Topography of nucleic acid helices in solutions. 3. Interactions of spermine and spermidine derivatives with polyadenylic-polyuridylic and polyinosinic-polycytidylic acid helices

The synthesis of spermine derivatives (II), \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_{\rm 2} {\rm R}_{\rm 3} \mathop {\rm N}\limits^ + \left( {{\rm CH}_2 } \right)_3 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} \left( {{\rm CH}_2 } \right)_2 ]_2 \cdot 4{\rm X}^ - $\end{document}, and spermidine derivatives (III), \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_{\rm 2} {\rm R}_{\rm 3} \mathop {\rm N}\limits^ + \left( {{\rm CH}_2 } \right)_4 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} \left( {{\rm CH}_2 } \right)_3 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} {\rm R}_3 \cdot 3{\rm X}^ - $\end{document}, are reported. The effects of these salts on the helix–coil transition of rA–rU and rI–rC helices were examined. Increasing the size of the hydrophobic substituents, R¹, R², and R³ lowers the degree of stabilization of the helical structure. The disproportionation reaction, 2rA–rU→rA–rU₂ + rA occurs readily with salts II and III, especially when the substituents, R₁, R₂, and R₃ are small, i.e., H or Me. Spermine is found to stabilize the rA–rU² and rI–rC helices to approximately the same extent; however, large differences between the degree of stabilization of rA–rU₂ and rI-rC helices are observed when the substituents R₁, R₂, and R₃ are large hydrophobic groups. Similar results are also obtained for the spermidine series. Finally, differences in the interactions of the salts II and III with rA–rU₂ and rI–rC helices suggest that the latter helix is denser.     

Topography of nucleic acid helices in solutions. The interaction specificities of optically active amino acid derivatives

The synthesis and interactions of the d- and l-enantiomers of the amino acid amide derivatives [Formula: see text] (I) and lysyl dipeptides [Formula: see text] (II) with poly rI.poly rC, poly rA.poly rU and calf thymus DNA is reported. The following results were found. (1) The degree of stabilization of the helices as measured by the T(m) (;melting' temperature) of the helix-coil transition was dependent on the nature of the amino acid. (2) For the poly rI.poly rC helix, the l-enantiomers of salts (I) and (II) stabilized more than the d-enantiomers. The same was true for calf thymus DNA in the presence of salts (II) and for poly rA.poly rU in the presence of salts (II) and the proline derivatives of salts (I). (3) As R increased in size and became more apolar, the amount of stabilization of the poly rI.poly rC helix in the presence of salts (I) decreased. On the other hand, the amount of stabilization increased with more polar substituents. An attempt was then made to determine whether the difference in stabilization of the double-stranded helices at the T(m) in the presence of the l- and d-enantiomers of salts (I) is due to the interaction with the helix, the random coil or both. A new method was developed for determining the binding of small ions to polyions that involves a competition between an insoluble polystyrene ion-exchange resin and the soluble polyion for the counterion. Dissociation constants are obtained for the complexes of single- and double-stranded helices with the salts (I). The results are illuminating and indicate that with certain helices, i.e. poly rA.poly rU, the interactions of salts (I) with the single strands may not be ignored. It is concluded that the high optical specificity found in Nature, i.e. d-ribose in nucleic acids and l-amino acids in proteins, cannot be attributed solely to monomer-polymer interactions described by Gabbay (1968).     

Morphometric diffusing capacity and functional anatomy of the book lungs in the spider Tegenaria spp. (Agelenidae)

The presence of both book lungs and a tracheal system in many spiders raises the question of the functional significance of this double respiratory system. The present physiological and morphometric study of the house spider (Tegenaria spp.) reveals that the diffusing capacity (Dto₂) of the lungs alone suffices during rest and following exercise to meet measured rates of oxygen consumption (\documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm.} $\end{document}o₂) at driving pressures (ΔPto ₂) similar to those calculated for vertebrate lungs. During moulting ΔPto ₂ may rise to more than double the vertebrate values, implying the possible insufficiency of book lungs during this critical life phase. Resting \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ is greatest (92 mm³/h · g) during the early morning and lowest (66 mm³/h · g) near midday: during moulting \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ rises to 278.7 mm³/h · g. In spiders recovering from exercise \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ is consistently greater than during rest: neither value is significantly reduced by blockage of the tracheal stigmas. Regression calculations of morphometric values for a hypothetical 100-mg Tegenaria yield a total lung volume of 0.578 mm³, a pulmonary surface area of 69.8 mm², and a surface-to-volume ratio of 120.89 mm²/mm³. In spite of the similar thickness of the chitinous and hypodermal components of the air-hemolymph barrier (each ca. 0.2 μm in nonmoulting animals), the low permeability of chitin for oxygen makes this layer the greater barrier to diffusion. For a 100-mg specimen Dto₂ is 3.5 mm³/h · torr: similar to that of a turtle (Pseudemys) on a gram-body weight basis.     

Transient method for proposing the mechanisms of reactions over immobilized enzymes

The transient response method is introduced to elucidate the mechanism of reaction over immobilized enzyme. Glucose oxidation over the glucose oxidase that was immobilized on ion-exchange resin using glutaraldehyde as a linking agent is selected as an example here. The transient responses of a fixed-bed reactor to step increases and decreases in glucose, oxygen, and gluconolactone feed concentrations have been monitored and interpreted. From some responses, we have found that gluconolactone is formed in the reaction of glucose with adsorbed oxygen, while hydrogen peroxide is formed in the reaction of oxygen with adsorbed glucose. Combining all information from interpreting the responses with the literature, a mechanistic picture can be obtained as follows: \documentclass{article}\pagestyle{empty}\begin{document}$$ \begin{array}{*{20}c} {E_{{\rm ox}} + G \to E_{{\rm red}} GL} \\ {E_{{\rm red}} GL \to E_{{\rm red}} + GL} \\ {E_{{\rm red}} + {\rm O}_2 \to E_{{\rm ox}} {\rm H}_2 {\rm O}_2 } \\ {E_{{\rm ox}} {\rm H}_2 {\rm O}_2 \to E_{{\rm ox}} + {\rm H}_2 {\rm O}_2 } \\ \end{array} $$\end{document}.     

Influence of anions on metal adsorption by Rhizopus arrhizus biomass

The presence of anions in solution was found to inhibit the uptake of La(3+), Cd(2+), Pb(2+), UO(2+) (2), and Ag(+) by Rhizopus arrhizus biomass. The effects ranged from total inhibition of Cd(2+) and Pb(2+) uptake at equimolar concentrations of EDTA to no change in uptake of La(3+) or UO(2+) (2) at 12-fold molar excesses of Cl(-) or CO(2-) (3). No anion was found to enhance metal uptake levels, and the degree of inhibition generally followed the series: \documentclass{article}\pagestyle{empty}\begin{document}$${\rm EDTA } \ge \ge {\rm SO}_{;{;{;{\rm 4} } } };{{\rm 2} - } \ge {\rm Cl}; - \ge {\rm PO}_{;{;{;{\rm 4} } } };{{\rm 3} - } \ge {\rm glutamate} \ge {\rm CO}_{;{;{\rm 3} } };{{\rm 2} - } $$\end{document} The chemical equilibrium model REDEQL2 was adapted to treat metal uptake by R. arrhizus biomass and used to predict the effects of anions in solution. Comparisons with the experimental results are made and discussed in light of the assumptions underlying the model.     

Modellierung der enzymatischen Cellobiosehydrolyse – Vergleich linearer und nichtlinearer Parameterbestimmung

Experimental kinetic data (initial rate and high conversion) on the hydrolysis of cellobiose by 1,4-β-glucosidace (Gliocladium sp.) have been analysed and a competitive inhibition by glucose has been proposed. The determination of kinetic parameters from integral data is based upon algorithms for non-linear optimization and numerical integration. The values of kinetic constants \documentclass{article}\pagestyle{empty}\begin{document}$(v_{\max } = 1.02\frac{{\mu {\rm M}_{{\rm glucose}} }}{{{\rm mg}_{{\rm protein}} \cdot \min }},K_M = 2.6{\rm mM/l, and }K_P = 1.2{\rm mM/l)}$\end{document} agree well with the initialrate results. An important distinction is the confidence limit of parameters. Linear regression analysis shows a virtual accuracy and can lead to wrong conclusions.     

Kinetics of ethanol inhibition in alcohol fermentation

The inhibitory effect of ethanol on yeast growth and fermentation has been studied for the strain Saccharomyces cerevisiae ATCC No. 4126 under anaerobic batch conditions. The results obtained reveal that there is no striking difference between the response of growth and ethanol fermentation. Two kinetic models are also proposed to describe the kinetic pattern of ethanol inhibition on the specific rates of growth and ethanol fermentation: \documentclass{article}\pagestyle{empty}\begin{document}$$\begin{array}{*{20}c} {\frac{{\mu _i }}{{\mu _0 }} = 1{\rm } - {\rm }\left( {\frac{P}{{P_m }}} \right);\alpha } \hfill & {\left( {{\rm for}\ {\rm growth}} \right)} \hfill \\ {\frac{{\nu _i }}{{\nu _0 }} = 1{\rm } - {\rm }\left( {\frac{P}{{P'_m }}} \right);\beta } \hfill & {\left( {{\rm for}\ {\rm ethanol}\ {\rm production}} \right)} \hfill \\ \end{array}$$\end{document} The maximum allowable ethanol concentration above which cells do not grow was predicted to be 112 g/L. The ethanol-producing capability of the cells was completely inhibited at 115 g/L ethanol. The proposed models appear to accurately represent the experimental data obtained in this study and the literature data.     

Sequence and age of eruption of deciduous dentition in the baboon (Papio sp)

A detailed eruption sequence and associated age of eruption for deciduous dentition in baboons (Papio sp) are presented in this paper. The sequence was determined by evaluation and comparison of the number and kinds of teeth present in nine age cohorts comprising the study sample of 88 males and 87 females who ranged in age from birth to 763 days. Eruption was assessed visually as present or absent. Several statistical methods used to derive the ages associated with the eruption sequence are described. The basic eruption sequence in the sample population is: i¹ i₁, i₂, i², \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm c}\limits_{\rm -} {\rm,}\mathop {\rm c}\limits^{\rm -} $\end{document} m¹ (m², m₂), M₁, M¹. Both sexes show the same pattern, with the exception of the second deciduous molar, where males show a sequence of m₂, m₂, while females show the opposite. Posterior dentition shows the greatest gender-specific variation in average age of eruption.     

Optical anisotropy of polypeptide chains

Optical anisotropies γ² of N-t-butylacetamide (tBA), N-Methylacetamide (MA), and N, N-dimethylacetamide (DMA) have been determined from the Rayleigh ratios for depolarzed scattering by dilute solutions of the amides in p-dioxane. Traceless optical polarizability tensors \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document} for the amides are derived from these results in conjunction with the Kerr constant for tBA determined by LeGèvre and co-workers. It is shown that the tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document}_i for the glycyle unit in a polypeptide chain may be identified with \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document}MA . Methods for deriving corresponding tensors for other peptide units are indicated and the traceless polarizability tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document} for a polypeptide chain in any specified configuration is formulated.     

Viscosity of heparin as a function of dielectric constant and of desulfation

The effect of dielectric constant (D) of the solvent on the viscosity of heparin was examined using the relation \documentclass{article}\pagestyle{empty}\begin{document}$ \eta _{{\rm sp}} /c = [\eta ]_\infty (1 + k/\sqrt c) $\end{document}, where [η]_∞ is the shielded intrinsic viscosity obtained by extrapolating \documentclass{article}\pagestyle{empty}\begin{document}$ \eta _{{\rm sp}} /c\,{\rm vs}{\rm . }\,1/\sqrt c ) $\end{document} to infinite concentration, and k is an interaction parameter independent of the dielectric constant of the solvent. This equation was previously reported by the authors⁹ for describing the reduced viscosities of strong polyelectrolytes in salt-free polar solvents. It was found that the [η]_∞ of heparin increases linearly with increasing dielectric constant of the solvent whereas the k values were, within experimental error, independent of D in the range 54.7 < D < 93.2 examined. Graded hydrolysis of heparin from its acid form (heparinic acid) at 57°C resulted in samples of varying degree of desulfation with corresponding decrease in biological activity. It was found that both [η]_∞ and k decrease with increasing desulfation.     

Interaction-induced electric (hyper)polarizability in the dihydrogen–neon pair: basis set and electron correlation effects

We investigated the interaction (hyper)polarizability of neon–dihydrogen pairs by performing high-level ab initio calculations with atom/molecule-specific, purpose-oriented Gaussian basis sets. We obtained interaction-induced electric properties at the SCF, MP2, and CCSD levels of theory. At the CCSD level, for the T-shaped configuration, around the respective potential minimum of 6.437 a₀, the interaction-induced mean first hyperpolarizability varies for 5?<? R/a₀?<?10 as

$$ \left[{\overline{\beta}}_{\mathrm{int}}(R)\hbox{-} {\overline{\beta}}_{\mathrm{int}}\left({R}_{\mathrm{e}}\right)\right]/{e}^3{a_0}^3{E_{\mathrm{h}}}^{-2}=-0.91\left(R\hbox{-} {R}_{\mathrm{e}}\right)+0.50{\left(R\hbox{-} {R}_{\mathrm{e}}\right)}^2\hbox{--} 0.13{\left(R\hbox{-} {R}_{\mathrm{e}}\right)}^3+0.01{\left(R\hbox{-} {R}_{\mathrm{e}}\right)}^4. $$

Again, at the CCSD level, but for the L-shaped configuration around the respective potential minimum of 6.572 a₀, this property varies for 5?<? R/a₀?<?10 as

$$ \left[{\overline{\beta}}_{\mathrm{int}}(R)\hbox{-} {\overline{\beta}}_{\mathrm{int}}\left({R}_{\mathrm{e}}\right)\right]/{e}^3{a_0}^3{E_{\mathrm{h}}}^{-2}=-1.33\left(R\hbox{-} {R}_{\mathrm{e}}\right)+0.75{\left(R\hbox{-} {R}_{\mathrm{e}}\right)}^2-0.20{\left(R\hbox{-} {R}_{\mathrm{e}}\right)}^3+0.02{\left(R\hbox{-} {R}_{\mathrm{e}}\right)}^4. $$

Open image in new window

Graphical Abstract Interaction-induced mean dipole polarizability (\( \overline{a} \)) for the T-shaped configuration of H₂–Ne calculated at the SCF, MP2, and CCSD levels of theory

     

The humus layer determines SO<Subscript>4</Subscript><Superscript>2−</Superscript> isotope values in the mineral soil

Biocycling of sulfur (S) has been proposed to play an important role in the recovery of ecosystems following anthropogenic S deposition. Here, we investigated the importance of the humus layer in the biocycling of S in three forested catchments in the Gårdsjön area of southwestern Sweden with differing S inputs and S isotope signature values. These experimental sites consisted of two reference catchments and the Gårdsjön roof experiment catchment (G1), where anthropogenic deposition was intercepted from 1991 until May 2002 by a roof placed over the entire catchment area. Under the roof, controlled levels of deposition were applied, using a sprinkler system, and the only form of S added was marine SO₄²⁻ with a δ of +19.5‰.We installed ion exchange resin bags at the interface between the humus layer and mineral soil at each of the catchments to collect SO₄²⁻ passing through the humus. The resin bags were installed on four occasions, in 1999 and 2000, covering two summer and two winter periods. The ions collected by each bag during these sampling periods were then eluted and their δ values and SO₄²⁻ concentrations determined. The most striking result is that the average δ value in the resin bags was more than 12‰ lower compared to that of the sprinkler water in the G1 roof catchment. There was no increasing trend in the isotope value in the resin bag SO₄²⁻ despite that the roof treatment has been on-going for almost 10 years; the average value for all resin bags was +7.1‰. The highest δ values found in the G1 roof catchment were between +11‰ and +12‰. However, these values were all obtained from resin bags installed at a single sampling location. Throughfall and resin bag δ values were more similar in the two reference catchments: about +7.5‰ in both cases. There was, however, an increase in resin bag δ values during the first winter period, from about +7‰ to +9‰. The resin bag δ value was linearly and positively related (r² = 0.26, p < 0.001) to the amount of SO₄²⁻ extracted from the resin bags, if relatively high amounts (>50 mmol m⁻²) were excluded. High amounts of resin bag SO₄²⁻ seemed to be related to groundwater inputs, as indicated by the δ value. Our results suggest that rapid immobilization of SO₄²⁻ into a large organic S pool may alter the S isotope value and affect the δ values measured in the mineral soil and runoff.     

Spectroscopic studies of chimeric DNA-RNA and RNA 29-base intramolecular triple helices.

Fourier transform infrared (FTIR), UV absorption and exchangeable proton NMR spectroscopies have been used to study the formation and stability of two intramolecular pH-dependent triple helices composed by a chimeric 29mer DNA-RNA (DNA double strand and RNA third strand) or by the analogous 29mer RNA. In both cases decrease of pH induces formation of a triple helical structure containing either rU*dA.dT and rC+*dG.dC or rU*rA.rU and rC+*rG.rC triplets. FTIR spectroscopy shows that exclusively N-type sugars are present in the triple helix formed by the 29mer RNA while both N- and S-type sugars are detected in the case of the chimeric 29mer DNA-RNA triple helix. Triple helix formation with the third strand RNA and the duplex as DNA appears to be associated with the conversion of the duplex part from a B-form secondary structure to one which contains partly A-form sugars. Thermal denaturation experiments followed by UV spectroscopy show that a major stabilization occurs upon formation of the triple helices. Monophasic melting curves indicate a simultaneous disruption of the Hoogsteen and Watson-Crick hydrogen bonds in the intramolecular triplexes when the temperature is increased. This is in agreement with imino proton NMR spectra recorded as a function of temperature. Comparison with experiments concerning intermolecular triplexes of identical base and sugar composition shows the important role played by the two tetrameric loops in the stabilization of the intramolecular triple helices studied.     

Quantifying Lipari–Szabo modelfree parameters from 13CO NMR relaxation experiments

It is proposed to obtain effective Lipari–Szabo order parameters and local correlation times for relaxation vectors of protein ¹³CO nuclei by carrying out a ¹³CO-R₁ auto relaxation experiment, a transverse CSA/dipolar cross correlation and a transverse ¹³CO CSA/¹³CO–¹⁵N CSA/dipolar cross correlation experiment. Given the global rotational correlation time from ¹⁵N relaxation experiments, a new program COMFORD (CO-Modelfree Fitting Of Relaxation Data) is presented to fit the ¹³CO data to an effective order parameter , an effective local correlation time and the orientation of the CSA tensor with respect to the molecular frame. It is shown that the effective is least sensitive to rotational fluctuations about an imaginary axis and most sensitive to rotational fluctuations about an imaginary axis parallel to the NH bond direction. As such, the information is fully complementary to the ¹⁵N relaxation order parameter, which is least sensitive to fluctuations about the NH axis and most sensitive to fluctuations about the axis. The new paradigm is applied on data of Ca²⁺ saturated Calmodulin, and on available literature data for Ubiquitin. Our data indicate that the order parameters rapport on slower, and sometimes different, motions than the ¹⁵N relaxation order parameters. The CO local correlation times correlate well with the calmodulin’s secondary structure. Electronic Supplementary Material Supplementary material is available to authorized users in the online version of this article at .     

Conformational properties of methionine homo-oligopeptides in solution

A conformational analysis was carried out in solution on a series of L -methionine oligomers having the general formula \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm BOC\rlap{--} (L - Met\rlap{--})}_n {\rm OMe (}n = 2 - 7)$\end{document}. We examined these oligopeptides in TFE, HFIP, EG, and mixed organic–water media. The critical size for helix formation was found to be seven residues in TFE, whereas the β-associated structure appears at the pentamer in EG and TFE–water (20 : 80, v/v). In HFIP, however, the oligomers exist essentially in an unordered conformation.     

Destabilization of the secondary structure of RNA by ribosomal protein S1 from escherichia coli

S1 is an acidic protein associated with the 3′ end of 16S RNA; it is indispensable for ribosomal binding of natural mRNA. We find that S1 unfolds single stranded stacked or helical polynucleotides (poly rA, poly rC, poly rU). It prevents the formation of poly (rA + rU) and poly (rI + rC) duplexes at 10–25 mM NaCl but not at 50–100 mM NaCl. Partial, salt reversible denaturation is also seen with coliphage MS2 RNA, $\text{E. coli}$ rRNA and tRNA. Generally, only duplex structures with a Tm greater than about 55° are formed in the presence of S1. The protein unfolds single stranded DNA but not poly d(A·T).     

Self-thinning lines of organs and aboveground parts based on allometric relationships in overcrowded Pinus densiflora stands

The allometric relationships of mean tree height [`(H)]( μ [`(w)]_x^q ) \bar{H}( \propto \bar{w}_{\rm {x}}^{\theta } ) and of organ mass density [`(d)]<sub>x</sub> ( μ [`(w)]_x^d ) \bar{d}_{\rm {x}} ( \propto \bar{w}_{\rm {x}}^{\delta } ) to mean organ mass [`(w)]_x \bar{w}_{\rm {x}} were studied in self-thinning Pinus densiflora Sieb. et Zucc. stands. Tree height increased significantly with increasing mean mass of organs and aboveground parts on log–log coordinates. The value of the allometric constant θ ranged from 0.2878 to 0.3349. On the other hand, the constant value δ was not significantly different from zero, except for leaves. The value of the allometric constant δ ranged from −0.2926 to 0.0120. According to Weller’s allometric model, the slope of the self-thinning line was calculated from the allometric constants θ and δ. The thinning slope was estimated to be −1.51 in stem, −1.39 in branches, −1.00 in leaf and −1.41 in aboveground parts, respectively. Mass density was high in stem, medium in branches and low in leaves. Mean leaf mass density decreased significantly with decreasing stand density on log–log coordinates, which could be interpreted as indicating the importance of the constant final leaf biomass in overcrowded P. densiflora stands. The self-thinning exponents of branch, stem and aboveground parts were not significantly different from 3/2, which indicated that the 3/2 power law of self-thinning holds for stem mass, branch mass and aboveground mass in overcrowded P. densiflora stands.