Robustness of Sequential Tests for Means

Three sequential tests of the null hypothesis, μ = μ₀, versus an alternative of the form (μ ? μ₀)²= =σ²d² are compared for different forms of violation of the underlying normal assumption. 10000 samples were simulated for d=0.6; 1.0 and 1.6 (σ² = 1), four (α, β)-combinations and seven alternative distributions. The results show that for small d-values one test is robust for α and another for β. For large d all tests can be used.     

Optimisation of reaction conditions for production ofS-(−)-2-hydroxypropiophenone byAcinetobacter calcoaceticus

Summary Whole cells and cell-free extracts ofAcinetobacter calcoaceticus containing benzoylformate decarboxylase efficiently condensed benzoylformate and acetaldehyde to produce the acyloin compoundS-(–)-2-hydroxypropiophenone. Optimal concentrations of acetaldehyde cosubstrate for this reaction were found to be 1600 and 800 mM when whole cells and cell-free extracts were used respectively as biocatalysts. In both cases, optimal benzoylformate concentration was 100 mM. Temperature and pH optima for the biotransformation reaction were 30°C and 6.0 respectively. Under optimised conditions, maximum production of 2-hydroxypropiophenone, amounting to 8.4 g L^–1, occurred after a 2-h incubation. Product formation equivalent to 6.95 g in 1 h corresponded to a productivity of 267 mg acyloin per g dry cells per h.     

Sperm concentration at freezing affects post-thaw quality and fertility of ram semen

We have investigated the effect of sperm concentration in the freezing doses 200, 400, 800, and 1600 × 10⁶ mL⁻¹ on the post-thaw quality and fertility of ram semen. Semen was collected from seven adult Churra rams by artificial vagina during the breeding season. The semen was diluted in an extender (TES-Tris-fructose, 20% egg yolk, and 4% glycerol), to a final concentration of 200, 400, 800, or 1600 × 10⁶ mL⁻¹ and frozen. Doses were analyzed post-thawing for motility (computer-assisted sperm analysis system [CASA]), viability, and acrosomal status (fluorescence probes propidium iodide [PI]/peanut agglutinin conjugated with fluorescein thiocyanate (PNA-FITC), SYBR-14/PI [Invitrogen; Barcelona, Spain] and YO-PRO-1/PI [Invitrogen; Barcelona, Spain]). Total motility and velocity were lower for 1600 × 10⁶ mL⁻¹ doses, while progressive motility and viability were lower both for 800 and 1600 × 10⁶ mL⁻¹. The proportion of viable spermatozoa showing increased membrane permeability (YO-PRO-1+) rose in 800 and 1200 × 10⁶ mL⁻¹. Intrauterine inseminations were performed with the 200, 400, and 800 × 10⁶ mL⁻¹ doses at a fixed sperm number (25 × 10⁶ per uterine horn) in synchronized ewes. Fertility (lambing rate) was similar for semen frozen at 200 (57.5%) or 400 × 10⁶ mL⁻¹ (54.4%), whereas it was significantly lower for 800 × 10⁶ mL⁻¹ (45.5%). In conclusion, increasing sperm concentration in cryopreserved semen, at least at 800 × 10⁶ mL⁻¹ and more, adversely affects the postthawing quality and fertility of ram semen.     

Unconditional Tests of Hypotheses Concerning Relative Risk

The problem of estimation of, and statistical tests for, the relative risk (=α) from samples in which there is matching on covariates have been discussed by various authors (Cox, 1970; MANTEL and HAENSZEL, 1959) using the approach of conditional variates. This paper discusses log LR (likelihood ratio) tests of hypotheses concerning α, as based on one or more sets of matched samples. Approximate X² tests are also developed for the hypotheses concerning α. An example is presented which illustrates the proposed tests of significance (Table 1).     

The effect of different levels of the B800-850 light-harvesting complex on intracytoplasmic membrane development in Rhodobacter sphaeroides

A role for the peripheral (B800-850) light-harvesting complex in vesicularization of the Rhodobacter sphaeroides intracytoplasmic membrane (ICM), suggested from studies in mutant strains lacking one or more of the pigment-protein complexes, was examined further in the wild-type strain NCIB 8253 grown at high (∼1000 W m^–2), moderate (∼300 W m^–2), and low (∼100 W m^–2) light intensities. The resulting ICM vesicles (chromatophores) had B800-850 levels related inversely to irradiance and banded in rate-zone sedimentation at ∼1.10, 1.09, and 1.07 g ml^–1, respectively. Equilibrium centrifugation on iso-osmotic gradients indicated that this distinct sedimentation behavior resulted solely from differences in hydrodynamic radii. These size differences were confirmed by gel-exclusion chromatography and in electron micrographs of thin-sectioned cells. A pulse-chase study of ICM growth following a tenfold reduction in light intensity showed a relatively slow equilibration of membrane proteins during adaptation, and that new protein was incorporated largely into additional ICM formed at the lowered illumination level, giving rise to chromatophores of reduced size and elevated B800-850 content. These results provide further evidence for a model in which the B800-850 complex both drives development of vesicular ICM in Rba. sphaeroides and determines the size of resulting vesicles. Received: 12 October 1995 / Accepted: 21 December 1995     

The Purification and Characterization of Gemini Particles from Abutilon Mosaic Virus Infected Malvaceae

Gemini particles were purified from Abutilon Mosaic Virus infected Abutilon sellovianum, Sida micrantha and Malva parviflora. They sediment with 82 S, possess a density of 1.30 g/cm³ in Cs₂SO₄ and a nucleic acid percentage of 18 % calculated from ultra-violett spectrometry. Their proteins are separated in SDS-PAGE into two components of 28 000 and 27 000 mol. wt. Their nucleic acid consists of single-stranded circular DNA with length distribution maxima of 2 800, 2 500 and 1 300 bases for the circular molecules and 2 050, 1 850, and 1 300 bases for the correspondmg linear molecules.     

Growth and ion uptake in <Emphasis Type="Italic">Annona muricata</Emphasis> and <Emphasis Type="Italic">A. squamosa</Emphasis> subjected to salt stress

The effects of treatment with NaCl (3, 100 and 300 mM) for 1, 2, 3 and 7 d on plant growth and ion accumulation were analyzed in 2-week and 8-week-old Annona muricata and A. squamosa plants. Fresh mass and root growth inhibition were directly related to the increase in salinity, particularly for A. squamosa. Two-weeks old seedlings were sensitive to 100 and 300 mM NaCl particularly after 7 d, whereas 8-week-old plants were shown to be more resistant to NaCl even at 300 mM NaCl. Na⁺ and Cl^– mostly accumulated in young leaves. Our results suggest that A. squamosa is more sensitive than A. muricata to salt stress and that older seedlings of both species are more tolerant than younger seedlings.     

Changes in carbon storage and fluxes in a chronosequence of ponderosa pine

Forest development following stand‐replacing disturbance influences a variety of ecosystem processes including carbon exchange with the atmosphere. On a series of ponderosa pine (Pinius ponderosa var. Laws.) stands ranging from 9 to> 300 years in central Oregon, USA, we used biological measurements to estimate carbon storage in vegetation and soil pools, net primary productivity (NPP) and net ecosystem productivity (NEP) to examine variation with stand age. Measurements were made on plots representing four age classes with three replications: initiation (I, 9–23 years), young (Y, 56–89 years), mature (M, 95–106 years), and old (O, 190–316 years) stands typical of the forest type in the region. Net ecosystem productivity was lowest in the I stands (?124 g C m^?2 yr^?1), moderate in Y stands (118 g C m^?2 yr^?1), highest in M stands (170 g C m^?2 yr^?1), and low in the O stands (35 g C m^?2 yr^?1). Net primary productivity followed similar trends, but did not decline as much in the O stands. The ratio of fine root to foliage carbon was highest in the I stands, which is likely necessary for establishment in the semiarid environment, where forests are subject to drought during the growing season (300–800 mm precipitation per year). Carbon storage in live mass was the highest in the O stands (mean 17.6 kg C m^?2). Total ecosystem carbon storage and the fraction of ecosystem carbon in aboveground wood mass increased rapidly until 150–200 years, and did not decline in older stands. Forest inventory data on 950 ponderosa pine plots in Oregon show that the greatest proportion of plots exist in stands ～ 100 years old, indicating that a majority of stands are approaching maximum carbon storage and net carbon uptake. Our data suggests that NEP averages ～ 70 g C m^?2 year^?1 for ponderosa pine forests in Oregon. About 85% of the total carbon storage in biomass on the survey plots exists in stands greater than 100 years, which has implications for managing forests for carbon sequestration. To investigate variation in carbon storage and fluxes with disturbance, simulation with process models requires a dynamic parameterization for biomass allocation that depends on stand age, and should include a representation of competition between multiple plant functional types for space, water, and nutrients.     

Efficiency in the Use of Hotellings T2

Alternative scale models are compared with regard to the Pitman efficiency of Hotelling's T² in tests for means. A basic decomposition identifies subspaces of the parameters in which one model is more efficient than another, or the two models have equal efficiency. Local and global bounds for relative Pitman efficiencies are given with reference to these subspaces. Applications include efficiency comparisons of marginal and conditional tests in a given model, and a comparison of the sensitivities of selected T² tests regarding psychological profiles in treated and control groups of children.     

Effects of Straw Return in Deep Soils with Urea Addition on the Soil Organic Carbon Fractions in a Semi-Arid Temperate Cornfield

Returning straw to deep soil layers by using a deep-ditching-ridge-ploughing method is an innovative management practice that improves soil quality by increasing the soil organic carbon (SOC) content. However, the optimum quantity of straw return has not been determined. To solve this practical production problem, the following treatments with different amounts of corn straw were investigated: no straw return, CK; 400 kg ha^-1 straw, S₄₀₀; 800 kg ha^-1 straw, S₈₀₀; 1200 kg ha^-1 straw, S₁₂₀₀; and 1600 kg ha^-1 straw, S₁₆₀₀. After straw was returned to the soil for two years, the microbial biomass C (MBC), easily oxidized organic C (EOC), dissolved organic C (DOC) and light fraction organic C (LFOC) content were measured at three soil depths (0–10, 10–20, and 20–40 cm). The results showed that the combined application of 800 kg ha^-1 straw significantly increased the EOC, MBC, and LFOC contents and was a suitable agricultural practice for this region. Moreover, our results demonstrated that returning straw to deep soil layers was effective for increasing the SOC content.     

Interactive effects of soil salinity and boron on growth,mineral composition and CO<Subscript>2</Subscript> assimilation of pistachio seedlings

Pistachio is a tree of the arid and semi-arid regions where salinity and boron (B) toxicity can be major environmental stresses. In this study, individual and combined effects of different concentrations of NaCl (0, 800, 1600, 2400 and 3200 mg kg^?1 soil) and B (0, 2.5, 5.0, 10.0 and 20.0 mg kg^?1 soil) were studied on growth, gas-exchange and mineral composition of pistachio seedlings for a duration of 120 days. Excess amounts of salinity (> 1600 mg NaCl kg^?1 soil) and B (20.0 mg kg^?1 soil) significantly reduced the plant growth and CO₂ assimilation, which was associated with accumulation of Na, Cl and B in leaves. There was also a decline in cell membrane stability index (MSI). Reduced stomatal conductance (g _s) was the primary cause of inhibition of photosynthesis rate (P _N) under mild to moderate salinity. However, under severe salt stress and B toxicity, non-stomatal effects contributed to the inhibition of CO₂ assimilation in addition to the decline in g _s value. Application of 5.0–10.0 mg B kg^?1 soil significantly improved the plant growth and P _N and also recovered the MSI as countermeasures against salt stress. These observations were related to the role of B in cell membrane structure and functioning which reduced the concentration of toxic ions in the leaves. However, cell membrane damages and chlorophyll loss in plants affected by salt were observed to be exacerbated when excess amounts of B were present. In conclusion, the results revealed that optimizing the B nutrition can improve the performance of pistachio seedlings under salt stress, and NaCl also showed a mitigating effect on B toxicity if its concentration in the soil is kept under the plant salt tolerance threshold.     

The flow of water into fruit trees. I. Resistances to water flow through roots and stems

Relative conductivity (K) to water in healthy apple trees ranged from maximum values of 18.2 cm³.100 s^-1.cm length.0.001 Pas.kPa^-1.cm^-2 xylem area, for major suberized roots to values of 1.6 for 1-yr-old twigs. The values for equivalent parts of healthy cherry trees were 26.3 and 3.3. Trees with roots affected by the larvae of the fruit tree root weevil (Leptopius squalidus) which causes either chronic growth decline or sudden wilting and death, had values as low as 1% of healthy trees, in those parts of the tree showing wilting and lack of growth. Water flow under pressure into the root systems of healthy apple trees increased linearly with increases in pressure from 200 to 800 kPa. Flows into dormant and active root systems respectively were 0.6 and 1.7 cm³.100 s^-1. 100 cm² root surface area. 100 kPa^-1.     

Genotoxic and Cytotoxic Effects of Antiretroviral Combinations in Mice Bone Marrow

Commonly used guidelines for the management of human immunodeficiency virus (HIV) infection (highly active antiretroviral therapy, HAART) include drug combinations such as tenofovir disoproxil fumarate (TDF) + lamivudine (3TC) and combivir [zidovudine (AZT) + 3TC] + efavirenz (EFV). These combinations may enhance the genotoxic effects induced by such drugs individually, since the therapy requires lifelong adherence and the drugs have unknown effects during treatment. Thus, the evaluation of the benefits and risks of HAART is of great importance. In order to assess the cytotoxic and genotoxic potential of three concentrations of each of the antiretroviral combinations TDF + 3TC (800 + 400, 1600 + 800, and 3200 + 1600 mg/kg body weight, BW) and combivir + EFV (200 + 100 + 400, 400 + 200 + 800, and 800 + 400 + 1600 mg/kg BW) after two exposure periods (24 h and 48 h), in the present study the in vivo comet assay (single-cell gel electrophoresis) and the mouse bone marrow micronucleus test were used. Neither TDF + 3TC nor combivir + EFV induced DNA damage at any concentrations tested after 24 h or 48 h using the comet assay. After 24 h, both combinations increased the micronucleus frequency at all concentrations tested. After 48 h, combivir + EFV increased the micronucleated polychromatic erythrocyte (MNPCE) frequency at the two highest concentrations tested. Polychromatic erythrocytes (PCE)/normochromatic erythrocytes (NCE) ratio was high for both combinations, suggesting that they can be mitogenic. Since genotoxicity may be related to carcinogenesis, it is necessary to conduct further studies to verify the long-term mutagenic effects of these drugs.     

Insights into the helix‐sense inversion of poly(β‐phenethyl‐L‐aspartate) by two‐dimensional Raman correlation spectroscopy

In this study, the transition process of the helix‐sense inversion of poly(β‐phenethyl‐L‐aspartate) was investigated by Raman scattering and 2‐dimensional correlation spectroscopy. Temperature‐dependent Raman spectra were obtained during the helix‐sense inversion. The results of 2‐dimensional correlation analysis in the spectral regions of 1600‐1800 and 3200‐3400 cm^?1 showed that the intensity changes of the side‐chain ester C═O stretching bands occurred prior to those of amide A and amide I bands in the unwinding process of αR‐helix on heating. The sequential order of the intensity changes for amide A, amide I, and the side‐chain ester C═O stretching bands during the inversion process was determined. It was found that the conformation change of the side chain occurred prior to that of the main chain for the αR‐helix on heating. Thus, we concluded that the transformation of the backbone chain from right‐handed to left‐handed is triggered by the conformational change of the side chains.     

The effect of phosphate rock dissolution on soil chemical properties and wheat seedling root elongation

Soils of the Appalachian region of the United States are acidic and deficient in P. North Carolina phosphate rock (PR), a highly substituted fluoroapatite, should be quite reactive in these soils, allowing it to serve both as a source of P and a potential ameliorant of soil acidity. An experiment was conducted to evaluate the influence of PR dissolution on soil chemical properties and wheat (Triticum aestivum cv. Hart) seedling root elongation. Ten treatments including nine rates of PR (0, 12.5, 25, 50, 100, 200, 400, 800, and 1600 mg P kg^-1) and a CaCO₃ (1000 mg kg^-1) control were mixed with two acidic soils, moistened to a level corresponding to 33 kPa moisture tension and incubated for 30 days. Pregerminated wheat seedlings were grown for three days in the PR treated soils and the CaCO₃ control. Root length was significantly (P<0.05) increased both by PR treatments and CaCO₃, indicating that PR dissolution was ameliorating soil acidity. The PR treatments increased soil pH, exchangeable Ca, and soil solution Ca while lowering exchangeable Al and 0.01 M CaCl₂ extractable soil Al. Root growth in PR treatments was best described by an exponential equation (P<0.01) containing 0.01 M CaCl₂ extractable Al. The PR dissolution did not reduce total soil solution Al, but did release Al complexing anions into soil solution, which along with increased pH, shifted Al speciation from toxic to nontoxic forms. These results suggest that North Carolina PR should contribute to amelioration of soil acidity in acidic, low CEC soils of the Appalachian region.     

The relationship between dose of vitamin E and suppression of oxidative stress in humans

The oxidation hypothesis of atherogenesis has been the focus of much research over the past 2 decades. However, randomized placebo-controlled trials evaluating the efficacy of vitamin E in preventing cardiovascular events in aggregate have failed to show a beneficial effect. Implicit in these trials is that the dose of vitamin E tested effectively suppressed oxidative stress status but this was never determined. We defined the dose-dependent effects of vitamin E (RRR-alpha-tocopherol) to suppress plasma concentrations of F2-isoprostanes, a biomarker of free radical-mediated lipid peroxidation, in participants with polygenic hypercholesterolemia and enhanced oxidative stress, a population at risk for cardiovascular events. A time-course study was first performed in participants supplemented with 3200 IU/day of vitamin E for 20 weeks. A dose-ranging study was then performed in participants supplemented with 0, 100, 200, 400, 800, 1600, or 3200 IU/day of vitamin E for 16 weeks. In the time-course study, maximum suppression of plasma F2-isoprostane concentrations did not occur until 16 weeks of supplementation. In the dose-ranging study there was a linear trend between the dosage of vitamin E and percentage reduction in plasma F2-isoprostane concentrations which reached significance at doses of 1600 IU (35+/-2%, p<0.035) and 3200 IU (49+/-10%, p<0.005). This study provides information on the dosage of vitamin E that decreases systemic oxidant stress in vivo in humans and informs the planning and evaluation of clinical studies that assess the efficacy of vitamin E to mitigate disease.     

Rapid, reversible alterations in spinach thylakoid appression upon changes in light intensity

Changes in light quantity and quality cause structural changes within the thylakoid membrane; long‐term responses have been described for so‐called ‘sun’ and ‘shade’ leaves. Many leaves, however, experience changes in irradiance on a time scale of minutes due to self‐shading and sun flecks. In this study, mature, attached spinach leaves were grown at 300 µmol photons m^?2 s^?1 then rapidly switched to a different light treatment. The treatment irradiances were 10, 800 or 1500 µmol m^?2 s^?1 for 10 min, or 10 or 20 min of self‐shading (about 10 µmol m^?2 s^?1). Image analysis of transmission electron micrographs revealed that a 10 min switch to a lower light intensity increased grana size and number per chloroplast profile by 10–20%. Returning the leaves to 300 µmol m^?2 s^?1 for 10 min reversed the phenomenon. Chlorophyll fluorescence measurements of detached, intact leaves at 77 K were suggestive of a transition from state 2 to state 1 upon shading. Diurnal ultrastructural measurements of granal size and number did not reveal a significant net change in ultrastructure over the time scale of hours. It is concluded that spinach chloroplasts can alter the degree of thylakoid appression in response to irradiance changes on a time scale of minutes. These ultrastructural responses are caused by biochemical and biophysical adjustments within the thylakoid membrane that serve to maximize photosynthesis and minimize photo‐inhibition under rapidly fluctuating light environments.     

Damage functions and thermal requirements of Meloidogyne javanica and Meloidogyne incognita on watermelon

The relationship between the initial (P_i) and final (P_f) population densities of Meloidogyne javanica and yield of watermelon, Citrullus lanatus, cv. Sugar Baby were determined in pot and field experiments. In the pots, the maximum reproduction rate of the nematode was 14, and the equilibrium density was 49 400 eggs/100 cm³ of soil. Yield data represented as fresh top weight fitted the Seinhorst damage function (P < 0.001, R² = 0.7), and the minimum relative yield (m) was 0.65 at P_i ≥ 3200 eggs/100 cm³ of soil and the tolerance limit (T) 74 eggs/100 cm³. In the field experiments (2011 and 2012), the maximum reproduction rate was 73 and 70, and the equilibrium density 32 and 35 second‐stage juveniles (J2)/100 cm³ soil. Yield data represented as fruit weight fitted the Seinhorst damage function in 2011 (P < 0.001, R² = 0.92) and the m‐ and T‐values were 0.63 and 20 J2/100 cm³ of soil, respectively. Meloidogyne incognita and M. javanica needed similar length of time for development to egg‐laying females and life cycle completion at 24.4°C.     

Asymptotical Tests on the Equivalence,Substantial Difference and Non‐inferiority Problems with Two Proportions

Let d = p₂ ? p₁ be the difference between two binomial proportions obtained from two independent trials. For parameter d, three pairs of hypothesis may be of interest: H₁: d ≤ δ vs. K₁: d > δ; H₂: d ? (δ₁, δ₂) vs. K₂: d ∈ (δ₁, δ₂); and H₃: d ∈ [δ₁, δ₂] vs. K₃: d ? [δ₁, δ₂], where H_i is the null hypothesis and K_i is the alternative hypothesis. These tests are useful in clinical trials, pharmacological and vaccine studies and in statistics generally. The three problems may be investigated by exact unconditional tests when the sample sizes are moderate. Otherwise, one should use approximate (or asymptotical) tests generally based on a Z‐statistics like those suggested in the paper. The article defines a new procedure for testing H₂ or H₃, demonstrates that this is more powerful than tests based on confidence intervals (the classic TOST – two one sided tests – test), defines two corrections for continuity which reduce the liberality of the three tests, and selects the one that behaves better. The programs for executing the unconditional exact and asymptotic tests described in the paper can be loaded at http://www.ugr.es/~bioest/software.htm. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Tests with Daphnia magna: A new approach to prescreen toxicity of newly synthesized acetylcholinesterase reactivators

Reactivators of phosphorylated acetylcholinesterase (oximes) are substances used as a human antidotal therapy for organophosphate poisoning. The objective of our study was to examine if juveniles of freshwater microcrustacean Daphnia magna could be employed as test animals in early screen toxicity tests of those substances as a first step for further experiments with daphnids intoxicated by organophosphates. For this purpose, seven different oximes were investigated. It was found that toxicity of all tested oximes increased with time. Mono-quaternary oximes were approximately ten fold (EC₅₀, 14.9 mg.l^{? 1}) more toxic in 24 hour tests and five fold (EC₅₀ was 79.46 mg.l^{? 1}) more toxic in 48 hour tests than bis-quaternary oximes. Tests with daphnids were shown to be easy to carry out at low cost and provided valuable results which could be used as a starting point for further research.