic) acid, has been prepared and structurally characterized. Three nickel atoms are bridged by btc trianion and their coordination sphere is completed by three N atoms of pmdien and O atom of the water molecule. The three nickel(II) magnetic centers are equivalent and their coordination spheres are completed to deformed octahedrons. Magnetic susceptibility was measured over the temperature range 1.8–300 K and zJ′ = ?0.19 cm^?1, D = 3.79 cm^?1, g = 2.18 parameters were calculated.     

Purification,kinetic and thermodynamic characterization of soluble acid invertase from sugarcane (Saccharum officinarum L.)

We report for the first time kinetic and thermodynamic properties of soluble acid invertase (SAI) of sugarcane (Saccharum officinarum L.) salt sensitive local cultivar CP 77-400 (CP-77). The SAI was purified to apparent homogeneity on FPLC system. The crude enzyme was about 13 fold purified and recovery of SAI was 35%. The invertase was monomeric in nature and its native molecular mass on gel filtration and subunit mass on SDS-PAGE was 28 kDa. SAI was highly acidic having an optimum pH lower than 2. The acidic limb was missing. Proton transfer (donation and receiving) during catalysis was controlled by the basic limb having a pKa of 2.4. Carboxyl groups were involved in proton transfer during catalysis. The kinetic constants for sucrose hydrolysis by SAI were determined to be: k_m = 55 mg ml^?1, k_cat = 21 s^?1, k_cat/k_m = 0.38, while the thermodynamic parameters were: ΔH* = 52.6 kJ mol^?1, ΔG* = 71.2 kJ mol^?1, ΔS* = ?57 J mol^?1 K^?1, ΔG*_E–S = 10.8 kJ mol^?1 and ΔG*_E–T = 2.6 kJ mol^?1. The kinetics and thermodynamics of irreversible thermal denaturation at various temperatures 53–63 °C were also determined. The half -life of SAI at 53 and 63 °C was 112 and 10 min, respectively. At 55 °C, surprisingly the half -life increased to twice that at 53 °C. ΔG*, ΔH* and ΔS* of irreversible thermal stability of SAI at 55 °C were 107.7 kJ mol^?1, 276.04 kJ mol^?1 and 513 J mol^?1K^?1, respectively.     

Hexavalent chromium removal from aqueous solution by adsorption on treated sawdust

The studies on adsorption of hexavalent chromium were conducted by varying various parameters such as contact time, pH, amount of adsorbent, concentration of adsorbate and temperature. The kinetics of adsorption of Cr(VI) ion followed pseudo second order. Langmuir adsorption isotherm was employed in order to evaluate the optimum adsorption capacity of the adsorbent. The adsorption capacity was found to be pH dependant. Sawdust was found to be very effective and reached equilibrium in 3 h (adsorbate concentration 30 mg l⁻¹). The rate constant has been calculated at 303, 308, 313 and 318 K and the activation energy (E_a) was calculated using the Arrhenius equation. Thermodynamic parameters such as standard Gibbs energy (ΔG°) and heat of adsorption (ΔH_r) were calculated. The ΔG° and ΔH_r values for Cr(VI) adsorption on the sawdust showed the process to be exothermic in nature. The percentage of adsorption increased with decrease in pH and showed maximum removal of Cr(VI) in the pH range 4.5–6.5 for an initial concentration of 5 mg l⁻¹.     

8-Substituted 1,3-dimethyltetrahydropyrazino[2,1-f]purinediones: Water-soluble adenosine receptor antagonists and monoamine oxidase B inhibitors

Multitarget approaches, i.e., addressing two or more targets simultaneously with a therapeutic agent, are hypothesized to offer additive therapeutic benefit for the treatment of neurodegenerative diseases. Validated targets for the treatment of Parkinson’s disease are, among others, the A_2A adenosine receptor (AR) and the enzyme monoamine oxidase B (MAO-B). Additional blockade of brain A₁ ARs may also be beneficial. We recently described 8-benzyl-substituted tetrahydropyrazino[2,1-f]purinediones as a new lead structure for the development of such multi-target drugs. We have now designed a new series of tetrahydropyrazino[2,1-f]purinediones to extensively explore their structure–activity-relationships. Several compounds blocked human and rat A₁ and A_2AARs at similar concentrations representing dual A₁/A_2A antagonists with high selectivity versus the other AR subtypes. Among the best dual A₁/A_2AAR antagonists were 8-(3-(4-chlorophenyl)propyl)-1,3-dimethyl-6,7,8,9-tetrahydropyrazino[2,1-f]purine-2,4(1H,3H)-dione (41, K_i human A₁: 65.5 nM, A_2A: 230 nM; K_i rat A₁: 352 nM, A_2A: 316 nM) and 1,3-dimethyl-8-((2-(thiophen-2-yl)thiazol-4-yl)methyl)-6,7,8,9-tetrahydropyrazino[2,1-f]purine-2,4(1H,3H)-dione (57, K_i human A₁: 642 nM, A_2A: 203 nM; K_i rat A₁: 166 nM, A_2A: 121 nM). Compound 57 was found to be well water-soluble (0.7 mg/mL) at a physiological pH value of 7.4. One of the new compounds showed triple-target inhibition: (R)-1,3-dimethyl-8-(2,1,3,4-tetrahydronaphthalen-1-yl)-6,7,8,9-tetrahydropyrazino[2,1-f]purine-2,4(1H,3H)-dione (49) was about equipotent at A₁ and A_2AARs and at MAO-B (K_i human A₁: 393 nM, human A_2A: 595 nM, IC₅₀ human MAO-B: 210 nM) thus allowing future in vivo explorations of the intended multi-target approach.     

The dinoflagellate Akashiwo sanguinea will benefit from future climate change: The interactive effects of ocean acidification,warming and high irradiance on photophysiology and hemolytic activity

Due to global climate change, marine phytoplankton will likely experience low pH (ocean acidification), high temperatures and high irradiance in the future. Here, this work report the results of a batch culture experiment conducted to study the interactive effects of elevated CO₂, increased temperature and high irradiance on the harmful dinoflagellate Akashiwo sanguinea, isolated at Dongtou Island, Eastern China Sea. The A. sanguinea cells were acclimated in high CO₂ condition for about three months before testing the responses of cells to a full factorial matrix experimentation during a 7-day period. This study measured the variation in physiological parameters and hemolytic activity in 8 treatments, representing full factorial combinations of 2 levels each of exposure to CO₂ (400 and 1000 μatm), temperature (20 and 28 °C) and irradiance (50 and 200 μmol photons m⁻² s⁻¹). Sustained growth of A. sanguinea occurred in all treatments, but high CO₂ (HC) stimulated faster growth than low CO₂ (LC). The pigments (chlorophyll a and carotenoid) decreased in all HC treatments. The quantum yield (F_v/F_m) declined slightly in all high-temperature (HT) treatments. High irradiance (HL) induced the accumulation of ultraviolet-absorbing compounds (UV_abc) irrespective of temperature and CO₂. The hemolytic activity in the LC treatments, however, declined when exposed to HT and HL, but HC alleviated the adverse effects of HT and HL on hemolytic activity. All HC and HL conditions and the combinations of high temperature*high light (HTHL) and high CO₂*high temperature*high light (HCHTHL) positively affected the growth in comparison to the low CO₂*low temperature*low light (LCLTLL) treatment. High temperature (HT), high light (HL) and a combination of HT*HL, however, negatively impacted hemolytic activity. CO₂ was the main factor that affected the growth and hemolytic activity. There were no significant interactive effects of CO₂*temperature*irradiance on growth, pigment, F_v/F_m or hemolytic activity, but there were effects on P_m, α, and E_k. If these results are extrapolated to the natural environment, it can be hypothesized that A. sanguinea cells will benefit from the combination of ocean acidification, warming, and high irradiance that are likely to occur under future climate change. It is assumed that faster growth and higher hemolytic activity and UV_abc of this species will occur under future conditions compared with those the current CO₂ (400 μatm) and temperature (20 °C) conditions.     

Synthesis,X-ray structures,and magnetic properties of heterometal dinuclear Cr(III)–M(II) complexes

New 3,5-bis(2-pyridyl)pyrazolato (bpypz) bridged heterometal dinuclear complexes [(nta)Cr(μ-bpypz)M^II(picen)]⁺ (M = Mn(II), Ni(II)) and [(acac)₂Cr(μ-bpypz)Ni^II(picen)]²⁺ (nta = nitrilotriacetate, picen = N,N′-bis(2-pyridylmethyl)ethylenediamine, acac = acetylacetate) were synthesized and characterized by the X-ray analysis, ESI-MS and the magnetic measurements, and/or ²H NMR spectra. The molecular structures were compared from a viewpoint of the conformation of the picen depending on M^II ionic radii or different modes of hydrogen bonds. The picen in [(nta)Cr(μ-bpypz)Mn^II(picen)]BF₄ takes an abnormal conformation with intramolecular bifurcated three-center hydrogen bonds between two carboxylate oxygens of nta and an amine proton of the picen as found for the previously reported corresponding Fe(II) complex [K. Ni-iya, A. Fuyuhiro, T. Yagi, S. Nasu, K. Kuzushita, S. Morimoto, S. Kaizaki, Bull. Chem. Soc. Jpn. 74 (2001) 1891]. On the other hand, for both Ni(II)-nta and Ni(II)-acac complexes, the picen takes a normal conformation with only a two-center hydrogen bond between non-bridging ligands. The magneto-structural relation is discussed for the Cr(III)–Ni(II) complexes in connection with the orthogonality or orbital overlap arising from the difference in distortion around Cr(III) moiety.     

Synthesis,structures and magnetic properties of pentanuclear and trinuclear heterometallic complexes with bended and linear cyano-bridges (tren = tris(2-aminoethyl)amine)

The reaction of [Cu(tren)(OH₂)](ClO₄)₂ with KCN gave a mononuclear complex [Cu(tren)(CN)](ClO₄) (1) (tren = tris(2-aminoethyl)amine). Using 1 as a building block, one pentanuclear compound, [{Cu(tren)(NC)}₄Ni](ClO₄)₆ (2) and two trinuclear complexes, [{Cu(tren)NC}₂Co(tren)](ClO₄)₅ · 2H₂O (3), [{Cu(tren)CN}₂NiL](ClO₄)₄ (4) (L = 3,10-bis(2-hydroxyethyl)-1,3,5,8,10,12-hexaazacyclotetradecane) were prepared and characterized by single crystal X-ray analysis. In 1, Cu(II) atom adopts a distorted trigonal bipyramidal (TBP) geometry. In 2, the Ni(II) atom occupies the center of the pentanuclear compound with a square-planar coordination geometry. In 3, the six-coordinated Co(III) atom presents a distorted octahedral geometry with four nitrogen atoms from tren and two carbon atoms of bridged cyano groups in cis-positions. In 4, the nickel atom is located in an inversion center and coordinated with two [(tren)CuCN]⁺ moieties through cyano-bridging ligands. Magnetic susceptibility measurements of 2–4 show that the magnetic interactions between the heterometallic ions are antiferromagnetical coupling through the cyano bridges with g = 2.25, J = −0.142 cm⁻¹ and J′ = −0.167 cm⁻¹ for 2, g = 2.06, J = −0.094 cm⁻¹ for 3, and g = 2.20, J = −33.133 cm⁻¹ for 4. The correlations between the structures and the J values are discussed.     

Color variation of rubberwood clones and cypress infected by Gloeophyllum striatum and Phanerochaete chrysosporium

The objective of this work was to examine quantitatively the color changes of six clones of Hevea brasiliensis (IAN 717, IAN 873, GT 711, AVROS 1301, RRIM 600, and Tjir 16) and two cypresses (Cupressus sp. and Cupressus glauca) upon fungal attack. Samples were exposed to the fungi Gloeophyllum striatum and Phanerochaete chrysosporium according to ASTM D 2017. Wood color was measured in the L¹a¹b system. Changes in the colorimetric parameters were calculated by the difference between attacked and control samples. None of the rubberwood clones showed weight loss below 25% and therefore could not be classified as resistant, but clones AVROS 1301, RRIM 600, and Tjir 16 were borderline. Both cypresses were resistant to P. chrysosporium, but only Cupressus sp. was resistant to G. striatum. The red tint of all samples exposed to both fungi was slightly intensified (Δa¹ > 0), except for RRIM 600. The yellowish tint was intensified as well, except for RRIM 600 and Tjir 16 exposed to G. striatum. Samples attacked by G. striatum were darker than controls (ΔL¹ < 0), while samples infected by P. chrysosporium were lighter (ΔL¹ > 0). Color variation mean values (ΔE¹) ranged from 3.98 to 12.33, attaining the levels appreciable to very appreciable color difference. Extraction of the attacked samples with ethanol–toluene plus hot water removed red and yellow chromophore pigments, causing color variations perceptible to the naked eye.     

Pyrrolidine analogs of GZ-793A: Synthesis and evaluation as inhibitors of the vesicular monoamine transporter-2 (VMAT2)

Central heterocyclic ring size reduction from piperidinyl to pyrrolidinyl in the vesicular monoamine transporter-2 (VMAT2) inhibitor GZ-793A and its analogs resulted in novel N-propane-1,2(R)-diol analogs 11a–i. These compounds were evaluated for their affinity for the dihydrotetrabenazine (DTBZ) binding site on VMAT2 and for their ability to inhibit vesicular dopamine (DA) uptake. The 4-difluoromethoxyphenethyl analog 11f was the most potent inhibitor of [³H]-DTBZ binding (K_i = 560 nM), with 15-fold greater affinity for this site than GZ-793A (K_i = 8.29 μM). Analog 11f also showed similar potency of inhibition of [³H]-DA uptake into vesicles (K_i = 45 nM) compared to that for GZ-793A (K_i = 29 nM). Thus, 11f represents a new water-soluble inhibitor of VMAT function.     

Design and synthesis of N6-substituted-4′-thioadenosine-5′-uronamides as potent and selective human A3 adenosine receptor agonists

On the basis of a bioisosteric rationale, 4′-thionucleoside analogues of IB-MECA (N⁶-(3-Iodo-benzyl)-9-(5′-methylaminocarbonyl-β-d-ribofuranosyl)adenine), which is a potent and selective A₃ adenosine receptor (AR) agonist, were synthesized from d-gulonic acid γ-lactone. The 4′-thio analogue (5h) of IB-MECA showed extremely high binding affinity (K_i = 0.25 nM) at the human A₃AR and was more potent than IB-MECA (K_i = 1.4 nM). Bulky substituents at the 5′-uronamide position, such as cyclohexyl and 2-methylbenzyl, in this series of 2-H nucleoside derivatives were tolerated in A₃AR binding, although small alkyl analogues were more potent.     

Single-step purification of recombinant green fluorescent protein on expanded beds of immobilized metal affinity chromatography media

Immobilized metal ion affinity chromatography (IMAC) in expanded bed mode is used for purifying recombinant green fluorescent protein (GFP) overexpressed in Escherichia coli. The purification is carried out on two different matrices, i.e. Ni²⁺ Streamline™ and Ni²⁺ cross-linked alginate beads. The binding isotherms to both IMAC media followed the Langmuir model. The maximum binding capacity (q_max) of Ni²⁺ Streamline™ and Ni²⁺ cross-linked alginate for the GFP was 1,42,860 FU ml⁻¹ and 18,000 FU ml⁻¹, respectively. The expanded bed column chromatography using Ni²⁺ Streamline™ gave 2.7-fold purification with 89% of GFP recovery, while Ni²⁺ alginate gave 3.1-fold purification with 91% of GFP recovery. SDS-PAGE of purified GFP in both cases showed single band. The results obtained in the expanded bed chromatography are compared with those obtained in packed bed chromatography.     

Noninvasive prediction of vertebral body compressive strength using nonlinear finite element method and an image based technique

Noninvasive prediction of vertebral body strength under compressive loading condition is a valuable tool for the assessment of clinical fractures. This paper presents an effective specimen-specific approach for noninvasive prediction of human vertebral strength using a nonlinear finite element (FE) model and an image based parameter based on the quantitative computed tomography (QCT). Nine thoracolumbar vertebrae excised from three cadavers with an average age of 42 years old were used as the samples. The samples were scanned using the QCT. Then, a segmentation technique was performed on each QCT sectional image. The segmented images were then converted into three-dimensional FE models for linear and nonlinear analyses. A new material model was implemented in our nonlinear model being more compatible with real mechanical behavior of trabecular bone. A new image based MOS (Mechanic of Solids) parameter named minimum sectional strength ((σ_uA)_min) was used for the ultimate compressive strength prediction. Subsequently, the samples were destructively tested under uniaxial compression and their experimental ultimate compressive strengths were obtained. Results indicated that our new implemented FE model can predict ultimate compressive strength of human vertebra with a correlation coefficient (R² = 0.94) better than usual linear and nonlinear FE models (R² = 0.83 and 0.85 respectively). The image based parameter introduced in this study ((σ_uA)_min) was also correlated well with the experimental results (R² = 0.86). Although nonlinear FE method with new implemented material model predicts compressive strength better than the (σ_uA)_min, this parameter is clinically more feasible due to its simplicity and lower computational costs. This can make future applications of the (σ_uA)_min more justified for human vertebral body compressive strength prediction.     

Foliage type specific susceptibility to ozone in Picea abies,Pinus cembra and Larix decidua at treeline: A synthesis

Cumulative ozone uptake (COU, mmol m⁻²) and O₃ flux (FO₃, nmol m⁻² s⁻¹) were related to physiological, morphological and biochemical characteristics of field-grown mature evergreen Norway spruce [Picea abies (L.) Karst.], Cembran pine [Pinus cembra L.], and deciduous European larch [Larix decidua Mill.] trees at treeline. The threshold COU causing a statistically significant decline in photosynthetic capacity (A_max) ranged between 19.6 mmol m⁻² in current-year needles of evergreen conifers and 22.0 6 mmol m⁻² in short-shoot needles of deciduous L. decidua subjected to exposure periods of ≥84 and ≥43 days, respectively. The higher O₃ sensitivity of deciduous L. decidua than of evergreen P abies and P. cembra was associated with differences in FO₃ and specific leaf area (SLA), both being significantly higher in L. decidua. FO₃ was 5.9 nmol m⁻² s⁻¹ in L. decidua and 2.7 nmol m⁻² s⁻¹ in evergreen conifers. Species-dependent differences were also related to detoxification capacity expressed through total surface area based concentrations of reduced ascorbate and α-tocopherol that both increased with SLA. Findings suggest that differences in O₃ sensitivity between evergreen and deciduous conifers can be attributed to foliage type specific differences in SLA, the latter determining physiological and biochemical characteristics of the treeline conifers.