Rapid screening of solvents and carrier compounds for lactic acid recovery by emulsion liquid extraction and toxicity on Lactobacillus casei (ATCC 11443)

This paper describes a rapid method to identify the best solvent and carrier compound combinations with the highest extraction capability and the lowest microbial toxicity characteristics for product recovery from microbial fermentation. The extraction system has an aqueous phase, and an emulsion phase, which was a blend of sodium carbonate and organic phase [91% (v/v) organic solvent, 5% (v/v or wt/v) carrier compound, and 4% (v/v) surfactant Span 80]. Alamine 336, or tri-n-octylamine in n-heptane; Alamine 336, Alamine 304, or tributyl phosphate in hexane; and Alamine 304 or tributyl phosphate in iso-octane; Alamine 304 or Amberlite in xylene demonstrated high lactic acid extraction. For determination of bacterial toxicity of selected solvent and carrier compounds, Lactobacillus casei subsp. rhamnosus (ATCC 11443) was grown in LAF medium containing one of the selected organic solvent, carrier compound, and Span 80 in 250 ml flask at 37 °C and 125 rpm. Samples were collected regularly during 48 hour incubation, and measured for changes in cell density by absorbance at 620 nm, cell count using a fluorescent dye with flow cytometry, and lactic acid, and glucose concentrations by HPLC. Hexadecane:tributyl phosphate, n-dodecane:tri-n-octylamine, and kerosene:tri-n-octylphosphine oxide demonstrated the least microbial toxicity among the tested blends with excess solvent media. Whereas, hexanes:Alamine 304 and xylenes:Alamine 304 were nontoxic in solvent saturated media.This revised version was published online in October 2005 with corrections to the Cover Date.     

Response Characteristics of Soil Fractal Features to Different Land Uses in Typical Purple Soil Watershed

As a fundamental characteristic of soil physical properties, the soil Particle Size Distribution (PSD) is important in the research on soil moisture migration, solution transformation, and soil erosion. In this research, the PSD characteristics with distinct methods in different land uses are analyzed. The results show that the upper bound of the volume domain of the clay domain ranges from 5.743μm to 5.749μm for all land-use types. For the silt domain of purple soil, the value ranges among 286.852~286.966 μm. For all purple soil land-use types, the order of the volume domain fractal dimensions is D_clay<D_silt<D_sand. However, the values of D_silt and D_sand in the Pinus massoniana Lamb, Robinia pseudoacacia L and Ipomoea batatas are all higher than the corresponding values in the Citrus reticulate Blanco and Setaria viridis. Moreover, in all the land-use types, all of the parameters in volume domain fractal dimension (D_vi) are higher than the corresponding parameter values from the United States Department of Agriculture (D_vi(U)). The correlation study between the volume domain fractal dimension and the soil properties shows that the intensity of correlation to the soil texture and soil organic matter has the order as: D_silt>D_silt(U)>D_sand (U)>D_sand and D_silt>D_silt(U)>D_sand>D_sand(U), respectively. As it is compared with all D_vi, the D_silt has the most significant correlativity to the soil texture and organic matter in different land uses of the typical purple soil watersheds. Therefore, D_silt will be a potential indictor for evaluating the proportion of fine particles in the PSD, as well as a key measurement in soil quality and productivity studies.     

Uncoupling of Bacterioplankton and Phytoplankton Production in Fresh Waters Is Affected by Inorganic Nutrient Limitation

Pelagic bacterial production is often positively correlated, or coupled, with primary production through utilization of autotrophically produced dissolved organic carbon. Recent studies indicate that inorganic N or P can directly limit both bacterial and phytoplanktonic growth. Our mesocosm experiments, with whole communities from mesotrophic Calder Lake, test whether this apparent bacterial-algal coupling may be the result of independent responses to limiting inorganic nutrients. In systems without N additions, numbers of bacteria but not phytoplankton increased 2- to 2.5-fold in response to P fertilization (0 to 2.0 μmol of P per liter); this resulted in uncoupled production patterns. In systems supplemented with 10 μmol of NH₄NO₃ per liter, P addition resulted in up to threefold increases in bacteria and two- to fivefold increases in total phytoplankton biomass (close coupling). P limitation of pelagic bacteria occurred independently of phytoplankton dynamics, and regressions between bacterial abundance and phytoplankton chlorophyll a were nonsignificant in all systems without added N. We describe a useful and simple coupling index which predicts that shifts in phytoplankton and bacterioplankton growth will be unrelated (Δ bacteria/Δ phytoplankton → either + ∞ or - ∞) in systems with inorganic N/P (molar) ratios of <~40. In systems with higher N/P ratios (>40), the coupling index will approach 1.0 and close coupling between bacteria and phytoplankton is predicted to occur.     

The effect of post-lunch napping on mood,reaction time,and antioxidant defense during repeated sprint exercice

To compare the effects of two nap opportunities (20 and 90 min) to countermeasure the transient naturally occurring increased sleepiness and decreased performances during the post-lunch dip (PLD). Fourteen highly trained judokas completed in a counterbalanced and randomized order three test sessions (control (No-nap), 20- (N20) and 90-min (N90) nap opportunities). Test sessions consisted of the running-based anaerobic sprint test (RAST), simple and multiple-choice reaction times (MCRT) and the Epworth sleepiness scale (ESS). From the RAST, the maximum (P_max), mean (P_mean) and minimum (P_min) powers were calculated. Blood samples were taken before and after the RAST to measure the effect of pre-exercise napping on energetic and muscle damage biomarkers and antioxidant defense. N20 increased P_max and P_mean compared to No-nap (p < 0.001, d = 0.59; d = 0.66) and N90 (p < 0.001, d = 0.98; d = 0.72), respectively. Besides, plasma lactate and creatinine increased only when the exercise was performed after N20. Both N20 (p < 0.001, d = 1.18) and N90 (p < 0.01, d = 0.78) enhanced post-exercise superoxide dismutase activity compared to No-nap. However, only N20 enhanced post-exercise glutathione peroxidase activity (p < 0.001, d = 1.01) compared to pre-nap. Further, MCRT performance was higher after N20 compared to No-nap and N90 (p < 0.001, d = 1.15; d = 0.81, respectively). Subjective sleepiness was lower after N20 compared to No-nap (p < 0.05, d = 0.92) and N90 (p < 0.01, d = 0.89). The opportunity to nap for 20 min in the PLD enhanced RAST, MCRT performances, and antioxidant defense, and decreased sleepiness. However, the opportunity of 90 min nap was associated with decreased repeated sprint performances and increased sleepiness, probably because of the sleep inertia.     

Hydration-State Change of Horse Heart Cytochrome c Corresponding to Trifluoroacetic-Acid-Induced Unfolding

We investigate the hydration state of horse-heart cytochrome c (hh cyt c) in the unfolding process induced by trifluoroacetic acid (TFA). The conformation of hh cyt c changes from the native (N) state (2.9 < pH < 6.0) to the acid-unfolded (U_A) state (1.7 < pH < 2.0) to the acid-induced molten globule (A) state (pH ∼1.2). Hydration properties of hh cyt c during this process are measured at 20°C by high-resolution dielectric relaxation (DR) spectroscopy, UV-vis absorbance, and circular dichroism spectroscopy. Constrained water of hh cyt c is observed at every pH as an ∼5-GHz Debye component (DC) (DR time, τ_D ∼30 ps) and its DR amplitude (DRA) is increased by 77% upon N-to-U_A transition, when pH changes from 6.0 to 2.0. Even in the N state, the DRA of the constrained-water component is found to be increased by 22% with decreasing pH from 6.0 to 2.9, suggesting an increase in the accessible surface area of native hh cyt c. Moreover, hypermobile water around native hh cyt c is detected at pH 6.0 as a 19-GHz DC (τ_D ∼ 8.4 ps < τ_DW = 9.4 ps), but is not found at other pH values. The DRA signal of constrained water is found to return to the pH 2.9 (N-state) level upon U_A-to-A transition. Fast-response water (slightly slower than bulk) around A-state hh cyt c is detected at pH 1.2, and this suggests some accumulation of TFA⁻ ions around the peptide chain. Thus, this high-resolution DR spectroscopy study reveals that hh cyt c exhibits significant hydration-state change in the TFA-unfolding process.     

Differences in SOM Decomposition and Temperature Sensitivity among Soil Aggregate Size Classes in a Temperate Grasslands

The principle of enzyme kinetics suggests that the temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition is inversely related to organic carbon (C) quality, i.e., the C quality-temperature (CQT) hypothesis. We tested this hypothesis by performing laboratory incubation experiments with bulk soil, macroaggregates (MA, 250–2000 μm), microaggregates (MI, 53–250 μm), and mineral fractions (MF, <53 μm) collected from an Inner Mongolian temperate grassland. The results showed that temperature and aggregate size significantly affected on SOM decomposition, with notable interactive effects (P<0.0001). For 2 weeks, the decomposition rates of bulk soil and soil aggregates increased with increasing incubation temperature in the following order: MA>MF>bulk soil >MI(P <0.05). The Q₁₀ values were highest for MA, followed (in decreasing order) by bulk soil, MF, and MI. Similarly, the activation energies (E_a) for MA, bulk soil, MF, and MI were 48.47, 33.26, 27.01, and 23.18 KJ mol⁻¹, respectively. The observed significant negative correlations between Q₁₀ and C quality index in bulk soil and soil aggregates (P<0.05) suggested that the CQT hypothesis is applicable to soil aggregates. Cumulative C emission differed significantly among aggregate size classes (P <0.0001), with the largest values occurring in MA (1101 μg g⁻¹), followed by MF (976 μg g⁻¹) and MI (879 μg g⁻¹). These findings suggest that feedback from SOM decomposition in response to changing temperature is closely associated withsoil aggregation and highlights the complex responses of ecosystem C budgets to future warming scenarios.     

High Temperature and Salinity Enhance Soil Nitrogen Mineralization in a Tidal Freshwater Marsh

Soil nitrogen (N) mineralization in wetlands is sensitive to various environmental factors. To compare the effects of salinity and temperature on N mineralization, wetland soils from a tidal freshwater marsh locating in the Yellow River Delta was incubated over a 48-d anaerobic incubation period under four salinity concentrations (0, 10, 20 and 35‰) and four temperature levels (10, 20, 30 and 40°C). The results suggested that accumulated ammonium nitrogen (NH₄ ⁺-N) increased with increasing incubation time under all salinity concentrations. Higher temperatures and salinities significantly enhanced soil N mineralization except for a short-term (≈10 days) inhibiting effect found under 35‰ salinity. The incubation time, temperature, salinity and their interactions exhibited significant effects on N mineralization (P<0.001) except the interactive effect of salinity and temperature (P>0.05), while temperature exhibited the greatest effect (P<0.001). Meanwhile, N mineralization processes were simulated using both an effective accumulated temperature model and a one-pool model. Both models fit well with the simulation of soil N mineralization process in the coastal freshwater wetlands under a range of 30 to 40°C (R² = 0.88–0.99, P<0.01). Our results indicated that an enhanced NH₄ ⁺-N release with increasing temperature and salinity deriving from the projected global warming could have profound effects on nutrient cycling in coastal wetland ecosystems.     

Influence of Electrolytes on the Thicknesses of the Phospholipid Bilayers of Lamellar Lecithin Mesophases

Over a wide range of water contents, aqueous lecithin-water mixtures are mesophases in which lecithin bilayers alternate with water layers. This paper reports on low-angle X-ray diffraction measurements of the effects of electrolytes, at 1.0 N concentration, on the thicknesses of the bilayers in mesophases formed by the synthetic lecithin: 1-octadec-9-enyl-2-hexadecylglycerophosphocholine. With solutions of LiCl, NaCl, Na₂SO₄, KCl, and CsCl, the bilayer thicknesses are less than with pure water. The maximum reduction in bilayer thickness with these electrolytes is about 10% and occurs with mesophases of high content of KCl and CsCl solutions. With HCl solutions the bilayer thicknesses are about 5% greater than with pure water, and with CaCl₂ solutions the bilayer thicknesses are about the same as with pure water. The maximum amount of solution which can be mixed with lecithin before a second, purely aqueous phase is formed is also affected by electrolytes, the order for the various 1.0 N solutions being CsCl = KCl > NaCl > Na₂SO₄ > (pure water) = LiCl > CaCl₂.     

Inactivation of Enteric Adenovirus and Feline Calicivirus by Chlorine Dioxide

Chlorine dioxide (ClO₂) inactivation experiments were conducted with adenovirus type 40 (AD40) and feline calicivirus (FCV). Experiments were carried out in buffered, disinfectant demand-free water under high- and low-pH and -temperature conditions. Ct values (the concentration of ClO₂ multiplied by contact time with the virus) were calculated directly from bench-scale experiments and from application of the efficiency factor Hom (EFH) model. AD40 Ct ranges for 4-log inactivation (Ct_99.99%) at 5°C were >0.77 to <1.53 mg/liter × min and >0.80 to <1.59 mg/liter × min for pH 6 and 8, respectively. For 15°C AD40 experiments, >0.49 to <0.74 mg/liter × min and <0.12 mg/liter × min Ct_99.99% ranges were observed for pH 6 and 8, respectively. FCV Ct_99.99% ranges for 5°C experiments were >20.20 to <30.30 mg/liter × min and >0.68 mg/liter × min for pH 6 and 8, respectively. For 15°C FCV experiments, Ct_99.99% ranges were >4.20 to <6.72 and <0.18 mg/liter × min for pH 6 and 8, respectively. Viral inactivation was higher at pH 8 than at pH 6 and at 15°C than at 5°C. Comparison of Ct values and inactivation curves demonstrated that the EFH model described bench-scale experiment data very well. Observed bench-scale Ct_99.99% ranges and EFH model Ct_99.99% values demonstrated that FCV is more resistant to ClO₂ than AD40 for the conditions studied. U.S. Environmental Protection Agency guidance manual Ct_99.99% values are higher than Ct_99.99% values calculated from bench-scale experiments and from EFH model application.     

Design of antisense oligonucleotides stabilized by locked nucleic acids

The design of antisense oligonucleotides containing locked nucleic acids (LNA) was optimized and compared to intensively studied DNA oligonucleotides, phosphorothioates and 2′-O-methyl gapmers. In contradiction to the literature, a stretch of seven or eight DNA monomers in the center of a chimeric DNA/LNA oligonucleotide is necessary for full activation of RNase H to cleave the target RNA. For 2′-O-methyl gapmers a stretch of six DNA monomers is sufficient to recruit RNase H. Compared to the 18mer DNA the oligonucleotides containing LNA have an increased melting temperature of 1.5–4°C per LNA depending on the positions of the modified residues. 2′-O-methyl nucleotides increase the T_m by only <1°C per modification and the T_m of the phosphorothioate is reduced. The efficiency of an oligonucleotide in supporting RNase H cleavage correlates with its affinity for the target RNA, i.e. LNA > 2′-O-methyl > DNA > phosphorothioate. Three LNAs at each end of the oligonucleotide are sufficient to stabilize the oligonucleotide in human serum 10-fold compared to an unmodified oligodeoxynucleotide (from t_1/2 = ~1.5 h to t_1/2 = ~15 h). These chimeric LNA/DNA oligonucleotides are more stable than isosequential phosphorothioates and 2′-O-methyl gapmers, which have half-lives of 10 and 12 h, respectively.     

Equilibrium studies of lecithin-cholesterol interactions I. Stoichiometry of lecithin-cholesterol complexes in bulk systems

The maximum molar ratio of lecithin:cholesterol in aqueous dispersions has been reported to be 2:1, 1:1, or 1:2. The source of the desparate results has been examined in this study by analyzing (a) the phase relations in anhydrous mixtures (from which most dispersions are prepared) and (b) various methods of preparing aqueous dispersions, with the purpose of avoiding the formation of metastable states that may be responsible for the variability of the lecithin-cholesterol stoichiometry. Temperature-composition phase diagrams for anhydrous mixtures of cholesterol (CHOL) with dimyristoyl (DML) and with dipalmitoyl (DPL) lecithin were obtained by differential scanning calorimetry (DSC). Complexes form with molar ratios for lecithin:CHOL of 2:1 and 1:2; they are stable up to 70°C. When x(CHOL) < 0.33, two phases coexist: complex (2:1) plus pure lecithin; when 0.33 < x(CHOL) < 0.67 complexes (2:1) and (1:2) coexist as separate phases. The corresponding phase diagram in water for these mixtures was determined by DSC and isopycnic centrifugation in D₂O-H₂O gradients. Aqueous dispersions were prepared by various methods (vortexing, dialysis, sonication) yielding identical results except as noted below. The data presented supports the following phase relations. When x(CHOL) < 0.33, two lipid phases coexist: pure lecithin plus complex (2:1) where the properties of the lecithin phase are determined by whether the temperature is below or above T_c, the gel-liquid crystal transition temperature. Therefore, complex (2:1) will coexist with gel state below T_c and with liquid crystal above T_c. The densities follow in the order gel > complex (2:1) > liquid crystal. The density of complex (2:1) is less sensitive to temperature in the range 5°-45°C compared to the temperature dependence for DML and DPL where large changes in density occur at T_c. When x(CHOL) > 0.33, CHOL phase coexists with complex (2:1); anhydrous complex (1:2) is apparently not stable in H₂O. The results are independent of the method and temperature used for preparing the lipid dispersions. However, when dispersions are prepared by sonication or with solvents at T > T_c, an apparent 1:1 complex is formed. Evidence suggests the 1:1 complex is metastable.     

Effect of Pore Size and Porosity on the Biomechanical Properties and Cytocompatibility of Porous NiTi Alloys

Five types of porous Nickel-Titanium (NiTi) alloy samples of different porosities and pore sizes were fabricated. According to compressive and fracture strengths, three groups of porous NiTi alloy samples underwent further cytocompatibility experiments. Porous NiTi alloys exhibited a lower Young’s modulus (2.0 GPa ~ 0.8 GPa). Both compressive strength (108.8 MPa ~ 56.2 MPa) and fracture strength (64.6 MPa ~ 41.6 MPa) decreased gradually with increasing mean pore size (MPS). Cells grew and spread well on all porous NiTi alloy samples. Cells attached more strongly on control group and blank group than on all porous NiTi alloy samples (p < 0.05). Cell adhesion on porous NiTi alloys was correlated negatively to MPS (277.2 μm ~ 566.5 μm; p < 0.05). More cells proliferated on control group and blank group than on all porous NiTi alloy samples (p < 0.05). Cellular ALP activity on all porous NiTi alloy samples was higher than on control group and blank group (p < 0.05). The porous NiTi alloys with optimized pore size could be a potential orthopedic material.     

The natural organosulfur compound dipropyltetrasulfide prevents HOCl-induced systemic sclerosis in the mouse

Introduction

The aim of this study was to test the naturally occurring organosulfur compound dipropyltetrasulfide (DPTTS), found in plants, which has antibiotic and anticancer properties, as a treatment for HOCl-induced systemic sclerosis in the mouse.

Methods

The prooxidative, antiproliferative, and cytotoxic effects of DPTTS were evaluated ex vivo on fibroblasts from normal and HOCl mice. In vivo, the antifibrotic and immunomodulating properties of DPTTS were evaluated in the skin and lungs of HOCl mice.

Results

H₂O₂ production was higher in fibroblasts derived from HOCl mice than in normal fibroblasts (P < 0.05). DPTTS did not increase H₂O₂ production in normal fibroblasts, but DPTTS dose-dependently increased H₂O₂ production in HOCl fibroblasts (P < 0.001 with 40 μM DPTTS). Because H₂O₂ reached a lethal threshold in cells from HOCl mice, the antiproliferative, cytotoxic, and proapoptotic effects of DPTTS were significantly higher in HOCl fibroblasts than for normal fibroblasts. In vivo, DPTTS decreased dermal thickness (P < 0.001), collagen content in skin (P < 0.01) and lungs (P < 0.05), αSMA (P < 0.01) and pSMAD2/3 (P < 0.01) expression in skin, formation of advanced oxidation protein products and anti-DNA topoisomerase-1 antibodies in serum (P < 0.05) versus untreated HOCl mice. Moreover, in HOCl mice, DPTTS reduced splenic B-cell counts (P < 0.01), the proliferative rates of B-splenocytes stimulated by lipopolysaccharide (P < 0.05), and T-splenocytes stimulated by anti-CD3/CD28 mAb (P < 0.001). Ex vivo, it also reduced the production of IL-4 and IL-13 by activated T cells (P < 0.05 in both cases).

Conclusions

The natural organosulfur compound DPTTS prevents skin and lung fibrosis in the mouse through the selective killing of diseased fibroblasts and its immunomodulating properties. DPTTS may be a potential treatment for systemic sclerosis.     

Personality and Dietary Intake – Findings in the Helsinki Birth Cohort Study

Background

Personality traits are associated with health outcomes including non-communicable diseases. This could be partly explained by lifestyle related factors including diet. The personality traits neuroticism, extraversion, openness, agreeableness, and conscientiousness are linked with resilience, meaning adaptability in challenging situations. Resilient people usually comply with favorable health behaviors.

Objective

Our objective was to explore the associations between food and nutrient intake, personality traits and resilience.

Design

A validated semi-quantitative food frequency questionnaire was used to measure diet and the NEO-personality inventory to assess personality in 1681 subjects. Linear regression analysis was used to explore diet-personality associations and cluster analysis to define resilient and non-resilient personality profiles.

Results

Adjusting for age, education and energy intake, and applying Bonferroni corrections, openness in men was associated with higher vegetable (14.9 g/d for 1 SD increase in the personality score, P_Bonf <0.01) and lower confectionery and chocolate (−2.8 g/d, P_Bonf <0.01) intakes. In women, neuroticism was associated with lower fish (−4.9 g/d, P_Bonf <0.001) and vegetable (−18.9 g/d, P_Bonf <0.01) and higher soft drink (19.9 g/d, P_Bonf <0.001) intakes. Extraversion, in women, associated with higher meat (5.9 g/d, P_Bonf <0.05) and vegetable (24.8 g/d, P_Bonf<0.001) intakes, openness with higher vegetable (23.4 g/d, P_Bonf <0.001) and fruit (29.5 g/d, P_Bonf <0.01) intakes. Agreeableness was associated with a lower soft drink (−16.2 g/d, P_Bonf <0.01) and conscientiousness with a higher fruit (32.9 g/d, P_Bonf<0.01) intake in women. Comparing resilient and non-resilient subjects, we found resilience in women to be associated with higher intakes of vegetables (52.0 g/d, P<0.001), fruits (58.3 g/d, P<0.01), fish (8.6 g/d, P<0.01) and dietary fiber (1.6 g/d, P<0.01).

Conclusion

Personality traits are associated with dietary intake and especially subjects with resilient personality profiles had healthier dietary intakes. These associations were stronger in women than in men.     

Self-Subunit Swapping Occurs in Another Gene Type of Cobalt Nitrile Hydratase

Self-subunit swapping is one of the post-translational maturation of the cobalt-containing nitrile hydratase (Co-NHase) family of enzymes. All of these NHases possess a gene organization of <β-subunit> <α-subunit> <activator protein>, which allows the activator protein to easily form a mediatory complex with the α-subunit of the NHase after translation. Here, we discovered that the incorporation of cobalt into another type of Co-NHase, with a gene organization of <α-subunit> <β-subunit> <activator protein>, was also dependent on self-subunit swapping. We successfully isolated a recombinant NHase activator protein (P14K) of Pseudomonas putida NRRL-18668 by adding a Strep-tag N-terminal to the P14K gene. P14K was found to form a complex [α(StrepP14K)₂] with the α-subunit of the NHase. The incorporation of cobalt into the NHase of P. putida was confirmed to be dependent on the α-subunit substitution between the cobalt-containing α(StrepP14K)₂ and the cobalt-free NHase. Cobalt was inserted into cobalt-free α(StrepP14K)₂ but not into cobalt-free NHase, suggesting that P14K functions not only as a self-subunit swapping chaperone but also as a metallochaperone. In addition, NHase from P. putida was also expressed by a mutant gene that was designed with a <β-subunit> <α-subunit> <P14K> order. Our findings expand the general features of self-subunit swapping maturation.     

Contribution of Baicalin on the Plasma Protein Binding Displacement and CYP3A Activity Inhibition to the Pharmacokinetic Changes of Nifedipine in Rats In Vivo and In Vitro

Baicalin purified from the root of Radix scutellariae is widely used in clinical practices. This study aimed to evaluate the effect of baicalin on the pharmacokinetics of nifedipine, a CYP3A probe substrate, in rats in vivo and in vitro. In a randomised, three-period crossover study, significant changes in the pharmacokinetics of nifedipine (2 mg/kg) were observed after treatment with a low (0.225 g/kg) or high (0.45 g/kg) dose of baicalin in rats. In the low- and high-dose groups of baicalin-treated rats, C _max of total nifedipine decreased by 40%±14% (P<0.01) and 65%±14% (P<0.01), AUC_0–∞ decreased by 41%±8% (P<0.01) and 63%±7% (P<0.01), V_d increased by 85%±43% (P<0.01) and 224%±231% (P<0.01), and CL increased by 97%±78% (P<0.01) and 242%±135% (P<0.01), respectively. Plasma protein binding experiments in vivo showed that C _max of unbound nifedipine significantly increased by 25%±19% (P<0.01) and 44%±29% (P<0.01), respectively, and there was a good correlation between the unbound nifedipine (%) and baicalin concentrations (P<0.01). Furthermore, in vitro results revealed that baicalin was a competitive displacer of nifedipine from plasma proteins. In vitro incubation experiments demonstrated that baicalin could also competitively inhibit CYP3A activity in rat liver microsomes in a concentration-dependent manner. In conclusion, the pharmacokinetic changes of nifedipine may be modulated by the inhibitory effects of baicalin on plasma protein binding and CYP3A–mediated metabolism.     

Prooxidant/Antioxidant Balance in Hypoxia: A Cross-Over Study on Normobaric vs. Hypobaric “Live High-Train Low”

“Live High-Train Low” (LHTL) training can alter oxidative status of athletes. This study compared prooxidant/antioxidant balance responses following two LHTL protocols of the same duration and at the same living altitude of 2250 m in either normobaric (NH) or hypobaric (HH) hypoxia. Twenty-four well-trained triathletes underwent the following two 18-day LHTL protocols in a cross-over and randomized manner: Living altitude (P_IO₂ = 111.9 ± 0.6 vs. 111.6 ± 0.6 mmHg in NH and HH, respectively); training “natural” altitude (~1000–1100 m) and training loads were precisely matched between both LHTL protocols. Plasma levels of oxidative stress [advanced oxidation protein products (AOPP) and nitrotyrosine] and antioxidant markers [ferric-reducing antioxidant power (FRAP), superoxide dismutase (SOD) and catalase], NO metabolism end-products (NOx) and uric acid (UA) were determined before (Pre) and after (Post) the LHTL. Cumulative hypoxic exposure was lower during the NH (229 ± 6 hrs.) compared to the HH (310 ± 4 hrs.; P<0.01) protocol. Following the LHTL, the concentration of AOPP decreased (-27%; P<0.01) and nitrotyrosine increased (+67%; P<0.05) in HH only. FRAP was decreased (-27%; P<0.05) after the NH while was SOD and UA were only increased following the HH (SOD: +54%; P<0.01 and UA: +15%; P<0.01). Catalase activity was increased in the NH only (+20%; P<0.05). These data suggest that 18-days of LHTL performed in either NH or HH differentially affect oxidative status of athletes. Higher oxidative stress levels following the HH LHTL might be explained by the higher overall hypoxic dose and different physiological responses between the NH and HH.     

Polarized fluorescence measurements on ordered photosynthetic antenna complexes: Chlorosomes of Chloroflexus aurantiacus and B800-B850 antenna complexes of Rhodobacter sphaeroides

We have used a new and relatively easy approach to study the pigment-organization in chlorosomes from the photosynthetic bacterium Chloroflexus aurantiacus and in B800-850 antenna complexes of the photosynthetic purple bacterium Rhodobacter sphaeroides. These particles were embedded in compressed and uncompressed gels and the polarized fluorescence was determined in a 90° setup. Assuming both a rotational symmetric distribution of the particles in the gel and of the transition dipole moments in the particles, the order parameters <P₂> and <P₄>, describing the orientation of the symmetry axis of the particles with respect to the direction of gel expansion can be determined. Moreover, the direction parameters, describing the orientation of the absorption and emission dipole moments with respect to the symmetry axis of the particles can be obtained.

The value of <P₂> is essential for quantitative interpretation of linear dichroism measurements and usually it is estimated from theoretical approaches, which may lead to incorrect results. For the rod-like chlorosomes the value of <P₂> appears to be the same as predicted by the theoretical approach of Ganago, A. O., M. V. Fok, I. A. Abdourakhmanov, A. A. Solov'ev, and Yu. E. Erokhin (1980. Mol. Biol. [Mosc.]. 14:381-389). The agreement with linear dichroism results, analyzed with this theoretical approach shows that the transition dipole moments are indeed in good approximation distributed in a rotationally symmetric way around the long axis of the chlorosomes. Moreover, it appears those BChl c molecules, which fluoresce, are oriented in the same way with respect to the symmetry axis as the rest of these pigments, with the dipole moments close to parallel to the long axis.

The B800-850 complexes appear to orient like discs, whereas the transition dipoles of the BChl a 800- and 850-nm bands are oriented almost perpendicular to the symmetry axis. These findings are in agreement with the minimal model for these complexes proposed by Kramer, H. J. M., R. van Grondelle, C. N. Hunter, W. H. J. Westerhuis, and J. Amesz (1984. Biochim. Biophys. Acta. 156-165).

The amount of orientation of the particles appears to vary for different gels and it is lower than predicted by the theory of Ganago et al., showing that application of their approach for these particles leads to incorrect interpretations.

The approach that is used in this study allows determination of orientations of those dipole moments, which transfer their excitation energy to the fluorescing species, in contrast to linear dichroism measurements, where the orientations of all absorbing dipole moments are studied. For the polarized fluorescence measurements, the amount of orientation of the particles is determined experimentally, whereas for linear dichroism this amount has to be estimated from theoretical models. The value of <P₂> that can be determined from the fluorescence measurements can, however, also be used for a quantitative interpretation of the linear dichroism results.

     

Effects of Choline on Meat Quality and Intramuscular Fat in Intrauterine Growth Retardation Pigs

The aim of this study was to investigate the effects of choline supplementation on intramuscular fat (IMF) and lipid oxidation in IUGR pigs. Twelve normal body weight (NBW) and twelve intrauterine growth retardation (IUGR) newborn piglets were collected and distributed into 4 treatments (Normal: N, Normal+Choline: N+C, IUGR: I, and IUGR+Choline: I+C) with 6 piglets in each treatment. At 23 d of age, NBW and IUGR pigs were fed basal or choline supplemented diets. The results showed that the IUGR pigs had significantly lower (P<0.05) BW as compared with the NBW pigs at 23 d, 73 d, and 120 d of age, however, there was a slight decreased (P>0.05) in BW of IUGR pigs than the NBW pigs at 200 d. Compared with the NBW pigs, pH of meat longissimus dorsi muscle was significantly lower (P<0.05), and the meat color was improved in IUGR pigs. The malondialdehyde (MDA) levels were significantly decreased (P<0.05), while triglyceride (TG) and IMF contents were significantly higher (P<0.05) in the IUGR pigs than the NBW pigs. IUGR up-regulated the mRNA gene expression of fatty acid synthetase (FAS) and acetyl-CoA carboxylase (ACC). Dietary choline significantly increased (P<0.05) the BW at 120d of age, however, significantly decreased (P<0.05) the TG and IMF contents in both IUGR and NBW pigs. FAS and sterol regulatory element-binding proteins 1 (SREBP1) mRNA gene expressions were increased (P<0.05) while the muscle-carnitine palmityl transferase (M-CPT) and peroxisome proliferators-activated receptorγ (PPARγ) mRNA (P<0.05) gene expressions were decreased in the muscles of the IUGR pigs by choline supplementation. Furthermore, choline supplementation significantly increased (P<0.05) the MDA content as well as the O₂•¯ scavenging activity in meat of IUGR pigs. The results suggested that IUGR pigs showed a permanent stunting effect on the growth performance, increased fat deposition and oxidative stress in muscles. However, dietary supplementation of choline improved the fat deposition via enhancing the lipogenesis and reducing the lipolysis.     

Effect of Sarcoplasmic Reticulum (SR) Calcium Content on SR Calcium Release Elicited by Small Voltage-Clamp Depolarizations in Frog Cut Skeletal Muscle Fibers Equilibrated with 20 mM EGTA

Cut muscle fibers from Rana temporaria (sarcomere length, 3.5–3.9 μm; 14–16°C) were mounted in a double Vaseline-gap chamber and equilibrated with an external solution that contained tetraethyl ammonium– gluconate and an internal solution that contained Cs as the principal cation, 20 mM EGTA, and 0 Ca. Fibers were stimulated with a voltage-clamp pulse protocol that consisted of pulses to −70, −65, −60, −45, and −20 mV, each separated by 400-ms periods at −90 mV. The change in total Ca that entered into the myoplasm (Δ[Ca_T]) and the Ca content of the SR ([Ca_SR]) were estimated with the EGTA/phenol red method (Pape, P.C., D.-S. Jong, and W.K. Chandler. 1995. J. Gen. Physiol. 106:259–336). Fibers were stimulated with the pulse protocol, usually every 5 min, so that the resting value of [Ca_SR] decreased from its initial value of 1,700–2,300 μM to values near or below 100 μM after 18–30 stimulations. Three main findings for the voltage pulses to −70, −65, and −60 mV are: (a) the depletion-corrected rate of Ca release (release permeability) showed little change when [Ca_SR] decreased from its highest level (>1,700 μM) to ∼1,000 μM; (b) as [Ca_SR] decreased below 1,000 μM, the release permeability increased to a maximum level when [Ca_SR] was near 300 μM that was on average about sevenfold larger than the values observed for [Ca_SR] > 1,000 μM; and (c) as [Ca_SR] decreased from ∼300 μM to <100 μM, the release permeability decreased, reaching half its maximum value when [Ca_SR] was ∼110 μM on average. It was concluded that finding b was likely due to a decrease in Ca inactivation, while finding c was likely due to a decrease in Ca-induced Ca release.