Improved fluorometric assay of histamine: condensation with O-phthalaldehyde at -20 degrees C

Histamine reacts with OPT at an alkaline pH giving rise to fluorescent conjugation products. Optimum fluorophore formation was observed at pH 12.5 after 10 hr under nitrogen at ?20°C, i.e., in the frozen state. After acidification with sulfuric acid to pH 2.5 the resulting fluorescence, read at room temperature, was stable for hours. The procedure now measures as little as 1 ng histamine/ml and is much more specific than the conventional fluorometric assay. Spermidine did not interfere with the assay of histamine, and histidine only if present in great excess over histamine. It could be shown that with deproteinized extracts of rat gastric mucosa, histamine could be estimated without further purification, which means saving a lot of time and labor.     

Finger temperatures during military field training at 0 to −29 °C

1. Skin and rectal temperatures were recorded continuously in 70 measurements during typical tasks of infantry and artillery training at 0 to −29 °C. The duration of the measurements varied from 55 min to 9.5 h.

2. The distribution of finger skin temperatures was quite similar at ambient temperature ranges 0 to −10 °C and −10 to −20 °C, while at −20 to −30 °C the finger temperatures were clearly lower.

3. At different ambient temperature ranges, 20–69% of finger temperatures were low enough to cause cold thermal sensations.

4. Sensation of cold was experienced at a finger temperature of 11.6±3.7 °C (mean±SD).     

Proline reductase: a sensitive fluorometric assay with O-phthalaldehyde.

A rapid and sensitive fluorometric assay for measuring the activity of proline reductase is described. The product of the enzymic reaction, δ-aminovalerate, is converted to a highly fluorescent derivative by reaction with o-phthalaldehyde. The water-soluble fluorescent product exhibits an excitation maximum at 340 nm and an emission maximum at 455 nm. The fluorophor reacts specifically with δ-aminovalerate without any interference from proline.     

Mechanical properties of the human red blood cell membrane at −15 °C

The most common method for measuring the mechanical behavior of the human red blood cell (RBC) membrane is micropipette aspiration, because it can be used to apply both a low uniaxial stress at a small part of the membrane or high two-axial stresses to the whole membrane [E.A. Evans, R.E. Waugh, Mechano-chemical study of red cell membrane structure in situ, in: Kroc Foundation Series, vol. 13, Erythrocyte Mechanics and Blood Flow, Alan R. Liss. Inc., New York, 1980, pp. 31–56 (Chapter 3); H.J. Meiselman, Measures of blood rheology and erythrocyte mechanics, in: Kroc Foundation Series, vol. 13, Erythrocyte Mechanics and Blood Flow, Alan R. Liss. Inc., New York, 1980, pp. 75–117 (Chapter 5)]. The elastic shear moduli and area changes of the human RBC published to date were calculated by means of this technique. However, a main drawback of the method is its impracticability at subzero temperatures. Experiments at below 0 °C are of interest because it is at these temperatures that RBC lysis occurs during freezing and thawing after cryopreservation, via a mechanism that may be mechanical.A method for circumventing this limitation is deforming the cell membranes by applying an electric ac field to a supercooled suspension. In a previous study, we applied this technique to human RBCs down to −15 °C [M. Krueger, F. Thom, Deformability and stability of erythrocytes in high-frequency electric fields down to subzero temperatures, Biophys. J. 73 (1997) 2653–2666]. In this technique, the electrical dimensions must be translated into those of mechanics. We provided a formula for these calculations, which demonstrated excellent concordance with known mechanical measurements at room temperature [F. Thom, H. Gollek, Calculation of mechanical properties of human red cells based on electrically induced deformation experiments, J. Electrostat. 64 (2006) 53–61]. Using this formula, we have now calculated the shear moduli and stress–strain diagram for our deformation experiments at −15 °C and present the results below.     

Effects of metabolic rate on thermal responses at different air velocities in −10°C

The effects of exercise intensity on thermoregulatory responses in cold (−10°C) in a 0.2 (still air, NoWi), 1.0 (Wi1), and 5.0 (Wi5) m s⁻¹ wind were studied. Eight young and healthy men, preconditioned in thermoneutral (+20°C) environment for 60 min, walked for 60 min on the treadmill at 2.8 km/h with different combinations of wind and exercise intensity. Exercise level was adjusted by changing the inclination of the treadmill between 0° (lower exercise intensity, metabolic rate 124 W m⁻², LE) and 6° (higher exercise intensity, metabolic rate 195 W m⁻², HE). Due to exercise increased heat production and circulatory adjustments, the rectal temperature (T_re), mean skin temperature (T_sk) and mean body temperature (T_b) were significantly higher at the end of HE in comparison to LE in NoWi and Wi1, and T_re and T_b also in Wi5. T_sk and T_b were significantly decreased by 5.0 m s⁻¹ wind in comparison to NoWi and Wi1. The higher exercise intensity was intense enough to diminish peripheral vasoconstriction and consequently the finger skin temperature was significantly higher at the end of HE in comparison to LE in NoWi and Wi1. Mean heat flux from the skin was unaffected by the exercise intensity. At LE oxygen consumption (V ₂) was significantly higher in Wi5 than NoWi and Wi1. Heart rate was unaffected by the wind speed. The results suggest that, with studied exercise intensities, produced without changes in walking speed, the metabolic rate is not so important that it should be taken into consideration in the calculation of wind chill index.     

Thermal responses during and after whole-body cryotherapy (−110°C)

The effect of whole-body cryotherapy (WBC) on rectal and skin temperatures was measured in healthy subjects before, during and after WBC exposure. WBC did not cause any significant change in rectal temperature. The lowest local skin temperatures were recorded in the forearm, 5.2 (2.8)°C, and in the calf, 5.3 (3.0)°C. WBC involves no risk for frostbites. After WBC, all skin temperatures recovered rapidly, indicating that the analgetic effects of WBC only occur during a limited period after the exposure.     

Water lecithin binding in lecithin-water lamellar phases at 20°C

¹H and ³¹P continuous wave and spin-echo NMR measurements have been made on lecithin-water mixtures as a function of water content at 20 °C. An analysis of the data demonstrates the existence of two water environments, lecithin bound and free. Conclusions are presented concerning the stoichiometry and kinetics of the binding. The results indicate that six molecules of water are bound to one lecithin molecule and the lifetime of the bound water is 6·10⁻⁵±3·10⁻⁵ s.     

Algae Permeability to Me2SO from −3 to 23°C

Biphasic transport of water and dimethyl sulfoxide (Me₂SO), a common cryoprotective agent (CPA), in algal cells was induced and measured on a cryoperfusion stage. A two-step experimental protocol provided data for the volumetric response of Chlorococcum (C.) texanum to impermeable and permeable solutes. First, the cells were exposed to a 500-mOsm sucrose solution, causing immediate shrinkage of the cell to a minimum equilibrium volume. Then an isoosmotic 200-mOsm/300-mOsm CPA/sucrose solution was introduced to the cells, resulting in increased cell volume to a new equilibrium state. Experiments were conducted at temperatures between −3 and 23°C. Cell volumes were measured off-line by computer analysis of video images. A network thermodynamic model was fit to the transient volume data to determine permeabilities of C. texanum to water and Me₂SO over the full temperature range, and results were calculated with two numeric methods. Biphasic transport was found to be slower at colder temperatures, with water entering the cell faster than Me₂SO. Experimental results were also compared with data from similar experiments using methanol (MeOH) as the CPA. MeOH influx was calculated to be a magnitude larger than that of water. Additionally, MeOH permeability was at least three orders of magnitude greater than Me₂SO permeability, and the difference in these solute permeabilities increased as temperature decreased.     

The effect of cryopreservation at −196°C on the viability of fowl and Turkey spermatozoa assessed in vitro

The effect of storing fowl and turkey spermatozoa at −196°C in a glycerol-based diluent on post-thawing levels of motility, ATP content, morphology and leakage of lactate dehydrogenase was investigated. Compared to those of freshly-diluted spermatozoa, these parameters were reduced to 24, 26, 55 and 61% respectively, for fowl spermatozoa and to 22, 26, 34 and 60%, respectively, for turkey spermatozoa, immediately after thawing and the removal of glycerol. On subsequent incubation for 3 h at 40°C, the ATP content and morphological integrity of frozen/thawed spermatozoa of both species fell to less than 5% of the levels of similarly-incubated non-frozen spermatozoa. Only after prolonged incubation of fowl and turkey spermatozoa could these parameters be shown to reflect, and, therefore, be of use as monitors of the reduction of fertilizing ability caused by freezing.     

Comparisons of blood viscosity between amphibians and mammals at 3°C and 38°C

(1) The range of temperature exposure of endotherms is narrow compared to ectotherms that can experience daily and seasonal temperature fluxes. (2) Comparison of the blood viscosity of amphibians (bullfrog, Woodhouse's toad, and marine toad) and mammals (horse, dog, and rat) at 3°C and 38°C was undertaken to determine if the effect of temperature on blood viscosity was diminished in amphibians relative to mammals. (3) Mammals did not consistently show greater changes in blood viscosity, plasma viscosity, or relative viscosity with decreasing temperatures relative to the amphibians in this study. (4) These data do not support our hypothesis that blood viscosity of amphibians is less affected by temperature than mammalian blood.     

Antigen rearrangements in Colpoda maupasi cells after freezing at −196 °C, and after shortwave ultraviolet irradiation

Changes in the antigenic content of resting cysts of ciliates after freezing at −196 °C have been studied. The results obtained from experiments with native immunosera using the immobilizing and agglutinating activity allow the conclusion that freezing causes significant changes in the cyst surface antigens. Ultraviolet irradiation did not cause any notable shifts in the surface antigen composition.By means of immunodiffusion methods, changes in the soluble antigen set of cysts after freezing have been demonstrated. Freezing induces a specific simplification in the antigen spectrum and a new antigen appearance characteristic of cysts subjected to freezing.     

Improved Control of Otiorhynchus sulcatus at 9°C by Cold-stored Heterorhabditis megidis UK211

The effect of storage temperature (9 and 20°C) on North West European Heterorhabditis megidis isolate UK211 for control of Otiorhynchus sulcatus larvae at 9°C is assessed. O. sulcatus mortality increased from -5.3% (corrected mortality) using freshly produced nematodes, to 27.1% using nematodes that had been cold-stored for 12 weeks. The number of nematodes invading the insect larvae increased almost 27-fold. Nematode storage at 9°C for 11 to 12 weeks weeks resulted in significantly higher O. sulcatus mortality (41%) than storage at 20°C for 2 to 3 weeks (12%). Thus, cold storage does enhance nematode infectivity for O. sulcatus larvae.