Optimum temperature for egg development of phenotypes in Haemonchus contortus cayugensis as determined by Arrhenius diagrams and Sacher's entropy function

Lejambre L. P. and Whitlock J. H. 1973. Optimum temperature for egg development of phenotypes in Haemonchus contortus cayugensis as determined by Arrhenius diagrams and Sacher's entropy function. International Journal for Parasitology3: 299–310. Oxygen uptake, maturation time and percentage hatch were determined on eggs from “wild-type” Haemonchus contortus cayugensis populations as well as those from the morph types smooth, linguiform B and linguiform A. Sacher's organizational entropy, which is essentially a sum of oxygen utilized by a system corrected for the number of viable units in the system, was used to determine the optimum temperature for development of the eggs. Although the eggs from “wild-type” H. contortus cayugensis hatched well across a broad temperature range; individual phenotypes showed a much narrower range. The optimum temperature for the development of eggs from smooth worms was approximately 30°C while linguiform A and B hatched best at 25°C. Linguiform A eggs had a broader temperature tolerance than linguiform B so that at temperatures of 17°C they hatched with a greater efficiency than did either smooth or linguiform B. It is argued that H. contortus follows a “strategy” of having many phenotypes, each one of which appears to be adapted to a different temperature. This would allow a population to maintain a broad range of optimal temperatures without the expense of maintaining the cybernetic machinery which a single individual would have to have if it were to tolerate the same range of temperature.     

Arrhenius plots and the involvement of thermotropic phase transitions of the thylakoid membrane in chilling impairment of photosynthesis in thermophilic higher plants

Abstract Breaks and discontinuities in Arrhenius plots of physiological and physical properties of thylakoids are not diagnostic of thermotropic lipid phase transitions of the membrane. Bulk lipid transitions, as first inferred by the membrane phase transition hypothesis, do not occur in any higher plant at chilling temperatures. Solidification of some varying, but always minor, fraction of the total membrane lipid does take place. However, the presence of minor domains of solid thylakoid membrane lipid at chilling temperatures is not unique to chilling sensitive plants but is also found in tolerant species. Minor solidification may in some plants, or groups of plants, be controlled by the specific molecular species of phosphatidylglycerol only recently investigated. In plants containing little, or no, phosphatidylglycerol with this positional distribution of fatty acids, other yet unknown constituents of the membrane must fill a similar function, since DSC thermograms indicate minor solidification also in isolated, unperturbed thylakoids from chilling tolerant species. However, chilling induced phase transitions, or other perturbations, of the thylakoid membrane are not the reason for the chilling lability of net photosynthesis in the intact plant. This conclusion follows from detailed comparison between photosynthetic membranes isolated from prechilled plants and the effects of chilling exposure on CO₂ fixation of the whole plant. Damage at the level of the thylakoid membrane does occur, although not to the extent where it can account for the proportionally much larger damage to CO₂ fixation.     

High-resolution profiles of temperature and dissolved oxygen in a river

Longitudinal profiles of water quality along a well-mixed tidal river are, ideally, based on simultaneous measurements at fixed stations distributed along the river. The resolution of the profiles is limited by the density of the stations. However, for a given number of stations the resolution is greatly increased if water quality date can be extrapolated upstream and downstream of the stations, making use of velocity data; the resolution is then determined by the density of the extrapolated data points, which may be an order of magnitude higher than the density of the stations. A 15-km length of river was investigated using 5 current meters equipped to measure depth, temperature, conductivity and dissolved oxygen. Data were recorded simultaneously every 10 minutes. When the average cross-sectional speed was 0.25 ms⁻¹ (typical of tidal conditions), the extrapolated data points were 150 m apart, so the resolution of the resulting profiles (7 per kilometre) was much higher than that of the stations alone (0.3 per kilometre). The extrapolation process required a means of deducing the average cross-sectional speed from the speed measured at a given station. The key to this was provided by temperature data recorded during the onset of a spate, when tidal flow was suspended and the average cross-sectional speed was uniformly about 0.75 ms ⁻¹ at four of the stations. Profiles of temperature and dissolved oxygen were generated by this method; the resolution was about 2 data points km⁻¹ during the onset of the spate, and 6 points km⁻¹ during tidal flow.     

Role of proteins and lipids in non-linear Arrhenius plots of Drosophila mitochondrial succinate-cytochrome c reductase studied by rebinding experiments

Abrupt changes in the Arrhenius activation energy of membrane-bound enzymes have often been correlated with changes in the physical state of membrane phospholipids. Similar changes in activation energy have also been found in soluble enzymes. The possibility exists, therefore, that in some of the membrane-bound enzymes the changes might reflect intrinsic changes of the proteins independent of changes in the membrane phospholipids. This hypothesis was investigated using Drosophila mitochondria isolated from wild type and the mutant Ocd^ts-1. In this mutant it has been shown that succinate-cytochrome $\text{c}$ reductase exhibits a change in Arrhenius activation energy at 18°C which is not found in the wild type (Sondergaard, L., Nielsen, N.C. and Smillie, R.M. (1975) FEBS lett. 50, 126–129). A quantitative thin-layer chromatographic analysis of mitochondrial phospholipids showed sphingomyelin to be more abundant in the wild type than in the mutant (5.2% and 4.3% of the total phospholipids, respectively). Since it was shown that the succinate-cytochrome $\text{c}$ reductase had a lipid requirement for full activity, reciprocal rebinding experiments were done. These experiments showed that the reconstituted membranes exhibited the change in activation energy at 18°C only when the protein moiety came from mutant mitochondria, that is, the change was independent of the source of the phospholipids used.     

Interaction of phospholipid dispersions with water-soluble porphyrins as monitored by their Raman temperature profiles

Raman scattering spectra of 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) dispersions, mixed with water-soluble porphyrins, i.e. cationic copper(II)-5,10,15,12-tetrakis(4-N-methylpyridyl) and anionic silver(II)-5,10,15,20-tetrakis(4-carboxyphenyl)porphyrins, were measured in the 2800-3100 cm(-1) C-H stretching vibration region as a function of the temperature within the 5-55 degrees C range. Temperature profiles of Raman data were constructed from a quantitative data treatment based on factor analysis. This method is shown to be more efficient than the commonly used approach employing peak intensity ratios. Parameters of the gel phase to liquid crystal phase transition determined from Raman temperature profiles were used to monitor the porphyrin influence on DPPG and DPPC structures. Both negatively and positively charged porphyrins significantly perturb DPPC and DPPG dispersions, causing significant downshift of the transition temperature and broadening of the transition region. Water-soluble porphyrins are assumed to set at the outside part of phospholipid dispersions and interact via coulombic forces with charged lipid heads. For the cationic CuTMPyP, the strongest effect has been observed for negatively charged DPPG. In contrast, anionic AgTPPC4 has been found to interact more efficiently with DPPC possessing both positive and negative charges.     

A photoacoustic method for rapid assessment of temperature effects on photosynthesis

The photosynthetic and photoacoustic properties of leaf samples were studied using a photoacoustic system modified for precise temperature control. Data were collected over a temperature range of −10 °C to +60 °C. A distinct acoustic noise transient marked the freezing temperature of the samples. A positive absorption transient and a brief period of oxygen uptake marked the thermal denaturing temperature of the samples. Between these extremes, the effects of temperature on light absorption, oxygen evolution, and photochemical energy storage were quantified quickly and easily. Oxygen evolution could be measured as low as −5 °C and showed a broad temperature peak that was 10 °C lower under limiting light intensity than under saturating light intensity. Photochemical energy storage showed a narrower temperature peak that was only slightly lower for limiting light intensities than for saturating light intensities. In a survey of diverse plants, temperature response curves for oxygen evolution were determined readily for a variety of leaf types, including ferns and conifer needles. These results demonstrate that temperature-controlled photoacoustics can be useful for rapid assessment of temperature effects on photosynthesis and other leaf properties.     

Determination of melting temperature and temperature melting range for DNA with multi-peak differential melting curves

Many factors that change the temperature position and interval of the DNA helix–coil transition often also alter the shape of multi-peak differential melting curves (DMCs). For DNAs with a multi-peak DMC, there is no agreement on the most useful definition for the melting temperature, T_m, and temperature melting width, ΔT, of the entire DNA transition. Changes in T_m and ΔT can reflect unstable variation of the shape of the DMC as well as alterations in DNA thermal stability and heterogeneity. Here, experiments and computer modeling for DNA multi-peak DMCs varying under different factors allowed testing of several methods of defining T_m and ΔT. Indeed, some of the methods give unreasonable “jagged” T_m and ΔT dependences on varying relative concentration of DNA chemical modifications (r_b), [Na⁺], and GC content. At the same time, T_m determined as the helix–coil transition average temperature, and ΔT, which is proportional to the average absolute temperature deviation from this temperature, are suitable to characterize multi-peak DMCs. They give smoothly varying theoretical and experimental dependences of T_m and ΔT on r_b, [Na⁺], and GC content. For multi-peak DMCs, T_m value determined in this way is the closest to the thermodynamic melting temperature (the helix–coil transition enthalpy/entropy ratio).     

A rapid radioimmunoassay for determining plasma concentrations of LH in dogs

A heterologous dog LH radioimmunoassay was modified to provide accurate results for LH concentrations in blood plasma of dogs within 3-4 h. This assay utilizes radioiodinated ovine LH (LER-1056-C2), antiserum against ovine LH (GDN-15) at a final dilution of 1:48,000 and dog LH (LER-1685-1) as standard. A 60-min incubation, including a 30-min delay in the addition of tracer, was carried out at 37 degrees C. The free and antibody-bound hormone were separated by addition of a Micro Sepharose bead suspension containing anti-gamma-globulin, followed by incubation at room temperature for 30 min. The minimum detectable concentration in this assay, calculated from the precision profile, was 1.5 micrograms/l. The amount of dog LH needed to cause 50% reduction of the initial binding was 1.57 +/- 0.13 ng/tube (15.7 micrograms/l for 100-microliters samples). Daily blood samples were collected in heparinized tubes from the cephalic vein of 5 pointer and 7 beagle bitches from the onset of pro-oestrus until 3-4 days after either the last mating or artificial insemination with frozen semen or until metoestrus. Samples were assayed for LH content by the short and normal incubations as well as for progesterone and oestradiol-17 beta content. In all bitches plasma concentrations of progesterone increased rapidly within 1 week after the LH peak which indicates that they had ovulated. Comparison of the short (1.5 h) with the normal (24 h) incubation system resulted in a regression equation: y = 1.0 + 0.7 x (r = 0.95, n = 153 samples).(ABSTRACT TRUNCATED AT 250 WORDS)     

A method for rapid freeze fixation of plant cells

Summary We describe here an apparatus that permits rapid freeze fixation of whole cells, which are then prepared by freeze substitution and resin embedment for examination in the EM. The freezing device utilizes a rotary solenoid that rapidly plunges the specimen holder, a formvar-film-covered thin wire loop, into a well of stirred liquid propane at –180C. The rotary solenoid allows for an adjustable, repeatable immersion rate. Substitution takes place at –80 C in acetone with 2% OsO₄ and is followed by en bloc staining in either hafnium tetrachloride or uranyl acetate. We have utilized these techniques on plant cells, for which there has been relatively little published work when compared to other organisms. The results show that, with the versatile specimen holder and rapid, repeatable immersion rates, different cell types, including pollen, stamen hairs, and germinating moss spores, can be rapidly frozen with repeatable success. The improved preservation achieved with rapid freeze fixation over conventional chemical fixation reveals itself particularly in the structure of the plasmamembrane, the cytoskeleton, chromatin, and certain endomembrane systems.     

A review of bioluminescent ATP techniques in rapid microbiology

Use of firefly luciferase to assay adenosine triphosphate (ATP) extracted from microorganisms provides an easy means to enumerate microbes within minutes. The small amount of light produced is proportional to ATP and thus microbial number. The average bacterium contains around 10^?15 g ATP per cell. Present reagents permit detection of 10³ cells per tube. Luminometers currently on the market detect about 10^?12 g ATP. Proper extraction of ATP from the microbes is an essential part of any protocol, as is the removal of non-microbial ATP from, for example, somatic cells also present in samples. The technique may be applied to a wide range of samples, for example food and beverages and clinical samples such as urine. The ATP assay gives a global measure of microbial numbers, i.e. it is not species specific unless a species separation step is included in the protocol.     

A graphical model for the evaluation of cross-transfer evidence in DNA profiles

The role of graphical models in the assessment of transfer evidence is described with particular reference to the role of cross-transfer evidence. The issues involved in the determination of factors (nodes), associations (links) and probabilities to be included are discussed. Four types of subjective probabilities are of particular interest: those for transfer, persistence and recovery; innocent acquisition; relevance; innocent presence. Examples are given to illustrate the roles of various aspects of the suspect's and victim's lifestyle and the investigation of the evidence found on the suspect and victim in assessing the probability of ultimate issue, that the suspect committed the crime.     

Development of a self-contained,indwelling vaginal temperature probe for use in cattle research

A device was developed to monitor the vaginal temperature of cattle in a research setting. This device decreases labor involved with monitoring body temperature compared with manual temperature readings, allows for continuous monitoring of vaginal temperature at 1 min intervals, and also allows for temperature measurements without the presence of a human handler or without restraint, which can agitate cattle. The device consists of a blank controlled internal drug release (CIDR) device (designed by Pfizer Animal Health as an indwelling vaginal probe) that holds an indwelling vaginal temperature probe logger. The fabrication of the vaginal probe costs approximately US $325 per unit. Similar rectal and vaginal temperature responses to lipopolysaccharide challenge were observed when vaginal and rectal temperatures were measured simultaneously in the same heifer (P>0.05). Additionally, rectal and vaginal temperatures were highly correlated (r=0.97; P<0.0001). Similar to the rectal temperature monitoring device, the vaginal device allows for the measurement of vaginal temperature without the potential biases associated with the stress response produced as a reaction to the handling by and (or) presence of humans. The vaginal temperature recording device will provide researchers with an additional inexpensive tool to study physiological responses in female cattle.     

A rapid decrease in temperature induces latewood formation in artificially reactivated cambium of conifer stems

Background and Aims Latewood formation in conifers occurs during the later part of the growing season, when the cell division activity of the cambium declines. Changes in temperature might be important for wood formation in trees. Therefore, the effects of a rapid decrease in temperature on cellular morphology of tracheids were investigated in localized heating-induced cambial reactivation in Cryptomeria japonica trees and in Abies firma seedlings. Methods Electric heating tape and heating ribbon were wrapped on the stems of C. japonica trees and A. firma seedlings. Heating was discontinued when 11 or 12 and eight or nine radial files of differentiating and differentiated tracheids had been produced in C. japonica and A. firma stems, respectively. Tracheid diameter, cell wall thickness, percentage of cell wall area and percentage of lumen area were determined by image analysis of transverse sections and scanning electron microscopy. Key Results Localized heating induced earlier cambial reactivation and xylem differentiation in stems of C. japonica and A. firma as compared with non-heated stems. One week after cessation of heating, there were no obvious changes in the dimensions of the differentiating tracheids in the samples from adult C. japonica. In contrast, tracheids with a smaller diameter were observed in A. firma seedlings after 1 week of cessation of heating. Two or three weeks after cessation of heating, tracheids with reduced diameters and thickened cell walls were found. The results showed that the rapid decrease in temperature produced slender tracheids with obvious thickening of cell walls that resembled latewood cells. Conclusions The results suggest that a localized decrease in temperature of stems induces changes in the diameter and cell wall thickness of differentiating tracheids, indicating that cambium and its derivatives can respond directly to changes in temperature.     

Changes in diffusion resistance of apple leaves undergoing rapid fluctuations in leaf temperature

Transpiration was measured in apple leaves (Malus sylvestris Mill.) which were enclosed in a leaf chamber and subjected to rapid changes in leaf temperature. Fluctuations in leaf temperature produced parallel fluctuations in transpiration. The change in transpiration rate with change in temperature was found to be less than the theoretical value calculated from the change in water vapour density gradient from leaf to air. The results suggest the presence of a small and rapidly varying resistance to water vapour loss from the leaf. The magnitude of this additional resistance increased to a maximum value of approximately 1.5 s cm^-1 as the magnitude of the temperature change increased to a maximum of approximately 12°C.     

A high performance liquid chromatography assay for the rapid analysis of the subunit content of concanavalin A

A high performance liquid chromatography system is described which provides a rapid and convenient assay for the relative amounts of intact (26 000 dalton) and fragmented (14 000 and 12 000 dalton) subunits present in preparations of concanavalin A. Analyses were performed on an HPLC size exlusions column using either 8 M urea or 6 M guanidine hydrochloride as denaturing eluents. The efficiency and resolving power of this technique were confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. This HPLC assay facilitated the monitoring of the purification of concanavalin A to prepare a homogeneous preparation necessary for its biological evaluation.     

Breaks in arrhenius plots of reactions involving membrane-bound and solubilized sialyltransferases,due to temperature dependence of kinetic parameters

Temperature dependence of asialomucin-sialyltransferase ($\text{CMP}\text{-N-}\text{acetylneuraminate:}\text{D}\text{-galactosyl-glyco-protein}$) N-acetylneuraminyltransferase, EC 2.4.99.1) activity is investigated. Discontinuities in Arrhenius plots are observed, whether the enzyme is membrane-associated or solubilized. These discontinuities cannot be firmly correlated with the phase-transition temperatures of either endogenous or exogenous phospholipids. Arrhenius plots of the kinetic parameters also exhibit sharp discontinuities, so that it is concluded that a significant change in K_m and V_max values occurs with varying temperature. Our results suggest that the biphasic behavior of Arrhenius plots may be attributed to the temperature dependence of the kinetic parameters for both membrane-associated and solubilized sialyltransferase activities.