31P nmr spin-lattice relaxation studies of deoxyoligonucleotides

Temperature-dependent conformational transitions of deoxyoligonucleotides have been monitored by measuring ³¹P chemical shifts, spin-lattice relaxation times (T₁), and ³¹P-{H} nuclear Overhauser enhancements (NOEs). The measured NOE ranged from 30 to 80%, compared to the theoretical maximum of 124% for a dipolar relaxation mediated by rapid isotropic rotation. The observed 3′-5′ phosphate diester ³¹P T₁ showed a similar temperature dependence over the range 2–75°C for both double- and single-stranded oligonucleotides, and for dinucleotides. The results show that dipole–dipole interactions dominate the internucleotide phosphate relaxation rate in oligonucleotides. The same is true of terminal phosphate groups at low temperature; but at higher temperature another process, possibly due to contamination by paramagnetic ions, becomes dominant. The rotational correlation time τ_R calculated from the dipole–dipole relaxation rate of the internucleotide phosphate in d(pA)₂ at 16°C is τ_R = 5.0 × 10^?10 sec, implying a Stokes radius for isotropic rotation of 7.6 Å. The T₁ and NOE values for the double-helical octanucleotide d(pA)₃pGpC(pT)₃ are consistent with dominance of dipole–dipole relaxation and isotropic rotation of a sphere of radius 14 Å, a reasonable dimension for the double helix. Activation energies for the rotation of dinucleotides range from 4 to 6 kcal/mol, close to the value of 4 kcal/mol expected for isotropic rotation. In order to test the possible effect of internal motion of correlation time τ_G on the results, we considered a model in which the nucleotide chain rotates about the P-O bonds. Comparison of the calculation with our experimental results shows that internal motion with τ_G ? 10^?9 sec, as found from other studies to be present for large nucleic acids, would not influence out T₁ and NOE values enough to be distinguished from isotropic rotation. However, we can conclude that τ_G cannot be as fast as 10^?10 sec, even for dinucleotides.     

Comparative evaluation of quantum theory of nerve excitation

Deliberate evaluation of the quantum theory of nerve excitation is made by comparing it with Hill's theory in fitting the experimental data on threshold-frequency relation, optimum frequency (v₀) for nerve excitation and strength-duration relation. Decrease of v₀ and increase of all the time constants (Hill's λ andk, Wei'sT ₂ and spike durationw) with decreasing temperature are interpreted on the basis of the dipole relaxation timeT ₂ but inexplicable from Hill's theory or any other existing theory. The closeness ofk,T ₂ andw values is explained. A variety of experimental results obtained by others is discussed. Finally, a comparison is made between the Hodgkin-Huxley equations and the quantum theory. Most of the facts (electrical and non-electrical) tend to support the thesis that nerve excitation is a macroscopic expression of quantum transitions of dipoles between energy states.     

The role of lipid and antioxidant exchanges in cell division synchronization (mathematical model)

Cell-cycle synchronization of two diffusecoupled cells has been studied in the framework of the membrane model for the cell division cycle, proposed by Chernavskii et al. (1977). It has been shown semianalytically (using the averaging principle) and by computer stimulation that a) if the duration of theG1-phase (T _G1 ) for two identical cells is comparable with the duration of the remaining cycle (T _S+G2+M ), the lipid (L)-exchange results in a synchronization with phase difference =0. The antioxidant (A)-exchange leads to a phase-locking with =T ₀/2 (whereT ₀ is the cell cycle period; b) ifT _G1 T _S+G2+M (orT _G1 T _S+G2+M ) theL-exchange makes synchronization possible both with =0 and =T ₀/2 while theA-exchange results in phase-locking with confined to the region 0 toT ₀/2; c) for non-identical cells differing in the values of kinetic parameters, the locking band narrows as the population density increases (when some model parameters are close to the bifurcation thresholds). We expect that the cells selected artificially at a definite phase of cycle might maintain the synchronous division for a long time if the lipid exchange between cells were stimulated.     

Vapor–liquid equilibria and thermophysical behavior of the SPC-HW model for heavy water

The vapor–liquid coexistence curve of the simple point charge heavy-water model (SPC-HW), [J. Chem. Phys., 114, 8064–8067 (2001)] is determined by Gibbs Ensemble Monte-Carlo (GEMC) simulation. The estimated critical conditions of the model based on the Wegner-type expansion for the order parameters and the rectilinear diameter are ρ_c = 0.300 g/cc, T _c = 661 K and P _c = 156 bars. The dielectric constant determined by isothermal–isochoric molecular dynamics is underpredicted along the coexistence curve by 29–44% in comparison with the experimental values. The analysis of the orthobaric temperature dependence of the system microstructure, in terms of the three site–site radial distribution functions, indicates that the first coordination numbers for the oxygen–oxygen and the oxygen–deuterium interactions are ～4.3 ± 0.1 and ～1.9 ± 0.1 at T = 300 K, and decrease by 15 and 55%, respectively, at criticality. The dipole–dipole correlation functions show that the orientational order in heavy water is quickly lost beyond the first oxygen–oxygen coordination shell. The model's second virial coefficient is determined by Monte-Carlo integration and used to aid the interpretation of the predicted phase equilibrium results.     

Short-term warming does not affect intrinsic thermotolerance but induces strong sustaining photoprotection in tropical evergreen citrus genotypes

Consequences of warming and postwarming events on photosynthetic thermotolerance (P_T) and photoprotective responses in tropical evergreen species remain elusive. We chose Citrus to answer some of the emerging questions related to tropical evergreen species' P_T behaviour including (i) how wide is the genotypic variation in P_T? (ii) how does P_T respond to short-term warming and (iii) how do photosynthesis and photoprotective functions respond over short-term warming and postwarming events? A study on 21 genotypes revealed significant genotypic differences in P_T, though these were not large. We selected five genotypes with divergent P_T and simulated warming events: T_max 26/20°C (day-time highest maximum/night-time lowest maximum) (Week 1) < T_max 33/30°C (Week 2) < T_max 36/32°C (Week 3) followed by T_max 26/16°C (Week 4, recovery). The P_T of all genotypes remained unaltered despite strong leaf megathermy (leaf temperature > air temperature) during warming events. Though moderate warming showed genotype-specific stimulation in photosynthesis, higher warming unequivocally led to severe loss in net photosynthesis and induced higher nonphotochemical quenching. Even after a week of postwarming, photoprotective mechanisms strongly persisted. Our study points towards a conservative P_T in evergreen citrus genotypes and their need for sustaining higher photoprotection during warming as well as postwarming recovery conditions.     

Frequency dependence of the proton magnetic relaxation in aqueous solutions iof haemoproteins

The longitudinal proton magnetic relaxation times T₁ were measured for ferri (met)-and carbonmonoxy-bovine haemoglobin and equine myoglobin in 0.1 M KH₂PO₄ aqueous solutions near pH 6 at 5°C and 35°C from 1.5- to 60-MHz Larmor frequencies. It is concluded that the correlation time τ_C for the dipole–dipole interaction of electron and nuclear spins is in fact the electron (ferric) spin relaxation time τ_S being close to 1.5 × 10^?10 sec for both metHb and metMb at 5°C. At 35°C the paramagnetic relaxation rates are not determined solely by the relaxation of protons exchanging from the haem pocket with bulk solvent. Hence, τC at 35°C cannot be calculated from the dispersion data obtained at this temperature. The relevance of this for the determination of interspin distances r is discussed.     

Natural Triterpenoids Isolated from Akebia trifoliata Stem Explants Exert a Hypoglycemic Effect via α-Glucosidase Inhibition and Glucose Uptake Stimulation in Insulin-Resistant HepG2 Cells

The inhibition of α-glucosidase activity is a prospective approach to attenuate postprandial hyperglycemia in the treatment of type 2 diabetes mellitus (T2DM). Herein, the inhibition of α-glucosidase by three compounds T₁ – T₃ of Akebia trifoliata stem, namely hederagenin ( T₁ ), 3-epiakebonoic acid ( T₂ ), and arjunolic acid ( T₃ ) were investigated using enzyme kinetics and molecular docking analysis. The three triterpenoids exhibited excellent inhibitory activities against α-glucosidase. T₁ – T₃ showed the strongest inhibition with IC₅₀ values of 42.1±5.4, 19.6±3.2, and 11.2±2.3 μM, respectively, compared to the acarbose positive control (IC₅₀=106.3±8.2). Enzyme inhibition kinetics showed that triterpenoids T₁ – T₃ demonstrated competitive, mixed, and noncompetitive-type inhibition against α-glucosidase, respectively. The inhibition constant (K_i) values were 21.21, 7.70, and 3.18 μM, respectively. Docking analysis determined that the interaction of ligands T₁ – T₃ and α-glucosidase was mainly forced by hydrogen bonds and hydrophobic interactions, which could result in improved binding to the active site of the target enzyme. The insulin resistant (IR)-HepG2 cell model used in this study (HepG2 cells exposed to 10⁻⁷ M insulin for 24 h) and glucose uptake assays showed that compounds T₁ – T₃ had no cytotoxicity with concentrations ranging from 6.25 to 25 μM and displayed significant stimulation of glucose uptake in IR-HepG2 cells. Thus, triterpenoids T₁ – T₃ showed dual therapeutic effects of α-glucosidase inhibition and glucose uptake stimulation and could be used as potential medicinal resources to investigate new antidiabetic agents for the prevention or treatment of diabetes.     

Effects of ovine prolactin on iodide metabolism and thyroid activity in the eel

Summary In the eel, ovine prolactin (oPrl) treatment (0.018 IU/day·g body weight), for 8 to 13 days modifies neither iodide absorption from the water nor excretion, extrathyroidal metabolism and plasma level of iodide.Thyroid activity, evaluated by epithelial cell height, radioiodine uptake and absolute iodide uptake is approximately twice that of controls. However, the amounts of total iodine, thyroxine (T₄) and triiodothyronine (T₃) in thyroid are unaltered by oPrl. Therefore, the decrease of plasma T₄ and the increase of plasma T₃, previously observed in oPrl-treated eels, do not result from a preferential thyroidal secretion of T₃, but only from a stimulation of peripheral conversion of T₄ to T₃. Furthermore, the increased thyroid activity probably originates from a decreased feedback inhibition following the fall of circulating T₄ induced by oPrl.Abbreviations oPrl ovine prolactin - T ₄ Thyroxine - T ₃ 3.5.3 triiodothyronine - TRH thyrotropin releasing hormone - TSH thyroid stimulating hormone - PBI protein bound iodine     

A New Interconnecting Layer of Metal Oxide/Dipole Layer/Metal Oxide for Efficient Tandem Organic Solar Cells

A new metal‐oxide‐based interconnecting layer (ICL) structure of all‐solution processed metal oxide/dipole layer/metal oxide for efficient tandem organic solar cell (OSC) is demonstrated. The dipole layer modifies the work function (WF) of molybdenum oxide (MoO _x ) to eliminate preexisted counter diode between MoO _x and TiO₂. Three different amino functionalized water/alcohol soluble conjugated polymers (WSCPs) are studied to show that the WF tuning of MoO _x is controllable. Importantly, the results show that S‐shape current density versus voltage (J–V) characteristics form when operation temperature decreases. This implies that thermionic emission within the dipole layer plays critical role for helping recombination of electrons and holes. Meanwhile, the insignificant homotandem open‐circuit voltage (V _oc) loss dependence on dipole layer thickness shows that the quantum tunneling effect is weak for efficient electron and hole recombination. Based on this ICL, poly(3‐hexylthiophene) (P3HT)‐based homotandem OSC with 1.20 V V _oc and 3.29% power conversion efficiency (PCE) is achieved. Furthermore, high efficiency poly(4,8‐bis(5‐(2‐ethylhexyl)‐thiophene‐2‐yl)‐benzo[1,2‐b54,5‐b9]dithiophene‐alt alkylcarbonylthieno[3,4‐b]thiophene) (PBDTTT‐C‐T)‐based homotandem OSC with 1.54 V V _oc and 8.11% PCE is achieved, with almost 15.53% enhancement compared to its single cell. This metal oxide/dipole layer/metal oxide ICL provides a new strategy to develop other qualified ICL with different hole transporting layer and electron transporting layer in tandem OSCs.     

Cluster analysis of respiratory time series

We have investigated the respiratory control system with the hypothesis that, although many variables such as minute ventilation (V _I), tidal volume (V _T),breathing period (T _T),inspiratory duration (T _I),and exspiratory duration (T _E),may be observed, the controller functions more simply by manipulating only 2 or 3 of these. Thus, if tidal volume is the only independent variable, T _Ibeing determined by the off-switch threshold, these variables should have very similar time courses. Anesthetized dogs were subjected to CO₂ breathing and carotid sinus perfusion to stimulate both chemoreceptors. The time series of the variables V _I, V _T, T_T, T_Eand T _Ias well as P _{A CO 2}were determined on a breath by breath basis. Derived characteristics of these time series were compared using Cluster Analysis and the latent dimensionality of respiratory control determined by Factor Analysis. The characteristics of the time series clustered into 4 groups: magnitude (of the response), speed, variability and relative change. One respiratory factor accounted for 86% of the variance for the variability characteristics, 2 factors for magnitude (91%) and relative change (85%) and 3 factors for speed (89%). The respiratory variables were analysed for each of the 4 groups of characteristics with the following results: V _Tand T _I clustered together only for the magnitude and relative change characteristics where as T _Tand T _Eclustered closely for all four characteristics. One latent factor was associated with the [T _T-T_E]group and the other usually with P _{A CO 2}.Supported by USPHS 5t01 01919-05, NIH HL 12564 and GM 07033     

Multiple mechanisms generate the resting activity of filiform sensilla in the firebug (<Emphasis Type="Italic">Pyrrhocoris apterus</Emphasis> L.; Heteroptera)

The resting activity was studied in filiform sensilla of the firebug (Pyrrhocoris apterus). Three functional types (T₁, T₂ and T₃) were detected on the abdomen. A resting discharge of nerve impulses is present in all—always in types T₁ and T₂ and occasionally in type T₃. In T₁ the mean rate is 57, in T₂ 3.3 and in T₃ 0.5 imp/s. Shortening the hair length had a negligible effect on the resting discharge, which indicates an intrinsic origin. The resting activity is highly temperature dependent. In T₁, the activation energy was 56.8, in T₂ 84 and in T₃ 61.4 kJ/mol (Q ₁₀: 2.27, 5.6 and 5.5, respectively). Such values are typical for mechano-transduction, suggesting the involvement of the transduction mechanism itself. The destruction of the hair base in T₁ caused halving of the original discharge rate and shifted the discharge to a regular interval mode. The activation energy decreased to 38 kJ/mol. The destruction of the hair bases in T₂ and T₃ completely abolished the discharge. It appears that at least two mechanisms are involved in the generation of the resting activity in T₁ units while only one can be assumed in case of T₂ and T₃.     

Adaptation of homeostatic thermoregulation: comparison of incubating and non-incubating Bantam hens

Summary Incubating and non-incubating Bantam hens were exposed to identical thoracic skin cooling to study the difference between their physiological responses with regard to thermoregulatory adaptation to incubation. Under resting conditions thoracic skin temperature (T _ths) and metabolic heat production (M) were significantly higher in broody than in non-broody hens, indicating a permanently increased conductance of the brood patch. Thoracic skin cooling from 35 to 25 °C decreased T _ths less in broody than in non-broody hens. In broody hens, these coolings induced a large, immediate increase in M, no constriction of brood patch vasculature, and a decrease in colonic temperature (T _c). This decrease in T _c triggered no further increase in M, but induced vasoconstriction in the feet. The coolings induced a smaller increase in M in the non-broody hens, accompanied by pronounced vasoconstriction, and did not affect T _c and foot temperature, T _f. The effects of more severe thoracic skin cooling (between 25 and 15°C) differed much less between non-broody and broody hens. Vasoconstriction of the brood patch also occurred in the latter. It is concluded that in adaptation to incubation the thoracic skin becomes more sensitive, and its input signal becomes stronger for the control of certain effector systems of thermoregulation, allowing a controlled heat transfer to the eggs.Abbreviations BM body mass - M metabolic heat production - T _c colon temperature - T _ths thoracic skin temperature - T _f foot temperature - T _bs back skin temperature - T _stim stimulation temperature - VO₂ oxygen consumption     

Dual Role of Prostaglandins (PGE2) in Regulation of Channel Density and Open Probability of Epithelial Na+ Channels in Frog Skin (R. pipiens)

Prostaglandins are important in signaling pathways involved in modulating the rates of Na⁺ transport in a diverse group of tissues possessing apical membrane epithelial channels. PGE₂ is known to cause either stimulation, inhibition or transient stimulatory changes of Na⁺ transport. We have continued our studies of frog skins that are known to respond to forskolin and PGE₂ with large steady-state increases of transport and have used noninvasive methods of blocker-induced noise analysis of Na⁺ channels to determine their channel densities (N _T ) and open probabilities (P _o ). In the absence of exogenous hormones, baseline rates of Na⁺ transport are especially high in scraped skins (R. pipiens pipiens) studied in the fall of the year. Na⁺ transport was inhibited by indomethacin and by removal of the unstirred layers of the corium (isolated epithelia) alone suggesting that PGE₂ is responsible for the sustained and elevated rates of transport in scraped skins. Changes of transport caused by indomethacin, forskolin or PGE₂ were unquestionably mediated by considerably larger changes of N _T than compensatory changes of P _o . Since cAMP caused no change of P _o in tissues pretreated with indomethacin, PGE₂ appears in this tissue to serve a dual role, increasing the steady state N _T by way of cAMP and decreasing P _o by unknown mechanisms. Despite appreciable PGE₂-related decreases of P _o , the net stimulation of transport occurs by a considerably greater cAMP-mediated increase of N _T . Received: 28 February 1996/Revised: 22 August 1996     

Investigation of the state and dynamics of water in hydrogels of cellulose ethers by 1H NMR spectroscopy

The aim of this work was to study the effect of the type of substituent of the cellulose ethers and the molecular mass on the state and dynamics of water in the respective hydrogels to specify the quantity of adsorbed water on the polymers or, more explicitly, to calculate the average number of water molecules bound to a polymer repeating unit (PRU).¹H NMR relaxation experiments were performed on equilibrated systems of cellulose ether polymers (HEC, HPC, HPMC K4M, and HPMC K100M) with water. In particular, the water proton spinlattice (T ₁) and spin-spin (T ₂) relaxation times were measured in these systems at room temperature. The observed proton NMRT ₁ andT ₂ of water in hydrogels at different cellulose ether concentrations at room temperature were shown to decrease with increasing polymer concentration. The relaxation rate 1/T ₁ is sensitive to the type of polymer substituent but insensitive to the polymer molecular mass. The rate 1/T ₂ appears much less influenced by the polymer substitution. The procedure developed for calculating the amount of water bound per PRU, based on the analysis of theT ₁ andT ₂ data, shows that this amount is the largest for HPC followed by HEC, HP MC K4M, and HPMC K100M. The results correlate well with the degree of hydrophilic substitution of the polymer chains. This NMR analysis deals with a single molecular layer of adsorbed water for the investigated cellulose ether polymers at all concentrations, while the rest of the water in the hydrogel is bulk-like. Therefore, the mesh size of polymer network in the view of a single molecular layer is not effectively changed.     

Triiodo-L-thyronine stimulates glycogen synthesis in rat hepatocyte cultures

The hyperthyroid state is associated with low hepatic glycogen levels, but paradoxically with a high activity of glycogen synthase and low activity of glycogen phosphorylase. We determined the effects of triiodo-L-thyronine (T₃) on glycogen synthesis and glycogen synthase activity in rat hepatocytesin vitro. Culture of rat hepatocytes with T₃ (100 nM–1 M) for 16 h–40 h increases glycogen synthesis from glucose and gluconeogenic precursors. The stimulation of glycogen synthesis by T₃ was associated with an increase in the activity of glycogen synthase and was additive with the long-term effects of insulin but not with the short-term stimulation of glycogen synthesis by insulin. Culture of hepatocytes with T₃ (at concentrations up to 1 M) did not affect the responsiveness of glycogen synthesis to short-term stimulation by insulin but culture with 10 M-T₃ decreased the responsiveness to insulin without affecting the basal rate. It is suggested that the high activity of glycogen synthase in the hyperthyroid state is due to a direct effect of T₃ on the hepatocyte, but the low hepatic glycogen content is probably due to either secondary metabolite and/or endocrine changes or to impaired responsiveness to insulin. T₃ may have an anabolic role in the control of hepatic glycogen storage in the euthyroid postprandial state. (Mol Cell Biol120: 151–158, 1993)Abbreviations T₃ triiodo-L-thyronine     

Effects of temperature and field strength on the NMR relaxation times of Na in frog striated muscle

Pulsed NMR techniques have been applied to the study of the relaxation parameters characterizing ²³Na within frog striated muscle. Experiments were performed at 3°C, 22–24°C and 39°C at a Larmor frequency of 15.7 MHz; at 22–24°C, measurements were obtained both at 15.7 MHz and at 7.85 MHz.As previously reported, only a single spine-lattice relaxation time (T₁) was observed, but both slow (T₂)_I and fast (T₂)_II components of the spin-spin relaxation time were measured. The effect of temperature (θ) upon (1/T₁) was qualitatively similar to that reported for ²³Na in free solution; (θ) did not significantly affect (1/T₂) over the range of temperatures studied. (1/T₂)_I, and to a lesser degreee, (1/T₁) exhibited a modest inverse dependence of doubtful significance on the Larmor frequency.The data are examined within the framework of a simple specific model; a conservative values in assumed for the quadrupolar coupling constant characterizing immobilized intracellular Na⁺. Within this framework, the results suggest that the fraction of bound ions whose molecular tumbling is severely restricted does not exceed some few percent of the total sodium population.     

Thermal stimulation of exposed skin area influences behavioral thermoregulation in pigeons

Summary Pigeons were trained to work instrumentally for thermal reinforcement under ambient heat and cold loads. Facial or breast skin temperature was selectively altered by means of air-perfused thermodes. Facial stimulation with temperatures in the normally occurring range resulted in changes of instrumental response rate, accompanied by stimulus opposing deviations of core temperature (Fig. 5). Breast skin stimulations with the same stimulation amplitudes also affected behavioral thermoregulation, but without changing core temperature. Small temperature changes, as naturally occur at feathered breast skin, had no effect on response rate. Behavioral adjustments occurring within 15 s in response to altering either ambient load temperature or facial temperature locally (Fig. 6), support the hypothesis that the face represents a site driving short term behavioral adjustments to ambient disturbances.Abbreviations RF reinforcement(s) - T _a temporal mean of ambient temperature - T _load load temperature - T _c (colonic) core temperature - T _s skin temperature - T _s(v) ventral skin temperature - T _s(d) dorsal skin temperature - T _ax axilla temperature     

Triiodothyronine Stimulates the Expression of Sucrase–Isomaltase in Caco-2 Cells Cultured in Serum-Free Medium

In a previous study we have shown that triiodothyronine (T₃) added to a serum-free medium supplemented with insulin, transferrin, and selenous acid (ITS) can stimulate Caco-2 cell differentiation. In this study we have focused on the effects of T₃on sucrase activity. The results obtained demonstrate that T₃(50 nM) does not change Caco-2 cell proliferation but enhances sucrase activity from 50 to 80%. Similar increases were observed whether or not insulin was present in the culture medium, showing that there was no synergistic effect between T₃and insulin on sucrase activity. Moreover, T₃acts specifically during the differentiation period since addition of T₃to the defined TS medium before confluency is reached does not stimulate sucrase activity. Sucrase kinetic parameters were evaluated for the first time in Caco-2 cells under various culture conditions. The presence of a single enzyme was verified, with aK_mof about 7 mMand aV_maxaround 20 nmol of substrate hydrolyzed min⁻¹mg⁻¹of protein. Our results showed that T₃did not change the enzyme's affinity for sucrose but doubled theV_max. Moreover, immunoblotting using anti-sucrase–isomaltase (SI) antibodies revealed an approximately twofold increase in the relative amount of SI immunoreactive protein in T₃-stimulated cells compared to untreated cells. Results obtained by both Northern hybridization and RT-PCR amplification showed a significant increase in SI mRNA contents. These results suggest that T₃acts primarily on sucrase expression at the mRNA level.