Absence of parallelism between polyamine and nucleic acid contents during induced growth of cucumber cotyledons

The cytokinins (benzyladenine or benzyladenosine) decreased spermidine and spermine contents despite increasing putrescine content, when administered to isolated cotyledons of Cucumis sativus L. var. Guntur in organ culture. KCl decreased putrescine contents, although marginally increasing polyamine contents. The cytokinins and/or KCl augmented nucleic acid biosynthesis and accumulation, resulting in enhanced growth and differentiation of the isolated cotyledons. These observations show that polyamine accumulation and growth are not always coupled.

Long-term outcome and extraintestinal manifestations in congenital chloride diarrhea

The rare autosomal recessive disease congenital chloride diarrhea (CLD) is caused by mutations in the solute carrier family 26 member 3 (SLC26A3) gene on chromosome 7q22.3-31.1. SLC26A3 encodes for an apical epithelial chloride-bicarbonate exchanger, the intestinal loss of which leads to profuse chloride-rich diarrhea, and a tendency to hypochloremic and hypokalemic metabolic alkalosis. Although untreated CLD is usually lethal in early infancy, the development of salt substitution therapy with NaCl and KCl in the late 1960s made the disease treatable. While the salt substitution allows normal childhood growth and development in CLD, data on long-term outcome have remained unclarified. One of the world s highest incidences of CLD 1:30 000 to 1:40 000 occurs in Finland, and CLD is part of the Finnish disease heritage. We utilized a unique sample of Finnish patients to characterize the long-term outcome of CLD. Another purpose of this study was to search for novel manifestations of CLD based on the extraintestinal expression of the SLC26A3 gene. This study on a sample of 36 patients (ages 10-38) shows that the long-term outcome of treated CLD is favorable. In untreated or poorly treated cases, however, chronic contraction and metabolic imbalance may lead to renal injury and even to renal transplantation. Our results demonstrate a low-level expression of SLC26A3 in the human kidney. Although SLC26A3 may play a minor role in homeostasis, post-transplant recurrence of renal changes shows the unlikelihood of direct transporter modulation in the pathogenesis of CLD-related renal injury. Options to resolve the diarrheal symptoms of CLD have been limited. Unfortunately, our pilot trial indicated the inefficacy of oral butyrate as well. This study reveals novel manifestations of CLD. These include an increased risk for hyperuricemia, inguinal hernias, and probably for intestinal inflammation. The most notable finding of this study is CLD-associated male subfertility. This involves a low concentration of poorly motile spermatozoa with abnormal morphology, high seminal plasma chloride with a low pH, and a tendency to form spermatoceles. That SLC26A3 immunoexpression appeared at multiple sites of the male reproductive tract in part together with the main interacting proteins cystic fibrosis transmembrane conductance regulator (CFTR) and sodium-hydrogen exchanger 3 (NHE3) suggests novel sites for the cooperation of these proteins. As evidence of the cooperation, defects occurring in any of these transporters are associated with reduced male fertility. Together with a finding of high sweat chloride in CLD, this study provides novel data on extraintestinal actions of the SLC26A3 gene both in the male reproductive tract and in the sweat gland. These results provide the basis for future studies regarding the role of SLC26A3 in different tissues, especially in the male reproductive tract. Fortunately, normal spermatogenesis in CLD is likely to make artificial reproductive technologies to treat infertility and even make unassisted reproduction possible.

Photocytotoxic Oxovanadium(IV) Complexes Showing Light-Induced DNA and Protein Cleavage Activity

Oxovanadium(IV) complexes [VO(L)(B)]Cl-2 (1-3), where L is bis(2-benzimidazolylmethyl)amine and B is 1,10-phenanthroline(phen),dipyrido[3,2-d:2',3'-f]quinoxaline(dpq) or dipyrido[3,2-a:2',3'-c]phenazine (dppz), have been prepared, characterized, and their photo-induced DNA and protein cleavage activity studied. The photocytotoxicity of complex 3 has been studied using adenocarcinoma A549 cells, The phen complex 1, structurally characterized by single-crystal X-ray crystallography, shows the presence of a vanadyl group in six-coordinate VON5 coordination geometry. The ligands L and phen display tridentate and bidentate N-donor chelating binding modes, respectively. The complexes exhibit a d-d band near 740 nm in 15% DMF-Tris-HCl buffer (pH 7.2). The phen and dpq complexes display an irreversible cathodic cyclic voltammetric response near -0.8 V in 20% DMF-Tris-HCl buffer having 0.1 M KCl as supporting electrolyte. The dppz complex 3 exhibits a quasi-reversible voltammogram near -0.6 V (vs SCE) that is assignable to the V(IV)-V(III)couple. The complexes bind to calf thymus DNA giving binding constant values in the range of 6.6 x 10(4)-2.9 x 10(5) M-1. The binding site size, thermal melting and viscosity binding data suggest DNA surface and/or groove binding nature of the complexes. The complexes show poor ``chemical nuclease'' activity in dark in the presence of 3-mercaptopropionic acid or hydrogen peroxide. The dpq and dppz complexes are efficient photocleavers of plasmid DNA in UV-A light of 365 nm via a mechanistic pathway that involves formation of both singlet oxygen and hydroxyl radicals. The complexes show significant photocleavage of DNA in near-IR light (>750 nm) via hydroxyl radical pathway. Among the three complexes, the dppz complex 3 shows significant BSA and lysozyme protein cleavage activity in UV-A light of 365 nm via hydroxyl radical pathway. The dppz complex 3 also exhibits photocytotoxicity in non-small cell lung carcinoma/human lung adenocarcinoma A549 cells giving IC50 value of 17 mu M in visible light(IC50 = 175 mu M in dark).

Oxovanadium(IV) complexes of phenanthroline bases: the dipyridophenazine complex as a near-IR photocytotoxic agent

Oxovanadium(IV) complexes [VOCl(B)(2)]Cl (1-3) of phenanthroline bases (B), viz. 1,10-phenanthroline (phen in 1), dipyrido[3,2-d: 2', 3'-f] quinoxaline (dpq in 2) and dipyrido[3,2-a: 2', 3'-c] phenazine (dppz in 3), have been prepared, characterized and their DNA and protein binding, photo-induced DNA and protein cleavage activity andm photocytotoxicity have been studied. Complex 2, structurally characterized by X-ray crystallography, shows the presence of a vanadyl group in VOClN4 coordination geometry. The dpq ligand displays a chelating mode of binding with a N-donor site trans to the oxo-group. The chloride ligand is cis to the oxo-group. The one-electron paramagnetic complexes show a d-d band near 715 nm in 15% DMF-Tris-HCl buffer. The complexes are redox active exhibiting a V(IV)/V(III) redox couple within -0.5 to -0.7 V vs. SCE in 20% DMF-Tris-HCl/0.1 M KCl. The complexes bind to calf thymus (CT) DNA in the order: 3 (dppz) > 2 (dpq) > 1 (phen). The binding data reveal the groove and/or partial intercalative DNA binding nature of the complexes. The complexes show chemical nuclease'' activity in the dark in the presence of 3-mercaptopropionic acid or hydrogen peroxide via a hydroxyl radical pathway. The dpq and dppz complexes are efficient photocleavers of DNA in UV-A light of 365 nm forming reactive singlet oxygen (O-1(2)) and hydroxyl radical ((OH)-O-center dot) species. Complexes 2 and 3 also show DNA cleavage activity in red light (> 750 nm) by an exclusive (OH)-O-center dot pathway. The complexes display a binding propensity to bovine serum albumin (BSA) protein giving K-BSA values in the range of 7.1 x 10(4)-1.8 x 10(5) M-1. The dppz complex 3 shows BSA and lysozyme protein cleavage activity in UV-A light of 365 nm via (OH)-O-center dot pathway. The dppz complex 3 exhibits significant PDT effect in human cervical cancer HeLa cells giving IC50 values of 1.0 mu M and 12.0 mu M in UV-A and visible light, respectively (IC50 = > 100 mu M in the dark).

Iron(III) Schiff base complexes of arginine and lysine as netropsin mimics showing AT-selective DNA binding and photonuclease activity

Iron(III) complexes [Fe(L)(2)]Cl (1-3), where L is monoanionic N-salicylidene-arginine (sal-argH for 1), hydroxynaphthylidene-arginine (nap-argH for 2) and N-salicylidene-lysine (sal-lysH for 3), were prepared and their DNA binding and photo-induced DNA cleavage activity studied. Complex 3 as its hexafluorophosphate salt [Fe(sal-lysH)(2)](PF6)center dot 6H(2)O (3a) was structurally characterized by single crystal Xray crystallography. The crystals belonged to the triclinic space group P-1. The complex has two tridentate ligands in FeN2O4 coordination geometry with two pendant cationic amine moieties. Complexes 1 and 2 with two pendant cationic guanidinium moieties are the structural models for the antitumor antibiotics netropsin. The complexes are stable and soluble in water. They showed quasi-reversible Fe(III)/Fe(II) redox couple near 0.6 V in H2O-0.1 M KCl. The high-spin 3d(5)-iron(III) complexes with mu(eff) value of similar to 5.9 mu(B) displayed ligand-to-metal charge transfer electronic band near 500 mm in Tris-HCl buffer. The complexes show binding to Calf Thymus (CT) DNA. Complex 2 showed better binding propensity to the synthetic oligomer poly(dA)center dot poly(dT) than to CT-DNA or poly(dG)center dot poly(dC). All the complexes displayed chemical nuclease activity in the presence of 3-mercaptopropionic acid as a reducing agent and cleaved supercoiled pUC19 DNA to its nicked circular form. They exhibited photo-induced DNA cleavage activity in UV-A light and visible light via a mechanistic pathway that involves the formation of reactive hydroxyl radical species. (C) 2010 Elsevier Inc. All rights reserved.

A FEM-Based Forward Solver for Studying the Forward Problem of Electrical Impedance Tomography (EIT) with A Practical Biological Phantom

A Finite Element Method based forward solver is developed for solving the forward problem of a 2D-Electrical Impedance Tomography. The Method of Weighted Residual technique with a Galerkin approach is used for the FEM formulation of EIT forward problem. The algorithm is written in MatLAB7.0 and the forward problem is studied with a practical biological phantom developed. EIT governing equation is numerically solved to calculate the surface potentials at the phantom boundary for a uniform conductivity. An EIT-phantom is developed with an array of 16 electrodes placed on the inner surface of the phantom tank filled with KCl solution. A sinusoidal current is injected through the current electrodes and the differential potentials across the voltage electrodes are measured. Measured data is compared with the differential potential calculated for known current and solution conductivity. Comparing measured voltage with the calculated data it is attempted to find the sources of errors to improve data quality for better image reconstruction.

A Simple Instrumentation Calibration Technique for Electrical Impedance Tomography (EIT) Using A 16-Electrode Phantom

A simple analog instrumentation for Electrical Impedance Tomography is developed and calibrated using the practical phantoms. A constant current injector consisting of a modified Howland voltage controlled current source fed by a voltage controlled oscillator is developed to inject a constant current to the phantom boundary. An instrumentation amplifier, 50 Hz notch filter and a narrow band pass filter are developed and used for signal conditioning. Practical biological phantoms are developed and the forward problem is studied to calibrate the EIT-instrumentation. An array of sixteen stainless steel electrodes is developed and placed inside the phantom tank filled with KCl solution. 1 mA, 50 kHz sinusoidal current is injected at the phantom boundary using adjacent current injection protocol. The differential potentials developed at the voltage electrodes are measured for sixteen current injections. Differential voltage signal is passed through an instrumentation amplifier and a filtering block and measured by a digital multimeter. A forward solver is developed using Finite Element Method in MATLAB7.0 for solving the EIT governing equation. Differential potentials are numerically calculated using the forward solver with a simulated current and bathing solution conductivity. Measured potential data is compared with the differential potentials calculated for calibrating the instrumentation to acquire the voltage data suitable for better image reconstruction.

Studies on optical absorption and photoluminescenceproperties of Pr3+ and Nd3+ doped mixed alkali chloroborate glasses

This article presents the optical absorption and emission properties of Pr3+ and Nd3+ doped two different mixed alkali chloroborate glass matrices of the type 70B(2)O(3)center dot xLiCl center dot(30 - x)NaCl and 70B(2)O(3)center dot xLiCl center dot(30 - x)KCl (x = 5, 10, 15.20 and 25). The variation of Judd-Ofelt parameters (Omega(2), Omega(4) and Omega(6)), total radiative transition probabilities (A(T)), radiative lifetimes (tau(R)) and emission cross-sections (sigma(p)) with the variation of alkali contents in the glass matrix have been discussed in detail. The changes in the peak wavelengths of the hypersensitive transition and intensity parameters with x are correlated to the structural changes in the host matrix. The estimated radiative lifetimes of certain excited states of Pr3+ and Nd3+ in these two glass matrices are reported. Peak stimulated emission cross-sections (sigma(p)) are reported for the observed emission transitions of Pr3+ and Nd3+ ions. Branching ratios (beta) of the observed emission transitions obtained from the Judd-Ofelt theory are compared with the values obtained from the emission spectra. (C) 2010 Elsevier B.V. All rights reserved.

Ferrocene-Promoted Photoactivated DNA Cleavage and Anticancer Activity of Terpyridyl Copper(II) Phenanthroline Complexes

Ferrocene-appended copper(II) complexes [Cu( Fc-tpy)(B)](ClO4)(2) (1-3) and [Cu(Ph-tpy)(dppz)](ClO4)(2) (4) as control, where Fc-tpy is 4'-ferroceny1-2,2':6',2 ''-terpyridine, Ph-tpy is 4'-pheny1-2,2':6',2 ''-terpyridine, and B is a phenanthroline base, viz., 1,10-phenanthroline (phen, 1), dipyridoquinoxaline (dpq, 2), and dipyridophenazine (dppz, 3), were prepared and structurally characterized, and their DNA binding, photoactivated DNA cleavage activity, and cytotoxic properties were studied [Fe = (eta(5)-C5H4)Fe-11(eta(5)-C5H5)]. Complexes 1 and 3 as hexafluorophosphate salts were structurally characterized by X-ray crystallography. Molecular structures of [Cu(Fc-tpy)(phen)](PF6)(2) (1a) and [Cu(Fc-tpy)(dppz)](PF6)(2)center dot MeCN (3a center dot MeCN) show a distorted square-pyramidal geometry at copper(II), with the Fc-tpy ligand and the phenanthroline base showing respective tridentate and bidentate binding modes. The phenanthroline base exhibits axial-equatorial bonding, while the Fc-tpy ligand binds at the basal plane. The complexes showed quasi-reversible cyclic voltammetric responses near 0.45 and -0.3 V vs SCE in aqueous DMF-0.1 M KCl assignable to the Fc(+)-Fc and Cu(II) Cu(1) redox couples, respectively. The complexes bind to DNA, giving K-b values of 1.4 x 10(4) to 5.6 x 10(5) M-1 in the order 4 similar to 3 > 2 > 1. Thermal denaturation and viscometric titration data suggest groove and/or partial intercalative mode of DNA binding of the complexes. The complexes showed chemical nuclease activity in the presence of 3-mercaptopropionic acid (0.5 mM) or H2O2 (0.25 mM). Complexes 2-4 showed plasmid DNA cleavage activity in visible light, forming (OH)-O-center dot radicals. The Fc-tpy complex 3 showed better DNA photocleavage activity than its Ph-tpy analogue. The ferrocene moiety in the dppz complex 3 makes it more photocytotoxic than the Ph-tpy analogue 4 in HeLa cells.

An Application of 3-D Kinematical Conservation Laws: Propagation of a 3-D Wavefront

Three-dimensional (3-D) kinematical conservation laws (KCL) are equations of evolution of a propagating surface Omega(t) in three space dimensions. We start with a brief review of the 3-D KCL system and mention some of its properties relevant to this paper. The 3-D KCL, a system of six conservation laws, is an underdetermined system to which we add an energy transport equation for a small amplitude 3-D nonlinear wavefront propagating in a polytropic gas in a uniform state and at rest. We call the enlarged system of 3-D KCL with the energy transport equation equations of weakly nonlinear ray theory (WNLRT). We highlight some interesting properties of the eigenstructure of the equations of WNLRT, but the main aim of this paper is to test the numerical efficacy of this system of seven conservation laws. We take several initial shapes for a nonlinear wavefront with a suitable amplitude distribution on it and let it evolve according to the 3-D WNLRT. The 3-D WNLRT is a weakly hyperbolic 7 x 7 system that is highly nonlinear. Here we use the staggered Lax-Friedrichs and Nessyahu-Tadmor central schemes and have obtained some very interesting shapes of the wavefronts. We find the 3-D KCL to be suitable for solving many complex problems for which there presently seems to be no other method capable of giving such physically realistic features.

Hammasröntgentutkimusten termoluminesenssidosimetria

Tavanomaisten hammasröntgenlaitteiden säteilyannoksia valvotaan postitettavien testipakettien ja paikan päällä tehtävien tarkastusten avulla. Säteilyannoksen valvontaan käytetään termoluminesenssidosimetrejä (Thermoluminescence Dosimetry, TLD). Dosimetreissä on TL-materiaalista valmistettuja loistekiteitä, joihin absorboitunut säteilyenergia vapautuu valona materiaalia lämmitettäessä. Prosessissa vapautuvan valon intensiteetti on suoraan verrannollinen absorboituneeseen säteilyannokseen. TLD:llä mitataan rekisteröityjen intraoraalilaitteiden tuottamaa säteilyannosta potilaan posken kohdalla. Säteilyturvakeskus (STUK) ylläpitää rekisteriä ilmoitusvelvollisuuden alaisista hammasröntgenlaitteista. Nyt hammaslaiterekisteriä ollaan uudistamassa siten, että TLD-mittaustulosten käsittely ja annoslaskenta siirtyvät rekisteristä WinTLD-laskentaohjelmaan, jossa on kaikki tarvittavat parametrit annoksen laskemiseksi. Tässä työssä TLD-mittausjärjestelmän kalibrointituloksia analysoitiin vuosilta 1996-2011 ja määritettiin uudelleen laskennassa käytetty energiakorjauskerroin, joka on osa tulevaa WinTLD-konfigurointia. Mittauksissa tarvittavat standardisäteilylaadut (ISO H-laadut) pystytettiin osana työtä. Henkilödosimetrien suorituskykytestauksessa käytetään ISO N-säteilylaatuja. Mirion Technologies (RADOS) käyttää TLD-systeemiä henkilödosimetriassa, ja hammas-TLD on tämän järjestelmän sovellus potilasdosimetriaan. ISO H-laadut otettiin käyttöön, jotta dosimetrien vastetta voitiin ISO N-laatujen tapaan tutkia jatkuvana fotonienergian funktiona Cs-137 ja Co-60 gammasäteilylaatuihin asti ja koska niillä voitiin jäljitellä todellista kliinistä suodatusta. Energiakorjauskerroin kalibroinnissa käytettävän Co-gammasäteilyn ja intraoraalikuvauksissa käytettävän röntgensäteilyn välillä määritettiin uudelleen. Sen arvoksi (yksikkö mGy/mGy) saatiin ISO N-60-laadulla 0,671 ja ISO H-60-laadulla 0,677, jotka ovat numeerisesti hyvin lähellä aikaisemmin määritettyä kerrointa 0,679. Energiakorjauskertoimen epävarmuudeksi saatiin 3,5 % (2std) ja annosmittauksen epävarmuudeksi 7,8 %. Energiavasteiden perusteella dosimetreissä käytetty materiaali on kahdesta vaihtoehdosta MTS-N (LiF:Mg,Ti) eikä MCP-N (LiF:Mg,Cu,P). TLD-järjestelmää voidaan kehittää ja konfiguroida uusien tulosten perusteella, jolloin otetaan käyttöön muun muassa uudelleenmääritetty energiakorjauskerroin. ISO H-säteilylaadut otettiin 22.3.2011 virallisesti käyttöön STUKissa ja niitä käytetään dosimetritestauksessa tarvittaessa suuria annosnopeuksia ja annoksia.

A rationalisation of fatigue thresholds in pearlitic steels using a theoretical model

From a detailed re-examination of results in the literature, the effects of microstructure sizes, namely interlamellar spacing, pearlitic colony size and the prior austentitic grain size on the thresholds for fatigue crack growth (ΔKth) and crack closure (Kcl, th) have been illustrated. It is shown that while interlamellar spacing explicitly controls yield strength, a similar effect on ΔKth cannot be expected. On the other hand, the pearlitic colony size is shown to strongly influence ΔKth and Kcl, th through the deflection and retardation of cracks at colony boundaries. Consequently, an increase in ΔKth and Kcl, th with colony size has been found. The development of a theoretical model to illustrate the effects of colony size, shear flow stress in the slip band and macroscopic yield strength on Kcl, th and ΔKth is presented. the model assumes colony boundaries as potential sites for slip band pile-up formation and subsequent crack deflection finally leading to zig-zag crack growth. Using the concepts of roughness induced crack closure, the magnitude of Kcl, th is quantified as a function of colony size. In deriving the model, the flow stress in the slip band has been considered to represent the work hardened state in pearlite. Comparison of the theoretically predicted trend with the experimental data demonstrates very good agreement. Further, the intrinsic or closure free component of the fatigue threshold, ΔKeff, th is found to be insensitive to colony size and interlamellar spacing. Using a criterion for intrinsic fatigue threshold which considers the attainment of a critical fracture stress over a characteristic distance corresponding to interlamellar spacing, ΔKth values at high R values can be estimated with reasonable accuracy. The magnitude of ΔKth as a function of colony size is then obtained by summing up the average value of experimentally obtained ΔKeff, th values and the predicted Kcl, th values as a function of colony size. Again, very good agreement of the theoretically predicted ΔKth values with those experimentally obtained has been demonstrated.

Plant aspartate transcarbamylase: An affinity chromatographic method for the purification of the enzyme from germinated seedlings

A simple and rapid affinity chromatographic method for the isolation of aspartate transcarbamylase from germinated seedlings of mung bean (Phaseolus aureus) was developed. A partially purified preparation of the enzyme was chromatographed on an affinity column containing aspartate linked to CNBr-activated Sepharose 4B. Aspartate transcarbamylase was specifically eluted from the column with 10 mImage aspartate or 0.5 Image KCl. The enzyme migrated as a single sharp band during disc electrophoresis at pH 8.6 on polyacrylamide gels. Electrophoresis of the sodium dodecyl sulfate-treated enzyme showed two distinct protein bands, suggesting that the mung bean aspartate transcarbamylase was made up of nonidentical subunits. Like the enzyme purified by conventional procedures, this enzyme preparation also exhibited positive homotropic interactions with carbamyl phosphate and negative heterotropic interactions with UMP. This method was extended to the purification of aspartate transcarbamylase from Lathyrus sativus, Eleucine coracona, and Trigonella foenum graecum.

Studies on the 14α-hydroxylation of progesterone in mucor piriformis

Cell-free extracts with high 14?-hydroxylase activity were prepared from induced vegetative cell cultures of Mucor piriformis by grinding in potassium phosphate buffer (0.05 M, pH 8.0) containing glucose (0.25 M), KCl (1 mM), glutathione (1.0 mM) and glycerol (10%). Although the ideal pH for preparing the cell-free extract from vegetative cells was 8.0, the pH optimum of the hydroxylase was found to be 7.6. Microsomes (2.0 mg) prepared from the crude cell-free extract hydroxylated progesterone to 14?-hydroxyprogesterone in not, vert, similar60% yields in 30 min in the presence of NADPH and O2. Microsomes prepared from the uninduced cells did not contain any 14?-hydroxylase activity. The hydroxylase activity was inhibited to a significant extent by CO and p-chloromercuribenzoate whereas moderate inhibition was noticed in the presence of SKF-525A, metyrapone and N-methylmaleimideindicating the possible involvement of the cytochromeP-450 system in the reaction. The membrane bound hydroxylase was solubilized using Triton X-100 and the solubilized fraction contained nearly 35% of the original hydroxylase activity.

Ab initio molecular orbital calculations on ion pair-water complexes of metal halides and oxides

Ab initio MO calculations are performed on a series of ion-molecular and ion pair-molecular complexes of H2O + MX (MX = LiF, LiCl, NaCl, BeO and MgO) systems. BSSE-corrected stabilization energies, optimized geometrical parameters, internal force constants and harmonic vibrational frequencies have been evaluated for all the structures of interest. The trends observed in the geometrical parameters and other properties calculated for the mono-hydrated contact ion pair complexes parallel those computed for the complexes of the individual ions. The bifurcated structures are found to be saddle points with an imaginary frequency corresponding to the rocking mode of water molecules. The solvent-shared ion pair complexes have high interaction energies. Trends in the internal force constant and harmonic frequency values are discussed in terms of ion-molecular and ion-pair molecular interactions.