Evaluation of the BRAHMS KRYPTOR thyroglobulin "mini-recovery" test in thyroid healthy subjects

The aim of the work was to compare the automated thyroglobulin (Tg) assay on the automated BRAHMS KRYPTOR platform (hTG KRYPTOR) to the established BRAHMS Tg Plus immunoradiometric assay for the measurement of Tg levels and regular Tg recovery rates and to assess a recovery test using a low Tg concentration of 10 μg/l ("mini-recovery") in samples with a native Tg level of <10 μg/l. Tg levels and recovery rates, as well as the mini-recovery, were determined in 208 serum samples from thyroid-healthy patients using both assays. The reference ranges for the Tg-Plus assay are 2.0-51.0 μg/l for Tg levels and 81.5-108% for recovery rates at 100 μg/l. The reference ranges for hTG KRYPTOR are 2.4-47.8 μg/l for Tg, 83.3-110.4% for a conventional recovery with 80 μg/l in Tg levels ≥ 10.0 μg/l (n=121) and 94.4-122.9% for the mini-recovery with Tg <10.0 μg/l (n=87). The correlation between the Tg-Plus and hTG KRYPTOR is excellent for Tg (r2=0.95; p<0.001), but not significant for recovery rates. Tg levels determined using the KRYPTOR Tg assay are clinically comparable to the conventional Tg-Plus assay. New features of the KRYPTOR assay such as the ability to perform a "mini-recovery" still require further study before clinical use.     

Dilution of solar radiation through "culture" lamination in photobioreactor rows facing south-north: a way to improve the efficiency of light utilization by cyanobacteria (Arthrospira platensis)

Efficient utilization of solar radiation for the photoautotrophic production of cyanobacterium biomass was achieved, using small pipes (ID = 0.01 m) arranged in rows in two photobioreactors facing south-north. A high Arthrospira yield of 47.7 g m(-2) (installation area) d(-1) was attained under outdoor conditions in a tubular undulating row photobioreactor (TURP-10r). During the summer, under a semicontinuous culture regime, the optimal biomass concentration (OBC) in TURP-5r was 6.0 g L(-1): it was 5.0 g L(-1) in TURP-10r. These OBCs made it possible to produce a biomass output rate of 2.7 +/- 0.2 g L(-1) d(-1) in the former and 2.1 +/- 0.1 g L(-1) d(-1) in the latter. When Arthrospira was grown at a preset dilution rate (0.3 d(-1)), sunrise cell density (SrCD) variations were not proportional to the drop of solar radiation. The SrCD was comparatively high at high solar radiation and decreased abruptly with decreasing solar radiation. There was a tendency to stabilize at low solar radiation. In both photobioreactors, the chlorophyll content of the Arthrospira biomass (% of the dry weight) was higher at sunrise than at sunset. A comparison of the chlorophyll biomass content in the TURPs showed no significant differences. Night biomass losses were very high (> 30% of the daylight productivity) when the culture temperature was kept constant at 31 +/- 1.0 degrees C: these losses fell to < 20% of the daylight productivity, when the night temperature of the cultures decreased according to the environmental temperature. Dilution of solar radiation was carried out using two quasi-laminated bioreactors. The rows of S-N facing bioreactors showed a very high growth yield in TURP-10r [about 2.1g (d.w.) MJ(-1)]. In TURP-10r, the high photic ratio (R(f) = 6), the high surface-to-volume ratio (S(ill)/V = 400 m(-1)) and the S-N facing of the rows (better than an E-W orientation) allowed for good results.     

Electron spin resonance spectrum of species “X” which may function as the primary electron acceptor in Photosystem I of green plant photosynthesis

Purified Photosystem I particles from spinach when reduced with 10 mM dithionite at pH 9 exhibited a 50% light reversible-ESR Signal 1 (P-700⁺) at about 10 K. It was possible to show by signal-averaging techniques that a light-reversible ESR spectrum concomitant with the reversible Signal 1 can be observed with approximate principal g factors at g = 2.07, g = 1.86 and g = 1.75.     

Survival of <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> in Wilted and Additive-Treated Grass Silage

Grass was field-dried to 3 different dry matter (DM) levels (200, 430 and 540 g/kg) and inoculated with 10⁶–10⁷ cfu/g of a Listeria monocytogenes strain sharing a phagovar occasionally involved in food-borne outbreaks of listeriosis. Formic acid (3 ml/kg) or lactic acid bacteria (8·10⁵/g) with cellulolytic enzymes were applied only to forages with low and intermediate DM levels. Forages were ensiled in laboratory silos (1700 ml) and were stored at 25°C for 30 or 90 days. After 90 days of storage, L. monocytogenes could not be detected in any silo, except one with the high dry matter grass without additive. After 30 days of storage, between 10² and 10⁶ cfu L. monocytogenes/g silage were isolated from the untreated silages. Increasing the DM content from 200 to 540 g/kg did not reduce listeria counts possibly because of the lower production of fermentation acids (higher pH). In silages treated with additives, counts of L. monocytogenes were always lower than in silages without additive. In wet silages (DM 200 g/kg) both additives were effective, but in the wilted silages (DM 430 g/kg) only the bacterial additive reduced listeria counts below detection level. Listeria counts were highly correlated to silage pH (r = 0.92), the concentration of lactic acid (r = -0.80) and the pooled amount of undissociated acids (r = -0.83).     

A novel electron paramagnetic resonance signal of "oxygenated" cytochrome c oxidase.

It had been observed previously that a pair of transient EPR resonances (g = 1.78 and 1.69) appears within less than 5 ms on reoxidation of reduced cytochrome c oxidase by O2. Since the location of other lines that are part of the same signal was not known, the quantity of the paramagnetic species involved, and thus the significance of the observed resonances, remained questionable. We have now found a broad resonance at g = 5 which is obviously associated with those at g = 1.78 and 1.69. The width of the signal (approximately 250 mT) at the observed intensity suggests that it represents a significant fraction of one of the components of the enzyme. The signal disappears within less than 5 ms on addition of cyanide or sulfide but only within several hundred milliseconds after addition of ferrocytochrome c. This behavior suggests that it originates from the a3 component of the enzyme. It is suggested that the species represented in the signal is either identical with or part of what has been named collectively the "oxygenated" form and recently described "activated" forms of the enzyme. On reoxidation of reduced oxidase with oxygen enriched 90% in 17O, no change of signal shape was seen.     

Dual-mode EPR spectrometry of O2-pulsed cytochrome c oxidase

O2-activated bovine heart cytochrome c oxidase has been examined by dual-mode EPR spectrometry. Resonances have been observed at g = 10 and 4.5 in the parallel mode and at g = 10, 5, 1.8 and 1.7 in the normal mode. The bulk of these signals are interpreted to come from a stoichiometric S = 2 system with magnitude of a = 0.17 cm-1, D = +2.1 cm-1, magnitude of E = 0.026 cm-1, g = 2. Exchange coupling between cytochrome a3 and CuB is not indicated.     

Synthesis of well-defined graft copolymers by combination of enzymatic polycondensation and "click" chemistry

Aliphatic polyesters having pendant azide groups were prepared by enzymatic polycondensation in the presence of lipase from Candida antarctica type B (CAL-B). The grafting reaction to the N(3)-functional polyester was carried out quantitatively at room temperature using copper-catalyzed azide-alkyne cycloaddition (CuAAC, "click" reaction) with monoalkyne-functional poly(ethylene oxide) (alkyne-PEO, M(n) = 750 g/mol). Furthermore, both enzymatic polycondensation and "click" reaction were carried out successfully in sequential one-pot reaction. The graft copolymer was surface-active and self-assembled in water. The graft copolymer had a critical aggregation concentration (cac) of 3 × 10(-2) μM in water determined by surface tension measurements. Above cac, the graft copolymer formed single chains and aggregates having a hydrodynamic radius of ～75 nm. Furthermore, the surface activity of the polymers at the air-water interface was studied by Langmuir trough measurements. The Langmuir isotherm of the graft polymer showed a pseudoplateau resulting from desorption of PEO chains into the subphase upon compression.     

Development of a semi-closed underwater breathing apparatus, the "eOBA"

Nippon Sanso K.K developed a compact semi-closed underwater breathing apparatus, the eOBA. It consists of a mouthpiece, manifold with a purge valve, two spring-loaded flexible tubes, a small CO2 absorbent canister (net wt. = 190g), and two compact high pressure bottles (50ccx2: 190kg/cm2: 80%O2, 20%N2) with a regulator which supplies the gas at the constant flow rate of 1.5 l/min and lasts for 10 min. Thus, a counterlung is not incorporated. However, spring-loaded tubes act as a counterlung since its volume increases to 3.5 l when fully inflated. Dives to a depth of 5m are also recommended because of no bypass valve. This new eOBA was tested using the mechanical breathing machine and CO2 supply system to the circuit. For the various combinations of tidal volumes (0.5-2.5 l) and respiratory rates (10-20 breaths/min), the pressure at the mouthpiece, respiratory volume and the CO2 level were continuously monitored. The CO2 absorption rates were then calculated. The thin sloping P-V loops demonstrate that the eOBA is a flow dependent type of apparatus. It was found that the external work of breathing (0.1 kg.m/l at 30 l/min) were allowable. The CO2 absorption rates were sufficient when minute ventilation increased to 30 l/min. Thus, results show that the eOBA must be suitable for shallow and short dives.     

Effect of selected "desirable difficulty" learning strategies on the retention of physiology information

The purpose of this study was to examine the effects of interleaving and expanding retrieval on the retention of physiology concepts. Participants (n = 189) read and then reread 30 immunology and reproductive physiology passages. Half of the participants read and then reread the passages in a blocked manner (e.g., a(1)a(2)a(3)b(1)b(2)b(3)), and the other half did so in an interleaved manner (e.g., a(1)b(1)b(2)a(2)a(3)b(3)). Participants were then repeatedly assessed, without feedback, after either a uniform or an expanding series of intervals. Half of the students from both the blocked and interleaved groups completed the assessments 1, 2, and 3 days after rereading the passages (uniform), whereas the other half completed the assessments immediately and 1 and 3 days after rereading the passages (expanding). All participants completed a final assessment 10 days after rereading the passages. There were no significant differences between the blocked and interleaved groups on any of the assessments, nor were there any significant interactions between the groups on any of the assessments. Those in the expanding retrieval group scored significantly higher than those in the uniform group on all four assessments (ANOVA; assessment 1: F = 35.12, P = 0.00; assessment 2: F = 13.88, P = 0.00; assessment 3: F = 10.87, P = 0.00; and assessment 4: F = 6.79, P = 0.01). Mean final assessment scores were 47.58 ± 19.81 and 40.50 ± 17.17 for the expanding and uniform groups, respectively. The results indicate that participants benefited more from expanding retrieval practice.     

Virulence of Vibrio harveyi responsible for the "Bright-red" Syndrome in the Pacific white shrimp Litopenaeus vannamei

Vibrio harveyi (Vh) CAIM 1792 strain was isolated from Litopenaeus vannamei affected with "Bright-red" Syndrome (BRS). The strain grew in 1-10% NaCl, at 15-35°C and was resistant to ampicillin (10 μg), carbenicillin (100 μg) and oxytetracycline (30 μg). The lowest MIC was for enrofloxacine (0.5 μgml(-1)). The in vivo and in vitro toxicity of bacterial cells and the extracellular products (ECPs) of Vh CAIM 1792 grown at 1.0%, 2.0% and 4.0% NaCl were evaluated. Adherence ability, enzymatic activities and siderophore production of bacterial cell was tested. The ECPs exhibited several enzymatic activities, such as gelatinase, amylase, lipase, phospholipase and caseinase. These ECPs displayed a strong cytotoxic effect on HELA cell line at 6 and 24 h. Challenges using 10(3) CFU g(-1) caused opacity at the site of injection and over 80% shrimp mortality before 24 h p.i. (post-injection). Mortality caused by the ECPs was higher than mortalities with bacteria, especially in the first hours p.i. Bacteria were re-isolated from hemolymph samples of moribund shrimp and identified as Vh CAIM 1792 by rep-PCR. Histological analysis of shrimp L. vannamei injected with Vh CAIM 1792 revealed generalized necrosis involving skeletal muscle (MU) at the injection site, the lymphoid organ (LO), heart and connective tissues. Melanization within the MU at the site of injection was also observed as well as hemocytic nodules within the hearth and MU at 168 h p.i. LO was the target organ of BRS. Necrosis of the MU at the injection site was the main difference in comparison to other shrimp vibriosis.     

Specific metabolic rate, longevity and "Rubner constant" for birds

An assumption was made that age constituent alpha x(beta) of mortality of individuals in a population in Weibull equation mx = m0 + alpha x(beta) (Ricklefs, 2000) reflects change of specific metabolic rate of one individual with age. Based upon that hypothesis a formula was proposed for relationship of specific metabolic rate of an adult individual after cessation of growth, when mass W is attained, and age t: q(t) = q0(1-omega(beta) + 1t(beta)) where q0 = aW(-b) is value q(t) at the moment of growth cessation and omega = alpha(1/(beta + 1)) is "ageing rate", determined and estimated by R. Ricklefs. Maximum longevity of an individual was determined as [equation: see text], where qcrit is specific metabolic rate at the age tmax. Parameter beta and relationships omega(W) and (qcrit/q0)(W) were approximated for birds from data of Ricklefs. Statistical comparison of results of calculations of tmax was carried out on the basis of the above formula and other known formulas for groups of Passeriformes and non-Passeriformes. Rubner constant [equation: see text] was calculated assuming that body mass of an adult individual (W) is attained in the first year of life (tA = 0). Average values of 602.4 +/- 2.5 kcal g(-1) (n = 83) for non-Passeriformes and 963 +/- 6.3 kcal g(-1) (n = 41) for Passeriformes were obtained.     

Efficacy of chlorine and heat treatment in killing Salmonella stanley inoculated onto alfalfa seeds and growth and survival of the pathogen during sprouting and storage.

The efficacy of chlorine and hot water treatments in killing Salmonella stanley inoculated onto alfalfa seeds was determined. Treatment of seeds containing 10(2) to 10(3) CFU/g in 100-micrograms/ml active chlorine solution for 5 or 10 min caused a significant (P < or = 0.05) reduction in population, and treatment in 290-micrograms/ml chlorine solution resulted in a significant reduction compared with treatment in 100 micrograms of chlorine per ml. However, concentrations of chlorine of up to 1,010 micrograms/ml failed to result in further significant reductions. Treatment of seeds containing 10(1) to 10(2) CFU of S. stanley per g for 5 min in a solution containing 2,040 micrograms of chlorine per ml reduced the population to undetectable levels (< 1 CFU/g). Treatment of seeds in water for 5 or 10 min at 54 degrees C caused a significant reduction in the S. stanley population, and treatment at > or = 57 degrees C reduced populations to < or = 1 CFU/g. However, treatment at > or = 54 degrees C for 10 min caused a substantial reduction in viability of the seeds. Treatment at 57 or 60 degrees C for 5 min appears to be effective in killing S. stanley without substantially decreasing germinability of seeds. Storage of seeds for 8 to 9 weeks at 8 and 21 degrees C resulted in reductions in populations of S. stanley of about 1 log10 and 2 log10 CFU/g, respectively. The behavior of S. stanley on seeds during soaking germination, sprouting, and refrigerated storage of sprouts was determined. An initial population of 3.29 log10 CFU/g increased slightly during 6 h of soaking, by about 10(3) CFU/g during a 24-h germination period, and by an additional 10 CFU/g during a 72-h sprouting stage. A population of 10(7) CFU/g of mature alfalfa sprouts was detected throughout a subsequent 10-day storage period at 5 degrees C. These studies indicate that while populations of S. stanley can be greatly reduced, elimination of this organism from alfalfa seeds may not be reliably achieved with traditional disinfection procedures. If S. stanley is present on seeds at the initiation of the sprout production process, populations exceeding 10(7) CFU/g can develop and survive on mature sprouts exposed to handling practices used in commercial production and marketing.     

Improved production of biosurfactant by a <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> mutant using vegetable oil refinery wastes

Biosurfactant production by Pseudomonas aeruginosa EBN-8 mutant was studied in shake flasks on separate wastes from canola, soybean and corn oil refineries. Of the substrates tested, canola oil refinery waste (COD=20 g l⁻¹) supplemented with sodium nitrate (at COD/N=20) showed the best microbial growth (4.50 g l⁻¹) and rhamnolipid production (8.50 g l⁻¹), at 10 d of incubation with the specific growth rate of 0.316 h⁻¹ and specific product yield of 0.597 g g⁻¹ h. Its cell-free supernatant showed the critical micelle dilution (CMD) of 150 and surface tension (ST) of 28.5 mN m⁻¹.     

A "virtual gland" method for quantifying epithelial fluid secretion

We developed a new apparatus, the virtual gland (VG), for measuring the rate of fluid secretion (Jv), its composition, and the transepithelial potential (TEP) in cultured epithelial cells under open circuit. The VG creates a 10-microl chamber above the apical surface of epithelial cells on a Costar filter with a small hole leading to an oil-filled reservoir. After the chamber is primed with a fluid of choice, secreted fluid is forced through the hole into the oil, where it forms a bubble that is monitored optically to determine Jv and collected for analysis. Calu-3 cells were mounted in the VG with a basolateral bath consisting of Krebs-Ringer bicarbonate buffer at 37 degrees C. Basal Jv was 2.7 +/- 0.1 microl x cm(-2) x h(-1) (n = 42), and TEP was -9.2 +/- 0.6 mV (n = 33); both measures were reduced to zero by ouabain (n = 6) x Jv and TEP were stimulated 64 and 59%, respectively, by 5 microM forskolin (n = 10), 173 and 101% by 1 mM 1-ethyl-2-benzimidazolinone (n = 5), 213 and 122% by 333 nM thapsigargin (n = 5), and 520 and 240% by forskolin + thapsigargin (n = 6). Basal Jv and TEP were inhibited to 82 and 63%, respectively, with 10 microM bumetanide (n = 5), 71 and 82% with 100 microM acetazolamide (n = 5), and 47 and 56% with 600 microM glibenclamide (n = 4). Basal Jv and TEP were 52 and 89% of control values, respectively, after HCO3- replacement with HEPES (n = 16). The net HCO3- concentration of the secreted fluid was close to that of the bath (25 mM), except when stimulated with forskolin or VIP, when it increased (approximately 80 mM). These results validate the use of the VG apparatus and provide the first direct measures of Jv in Calu-3 cells.     

Respiration and lactate production in isolated frog skeletal muscle: effects of passive stretch

The effects of varying degrees of passive stretch on in vitro oxygen consumption and intracellular lactate efflux have been investigated in paired recti abdomini muscles from small male frogs. Oxygen consumption [mm3 (STP)/mg (dry wt)/hr] was found to be linearly related to load (r = 0.98), increasing from 1.57 +/- 0.11 (SE) at 2 g to 2.30 +/- 0.18 at 10 g, 2.89 +/- 0.16 at 20 g and 3.26 +/- 0.21 at 30 g. Lactate released into the medium [microM/g (dry wt)/hr] was inversely related to load (r = -0.52), increasing initially from 36.84 +/- 3.28 (SE) at 2 g to 108.55 +/- 12.9 at 10 g, then abruptly decreasing with additional loading (18.10 +/- 2.60 at 20 g and 11.71 +/- 2.80 at 30 g). Results suggest that as stretch-related oxidative energy metabolism increases, there is a lessening dependence on anaerobic energy-yielding processes.     

Overview on coenzyme Q10 as adjunctive therapy in chronic heart failure. Rationale, design and end-points of "Q-symbio"--a multinational trial

Energy starvation of the myocardium is probably a dominant feature of heart failure and attention has been directed towards agents which may stabilize myocardial metabolism and maintain adequate energy stores. A reduced myocardial tissue content of the essential redox-component and natural antioxidant Coenzyme Q10 (CoQ10) has been detected in patients with heart failure and the observed level of CoQ10 deficiency was correlated to the severity of heart failure. CoQ10 fulfills various criteria of an obvious adjunct in patients with symptomatic heart failure: it is devoid of significant side effects and it improves symptoms and quality of life. Till this date, several double-blind placebo-controlled trials with CoQ10 supplementation in more than 1000 patients have been positive and statistically significant with respect to various clinical parameters, e.g. improvement in NYHA Class, exercise capacity and reduced hospitalisation frequency. Also treatment with CoQ10 led to a significant improvement of relevant hemodynamic parameters. In only 3 out of 13 double-blind studies comprising 10% of the total number of patients treated the results were neutral. Thus, based on the available controlled data CoQ10 is a promising, effective and safe approach in chronic heart failure. This is why a double-blind multicenter trial with focus on morbidity and mortality has been planned to start in 2003: Q-SYMBIO. Patients in NYHA classes III to IV (N=550) receiving standard therapy are being randomized to treatment with CoQ10 100 mg t.i.d. or placebo in parallel groups. End-points in a short-term evaluation phase of 3 months include symptoms, functional capacity and biomarker status (BNP). The aim of a subsequent 2-year follow-up study is to test the hypothesis that CoQ10 may reduce cardiovascular morbidity (unplanned cardiovascular hospitalisation due to worsening heart failure) and mortality as a composite endpoint. This trial should help to establish the future role of CoQ10 as part of a maintenance therapy in patients with chronic heart failure.     

HPLC测定葛花中6″-O-木糖鸢尾苷的含量

本文建立了葛花中6″-O-木糖鸢尾苷含量测定的HPLC方法。采用GRACE C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-水梯度洗脱,流速为0.8 mL/min,紫外检测波长为265 nm,柱温为室温。6″-O-木糖鸢尾苷的峰面积(Y)与浓度(X)在10.33～185.99μg/mL范围内线性关系良好,Y=30731X–216635,r=0.9992(n=7);平均回收率为100.2%,RSD<2%(n=9);测得8批葛花药材中6″-O-木糖鸢尾苷含量为11.08～48.23 mg/g。方法学验证结果表明该法准确、可靠,可用于葛花中主要成分6″-O-木糖鸢尾苷的含量测定。     

EPR signals of cytochrome b558 purified from porcine neutrophils.

Cytochrome b558 of pig blood neutrophils was partially purified, and its EPR spectra were measured. The cytochrome b558 was solubilized from membranes with the detergent n-heptyl-beta-thioglucoside and purified by DEAE-Sepharose and heparin-Sepharose chromatographies. The small and large subunits of cytochrome b558 were detected on gel by immunoblotting. A solution of the purified, undenatured cytochrome b558 at 85-108 microM concentration was obtained. The concentrated cytochrome b558 showed an EPR signal at a g value of 3.26 with a bandwidth of 100 G at 10 K. Addition of 2 mM KCN had no effect on the low spin signal at g = 3.26 but caused disappearance of a minor high spin signal. The cyanide-insensitive signal at g = 3.26 disappeared completely on reduction with Na2S2O4. These results suggest that the g = 3.26 signal is characteristic of the low spin heme in cytochrome b558 of neutrophils.     

Synthesis of bacteriochlorophyll <Emphasis Type="Italic">a</Emphasis> by the purple nonsulfur bacterium <Emphasis Type="Italic">Rhodobacter capsulatus</Emphasis>

The ability of purple nonsulfur bacteria Rhodobacter capsulatus B10 to synthesize bacteriochlorophyll under phototrophic and dark conditions was studied. The modes for cultivation in the dark with oxygen limitation in a continuous culture at D = 0.1 h^?1 were selected. The yield of biomass reached 20 g/l; the bacteriochlorophyll a output of the process amounted to 16.6 mg/l h^?1.