A simple method for determining total glucose utilisation by isolated adipocytes using (5-3H)-glucose

Glucose utilisation by adipocytes incubated with and without insulin and at two concentrations of extracellular glucose has been estimated by three different procedures. Glucose disappearance from the medium was calculated by using glucose oxidase to determine the glucose concentration remaining after incubation and comparing this with the glucose concentration in standard solutions made up by appropriate dilution of the original medium. [U-¹⁴C]-glucose utilisation was calculated by summing the ¹⁴C found in CO₂, triglycerides, and anions. [³H]-H₂O formation from [5-³H]-glucose was the third measure of glucose utilisation. All three methods gave similar answers, but the [5-³H]-glucose is simpler to use than [U-¹⁴C]-glucose and gives substantially more reproducible results than glucose oxidase.     

Effect of short schemes on body composition measurements using Air-Displacement Plethysmography

Background

Air-displacement plethysmography (ADP) is becoming a popular method to assess body composition. Several studies have shown certain types of clothing can affect measurements of body density, however no study has specifically investigated the effect of cotton gym shorts and spandex bicycle shorts on body density.

Methods

Thirty-seven males (23.0 ± 3.2 yr., 177.3 ± 5.4 cm., 74.8 ± 7.5 kg.) and thirty-eight females (23.7 ± 5.3 yr., 163.6 ± 8.4 cm., 57.1 ± 7.0 kg.) had their body density measured by ADP in three clothing schemes: 1) a tight fitting Speedo^® swim suit (criterion measure), 2) cotton gym shorts, and 3) spandex bicycle shorts. The clothing was provided by the University of Oklahoma Body Composition Laboratory and the testing schemes were performed in random order.

Results

The regression of body density by the criterion measure against body density while wearing cotton gym shorts for the entire group (y = 0.001 + 0.991x, SEE = 0.003 g/cm³) and the females (y = 0.059 + 0.934x, SEE = 0.003 g/cm³) did not significantly deviate from the line of identity. However in males the regression significantly deviated from the line of identity (y = 0.052 + 0.944x, SEE = 0.002 g/cm³). Body density by the criterion measure and body density while wearing spandex bicycle shorts did not significantly differ from the line of identity for the entire group (y = -0.018 + 1.013x SEE = 0.003 g/cm³), in males (y = -0.002 + 1.001x, SEE = 0.003 g/cm³), or females (y = 0.073 + 0.925x, SEE = 0.003 g/cm³). Residual plot analysis revealed no group or gender bias in either the cotton gym shorts or in the spandex bicycle shorts.

Conclusion

It would appear bicycle spandex shorts are an acceptable alternative to a Speedo^® like swim suit, however we advise that subjects adhere to the strict clothing protocol that is recommended by the manufacturer.

     

Carbonic anhydrase: a new method of detection on polyacrylamide gels using conductivity measurements

Carbonic anhydrase could be located on polyacrylamide gels by conductivity measurements using CO₂ as a substrate. The rapid increase of ions at the site of enzymatic localization caused an enhancement of conductivity which could be immediately recorded by the described technique.     

A new method for temperature compensation of electrical conductivity using temperature-fold dependency of fresh water

A new method for temperature compensation of electrical conductivity (EC) is proposed. A relationship between temperature and EC was investigated in detail by a simple experiment using natural and artificial fresh water. Results showed that the temperature dependency of EC varied from 0.03 to 0.02, with temperature increasing from 3°C to 35°C. This means that a traditional method for compensating for EC using a constant coefficient is invalid, so that a temperature-fold dependency must be taken into consideration to standardize EC to a common temperature.     

Evaluation of a method for determining the length of sperm whales (Physeter catodon) from their vocalizations

The click vocalizations of sperm whales often contain several regularly-spaced, discrete pulses of sound. Norris & Harvey (1972) hypothesized that these were caused as a single pulse of sound produced at the front of the whale's head bounced between reflective air sacs at either end of the spermaceti organ. Thus the interval between pulses will be twice the travel time for sound along the length of the spermaceti organ. It should therefore be possible to determine spermaceti organ length and thence total body length by measuring the interval between these pulses. Several workers have used an equation relating inter-pulse interval (IPI) to body length to estimate sperm whale body lengths acoustically.
In this paper, aspects of this technique are examined in some detail. In particular, variability in IPIs and trends in IPI with time and depth are investigated. Most importantly, for the first time IPIs in the vocalizations of whales of known lengths have been measured.
Variability in IPIs in the clicks of a single whale is acceptably low though there is a tendency for low and high values to occur in runs. There is no clear trend for IPI to alter significantly with the whale's depth or with the time since leaving the surface.
IPIs are positively correlated with body length though not as predicted by the equations used by previous workers. Some likely errors in these equations are discussed. A new empirically derived relationship between IPI and body length has been calculated, though more data are desirable to obtain a more accurate and reliable equation.     

Continuous measurements of electrical conductivity in growing media using a modified suction probe: Initial calibration and potential usefulness

A electrical conductivity (EC) cell introduced into a porous ceramic cup was developed to continuously sample the solution and measure EC from different growing media. Application of pressure head creates a continuous flow of solution from the growing media tested, into the ceramic cup, and through the EC cell. Continuous recording of the EC was achieved by connecting the EC meter to a data logger. Using two different pressure heads (–5 and –15 kPa) allowed us to observe differences in the EC of the solution extracted that resulted from the different moisture retention of each growing media. After a maximum period of 24 h extracting the solution from different growing media, EC values obtained with the probe were compared with those obtained using paste extracts, saturated with either deionized water or nutrient solution. Results show EC values obtained using the probe with a –15 kPa pressure head were closer to values of EC measured in saturated extracts made with nutrient solution. Using a –5 kPa pressure head, EC values with probe were lower than those obtained by extracts done with nutrient solution, but higher than EC values from saturated extracts made with deionized water. Simultaneous measurements of matric tension and EC show the effect of pressure heads applied in the probe on the water content of growing media. This technique is not destructive (the sampling of growing media is not necessary) and it is possible to obtain EC measurements of solutions continuously. This method of measuring water and salt content of the root environment has potential applications in the greenhouse production of pot plants. Measurements obtained with this method may lead to new information on nutrient uptake by plants and the development of new strategies of managing fertility and irrigation of horticultural crops.     

Determination of total body water by deuterium isotope dilution measurements using chemical ionization mass spectrometry

Two chemical ionization mass spectrometric methods were developed for direct determination of deuterium in water in the range of 0.0-0.6% 2H2O. One of them utilizes the batch inlet system, methane as the reagent gas, and the peak matching device of a magnetic sector mass spectrometer. The second one utilizes the directly-coupled gas chromatograph of a quadrupole mass spectrometer and computer control for ion selection and data processing. In this method the water itself serves as the reagent gas. The deuterium concentration is calculated from the ratio of ion intensities at m/z 20 (2HH2O+) and m/z 19 (H3O+). We have used these methods to determine total body water in 350 human subjects, which entailed making 900 measurements over a period of four years. Comparisons were made in 200 subjects of our results with those obtained by the creatinine method. No significant differences were found.     

Evaluation of a method for determining the free carboxyl group distribution in pectins

The distribution of free carboxyl groups in pectins has been investigated by a method that involves blocking the free carboxyl groups by glycolation, and hydrolysis of the methylesterified regions with a mixture of pectic enzymes. The hydrolysis products are separated from the glycolated regions on an ion exchange column and after deglycolation the oligomer size distribution is obtained by Sephacryl S-200 chromatography.The method was applied to five pectins with degrees of esterification in the range 5–70%. For two of the samples (an enzyme and an alkali de-esterified low methoxyl pectin) the degree of hydrolysis was significantly lower than would be predicted from the initial degree of esterification and thus for these materials the values obtained for the carboxyl group block sizes were considered to be a maximum rather than an accurate estimate.All the samples investigated had a significant proportion of free carboxyl regions with a degree of polymerisation greater than 10. With the possible exception of the pectate (degree of esterification 5%) none of the samples had a random distribution of carboxyl groups. This was considered to be a reflection of the distribution in the native pectin rather than indicating that chemical de-esterification was non-random. The large free carboxyl group block sizes was consistent with the egg-box model for low methoxyl pectin gelation. Larger blocks were found in the enzyme de-esterified pectin compared with the alkali and acid de-esterified material.     

A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer

A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES) cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture.     

Changes in egg and body temperature indicate triggering of egg desertion at a body mass threshold in fasting incubating blue petrels (Halobaena caerulea)

The triggering of transitory egg desertion in fasting and incubating blue petrels (Halobaena caerulea, nocturnal burrowing seabirds living in the subantarctic region) was investigated by continuously monitoring both body temperature (T _sto) and egg temperature (T _egg) with a telemetry system, and by measuring body mass (BM) loss. The birds were kept captive in their burrow and incubated day and night without any interruption; there was no day-night cycle in T _sto and T _egg, which averaged 39.9 °C and 32.0 °C, respectively. There was no evidence of hypothermia as a way to save energy in this fasting situation. Egg desertion occurred at night and was an abrupt and definitive phenomenon reflected by a simultaneous fall in T _egg and a peak in T _sto. After egg desertion, a distinct day-night cycle of body temperature was observed, T _sto being 0.6 °C higher during night-time (P < 0.05), probably reflecting increased nocturnal activity. BM at egg desertion averaged 166.7 ± 3.8 g in telemetered birds and 164.4 ± 1.6 g in␣a group of free-living birds. Throughout fasting, the␣specific daily BM loss remained at 46 ± 1 g · kg⁻¹ · day⁻¹, but increased sharply below a critical BM of 160.0 ± 2.5 g. Thus, fasting incubating blue petrels spontaneously desert their egg when reaching a BM threshold. This BM is very close to a critical value in fasting birds and mammals that corresponds to a critical depletion of fat stores and to a shift from lipid to protein utilization. This strongly suggests that such a metabolic shift triggers behavioural changes leading to egg desertion and refeeding, which is of great relevance to the understanding of the long-term control of food intake and BM. Accepted: 16 July 1998     

A rapid method for determining the fatty acid composition of lipid A

Lipid A is the most conservative part of LPS. Its fatty acids composition can serve as an important taxonomic marker of bacteria. The isolation of LPS and studying their chemical composition are difficult and protracted procedure. We propose the rapid method of determining the prevailed fatty acids of lipid A without isolation of LPS from the cell. The essence of the method is in the release of cell from the lipids which are not components of LPS. These lipids, in contrast to the lipid A, are more easily extractable from the cell structures. The fatty acids, which prevailed in the lipid-free cells, are the structural components of lipid A.     

A simplified method for determining 1-aminocyclopropane-1-carboxylic acid (ACC) in plant tissues using a mass selective detector

A sensitive and specific method is described for the routine assay of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) in 100–200 mg fresh weight samples of green or etiolated tissue. The method involves high performance liquid chromatography (HPLC) and gas chromatography linked to mass spectrometry (GCMS) and uses ¹⁴C-labelled ACC as an internal standard, N-benzoyl n-propyl ACC as an easily prepared derivative for HPLC and GCMS, and N-benzoyl isobutyl ACC as an internal standard for GCMS. The procedure is faster and safer than an existing GCMS method and more specific and reliable than indirect assays widely in use. The method has been used to measure ACC in maize roots, young leaves of cucumber, and aerobic or anaerobic seedlings of rice.