Determination of the polymorphic composition of smooth pea starch

A method for estimating the proportions of ‘A’ and ‘B’ polymorphs comprising a sample of ‘C’ type starch is proposed which uses established experimental techniques with commercially available spreadsheet and X-ray analysis software. Waxy maize, potato and smooth pea starches were used to provide X-ray diffraction patterns characteristic of the ‘A’, ‘B’ and ‘C’ starch polymorphs. Samples of amorphous starches were also prepared. The method initially involved subtraction of the amorphous phase and instrumental background from the X-ray diffraction patterns of each starch sample using the spreadsheet program, Lotus 1-2-3. The remainder of the pattern, representing the crystalline portion of the starch sample, was then analysed by profile fitting to elucidate the positions and areas of individual diffraction peaks. The ratio of the total peak area to the areas under peaks characteristic of ‘A’ and ‘B’ type starches, respectively, were used to calculate the relative proportions of these polymorphs in smooth pea starch. These proportions were found to be 56±3% ‘A’ polymorph to 44±3% ‘B’ polymorph. A ‘C’ type pattern was constructed by using Lotus 1-2-3 to combine diffraction patterns from the crystalline portions of ‘A’ and ‘B’ type starches in the proportions given above. Polymorph patterns were obtained by manipulation of the diffraction patterns from the crystalline portions of starches using Lotus 1-2-3. An ‘A’ type pattern was obtained by subtraction of a ‘B’ type pattern from that of a ‘C’ type. Similarly, a ‘B’ type pattern was obtained by subtraction of an ‘A’ type pattern from that of a ‘C’ type.     

Intracellular signalling proteins as ‘smart’ agents in parallel distributed processes

In eucaryotic organisms, responses to external signals are mediated by a repertoire of intracellular signalling pathways that ultimately bring about the activation/inactivation of protein kinases and/or protein phosphatases. Until relatively recently, little thought had been given to the intracellular distribution of the components of these signalling pathways. However, experimental evidence from a diverse range of organisms indicates that rather than being freely distributed, many of the protein components of signalling cascades show a significant degree of spatial organisation. Here, we briefly review the roles of ‘anchor’, ‘scaffold’ and ‘adaptor’ proteins in the organisation and functioning of intracellular signalling pathways. We then consider some of the parallel distributed processing capacities of these adaptive systems. We focus on signalling proteins-both as individual ‘devices’ (agents) and as ‘networks’ (ecologies) of parallel processes. Signalling proteins are described as ‘smart thermodynamic machines’ which satisfy ‘gluing’ (functorial) roles in the information economy of the cell. This combines two information-processing views of signalling proteins. Individually, they show ‘cognitive’ capacities and collectively they integrate (cohere) cellular processes. We exploit these views by drawing comparisons between signalling proteins and verbs. This text/dialogical metaphor also helps refine our view of signalling proteins as context-sensitive information processing agents.     

Biosynthesis of Hb in individual fetal liver bursts : γ-Chain production peaks earlier than β-chain in the erythropoietic pathway

The relative synthesis of α-, β-, ^Gγ- and ^Aγ-globin chains has been evaluated in single fetal liver bursts, which were grown in methylcellulose cultures, individually labelled with [³H]leucine and then analysed via iso-electric focusing. Well-hemoglobinized bursts demonstrate a homogeneous globin synthetic pattern, characterized by prevalent HbF (+some HbA) synthesis: thus, they apparently originate from a homogeneously programmed population of erythroid burst-forming unit (BFU-E). On day 8–9 of culture, the synthetic pattern in ‘mature’ (i.e., well-hemoglobinized) bursts has been compared with that in simultaneously-grown, ‘immature’ (i.e., poorly-hemoglobinized) colonies. These patterns have been further compared with that in ‘matured’ bursts (identified in situ as immature on day 8–9 and labelled 2–4 days later when matured). The ‘immature’ colonies showed very low levels of relative β-globin synthesis, while the ‘mature’ ones demonstrated a more elevated production of β-chain. Significantly, the ‘matured’ bursts showed a globin chain synthetic pattern similar to that of previously labelled ‘matured’ colonies. It is postulated therefore that in fetal liver (and also in adult marrow) the synthesis of γ-chain is linked to an early differentiation stage of erythroblasts, while β-globin synthesis is largely activated at a more advanced maturation stage.     

Suppression of the transformed phenotype with retention of the viral ‘src’ gene in cell hybrids between rous sarcoma virus-transformed rat cells and untransformed mouse cells

Hybrid cell lines between untransformed mouse 3T3TK-cells and normal rat kidney (NRK) cells transformed by the B77 strain of Rous Sarcoma Virus (RSV) express a non-transformed phenotype, as determined by anchorage-dependent growth and organization of microfilament bundles. Virus rescue experiments and genetic experiments using an RSV mutant temperature-sensitive for maintenance of the transformed phenotype demonstrate that RSV is retained in the non-transformed hybrids. The action of the viral transformation gene ‘src’ therefore appears to be ‘suppressed’ in these hybrids. The suppressed hybrids generate variants in which the expression of the transformed phenotype and the ‘src’ gene is regained. This system should prove to be of value in identifying cellular genes involved in the expression of virally induced transformation.     

Rapid multichannel microfluorometry for metabolic and spectral studies in single living cells

A multichannel microspectrofluorometer has been developed for operation on two modes, a ‘morphological’ mode for the assay of intracellular fluorochromes (e.g. NAD(P)H) in correlation with topography, and a ‘spectral’ mode for wavelength analysis of natural cell fluorescence. This instrument is based on an electron bombardment silicon camera tube (EBS) operated in conjunction with a multiscaling computer. The total NAD(P)H emission from 2 × 30 μm cell strips can be analysed in real time (32.8 msec frame scan) with a signal-to-noise ratio over 100:1. The metabolic changes in cytoplasmic regions are compared with those in regions comprising cytoplasm + nucleus, where the major contribution may be nuclear (cf earlier studies). The observation of a ‘multilocalized’ and asynchronous metabolic response is facilitated with substrates such as glucose-1-phosphate, associated with a longer lag period before the initiation of fluorescence changes. The latter largely occur in the 440–480 nm region. Fluorescence spectra recorded from intracellular regions are nearly super-posable to the spectrum obtained from NAD(P)H crystals.     

The regulatory function of the myosin light chains

In molluscan muscles, calcium regulation is mediated by ‘regulatory’ light chains associated with the myosin heads. This type of ‘regulatory’ light chain appears to be present in all myosins, regardless of whether the myosin contains light chain linked calcium regulation. Although they appear to be ‘structurally’ related, differences in their calcium binding abilities imply that these regulatory light chains may play quite distinct functions in their respective myosins.     

Double infection experiments with echinostomatids (Trematoda) in Lymnaea stagnalis by implantation of rediae and exposure to miracidia

1972. Double infection experiments with echinostomatids (Trematoda) in Lymnaea stagnalis by implantation of rediae and exposure to miracidia. International Journal for Parasitology, 2: 409–423. Echinostomatid species parasitizing Lymnaea stagnalis as first intermediate hosts in a South German Lake have been found present in natural double infections, but at frequencies lower than expected. Simultaneous double infection and superinfection experiments in Lymnaea stagnalis with Isthmiophora melis, Echinoparyphium aconiatum and Echinostoma revolutum were performed by redial implantation and by exposure to miracidia. All three combinations possible of these echinostomatids proved to be unstable, one species being eliminated by another ‘stronger’ one after an invariable suppression order. The degree of vigour of Isthmiophora melis in this suppression order is greater if mother rediae (macropharyngeate) are present, i.e. after miracidial invasion instead of daughter redial implantation. Snails parasitized by rediae of a ‘weak’ type could be superinfected by implantation of rediae of a ‘strong’ type, but not if the first (‘weak’) infection had reached the stage of shedding cercariae. Superinfection by implantation of Echinoparyphium aconiatum rediae (‘strong’ type) was not successful when the first infection consisted of sporocysts of plagiorchiids, or of Apatemon sp. (Strigeidae) that had reached the stage of shedding cercariae.     

The cooperativity phenomena in a pigment-protein complex of light-harvesting antenna revealed by picosecond absorbance difference spectroscopy

A model of the cooperative changes in optical properties of light-harvesting bacteriochlorophyll molecules of complex B890 in response to the absorption of light quanta is proposed. According to the model, each antenna chromophore may persist in either of two optically non-excited states, R and T. The occurrence of at least one excitation per complex causes all optically non-excited chromophores of the complex to be converted from state R to state T. The theory is shown to be in good agreement with experimental ‘light curves’ (ΔAvs intensity of picosecond excitation pulse) for the ‘minor’ and ‘major’ signals of light-harvesting bacteriochlorophylls of complex B890 from Chromatium minutissimum.     

Different architectures of the collagen fibril: morphological aspects and functional implications

Several tissues known to contain collagen fibrils with a ‘helical’ arrangement were studied by t.e.m. and freeze-fracture. In all the tissues examined, the diameter of the collagen fibrils appeared to be tissue-specific and fairly constant within the same tissue. No statistical differences, on the contrary, were detectable in the coiling angle which appeared similar in all the tissues and independent of both diameter and age of the fibril. Rat tail tendon was also examined under the same technical conditions and showed collagen fibrils of large and very heterogeneous diameter and with a consistent ‘straight’ arrangement. These data seem to suggest that the ‘helical’ and ‘straight’ arrangements may actually identify different types of collagen fibrils. The authors discuss the possible functional significance of these arrangements and present two hypotheses on the three-dimensional structure of the ‘helical’ fibril.     

Spinach-thylakoid phosphorylation: Studies on the kinetics of changes in photosystem antenna size, spill-over and phosphorylation of light-harvesting chlorophyll a/b protein

The kinetics of LHCP phosphorylation and associated changes in photosystem cross-section and energy ‘spill-over’ from PS II to PS I have been examined in isolated spinach chloroplasts. During an initial phosphorylation period of 3–6 min, in the presence of saturating concentrations of Mg²⁺, the increase in PS I and decrease in PS II cross-section are largely completed, as judged by both measurements of the steady-state redox state of Q and fluorescence yield changes. This corresponds to a period of rapid ³²P incorporation into the low-molecular weight LHCP polypeptide. Subsequent to this initial 3–6-min period there is substantial further phosphorylation of both LHCP polypeptides, which is not accompanied by significant changes in photosystem cross-section, even after the chloroplasts had been unstacked with extensive mixing of PS I and PS II by Mg-removal. It is suggested that there exists a specific ‘mobile’ population of LHCP molecules which is rapidly phosphorylated and which may be enriched in the low-molecular-weight polypeptide. In addition, measurements of the kinetics of the ‘spill-over’ changes upon either Mg²⁺ addition or removal indicate that the continued phosphorylation of LHCP is able to increase the ‘spill-over’ process under favourable ionic conditions.     

Formation of triton WR 1339-filled rat liver lysosomes : II. Involvement of autophagy and of pre-existing lysosomes

The participation of endocytosis in the formation of Triton-filled lysosomes was followed after injecting Triton WR 1339 simultaneously with colloidal gold or horseradish peroxidase. According to cytomorphometric measurements Triton WR 1339 also significantly enhances autophagy. The analysis of the influence of Triton WR 1339 subfractions shows that autophagy is only augmented by the polymer component (‘macrotriton’) but not by the low molecular component (‘microtriton’) alone. When the latter was injected combined with either macrotriton or colloidal gold particles, autophagy increased to a level observed with Triton WR 1339 and beyond the levels determined for either component alone. Thus, autophagy appears to be stimulated by endocytosis. Autolysosomes are transformed within a few hours to peribiliary ‘dense bodies’. These do not display any significant turnover rate within a period of several days; therefore, dense bodies could be prelabelled with colloidal gold in order to show that they are the target organelles ingesting (macro-)Triton. According to difference spectra autophagy leads to a considerable concentration of microsomal and mitochondria! cytochromes in (macro-)Triton-filled lysosomes far beyond the level detected in ‘normal’ lysosomes. Because of the participation of both hetero- and autophagy in the formation of Triton WR 1339-filled lysosomes they have to be classified as telolysosomes.     

Transitions between cooperative and non-cooperative responding in the 'Pigeon's Dilemma'

Two pigeons played Iterated Prisoner's Dilemma (IPD) against a simulated opponent pre-programmed to play ‘Tit-For-Tat’ (TFT) and ‘Random’ (RND) strategies. Each pigeon received differential amounts of access to food following choices of either ‘cooperate’ or ‘defect’ on a trial. After 1000 trials against TFT and 500 trials against RND, results indicated that choice allocation was optimal when the birds played against RND but was sub-optimal when the birds played against TFT. In order to determine why the pigeons responded suboptimally against TFT, a trial-by-trial analysis of the data was conducted. The analysis revealed that once a pigeon had received the ‘Sucker's’ payoff (S), it was more likely to defect and receive the ‘Temptation’ payoff (T) than to cooperate and receive the ‘Reward’ (R) payoff. Local reinforcement contingencies appear to determine suboptimal responding against TFT in the iterated Pigeon's Dilemma.     

Protein turnover and cell-cycle initiation in yeast

The effects of short pulses of cycloheximide on the traversal of the G1 phase of the cell cycle of the yeast Saccharomyces cerevisiae were examined. Cells were released from a block at the regulatory stage of G1, termed ‘start’, and pulsed with cycloheximide. Delays in budding which were considerably longer than the length of the pulse were observed. During the delay the cells remained blocked at ‘start’. No delay in budding was observed after cycloheximide pulses, when cells were released from a cdc 24 block which arrests the budding process but not ‘start’. Overall protein synthesis did not show an additional delay after the pulse. The extra lag following cycloheximide pulses appears to reflect a unique feature of ‘start’. It may be accounted for by a requirement at ‘start’ for a labile protein with a half-life time of about 6 min.     

Photoacoustic detection of photosynthetic activities in isolated broken chloroplasts

Methodology and demonstration how to utilize the photoacoustic technique in photosynthesis research are presented. Photoacoustic signals were obtained from suspensions of isolated broken chloroplasts. In the presence of strong, continuous (non-modulated) background light the signals were normally larger than without the background light. The effect of the background light was saturable and was absent when non-active (e.g. heat-treated) samples were used, showing that the normal smaller signal in the absence of background light is a genuine reflection of the loss of heat due to the competing photochemistry. The effect of the background light is to close the reaction-centers and hence to inhibit the photochemical process. The percent difference of the photoacoustic signal (± background light) is taken as a measure of the photochemical activity (‘photochemical loss’).

Initial results demonstrate the wavelength dependence of the ‘photochemical loss’. As expected there was a ‘red-drop’ decrease of the ‘photochemical loss’ for λ > 690 nm, when the cofactor methyl viologen was present. Surprisingly, however, there was a ‘red-rise’ increase for λ > 690 nm when no cofactor was present. These findings indicate that under the last conditions there is an unsuspected photoactivity of PS I which was not detected hitherto by the conventional techniques. The dependence on the background light intensity confirms this result. This photoactivity can be explained tentatively as a cyclic electron flow around PS I, present without any added cofactor.

Initial results on the modulation frequency dependence in the presence of electron acceptors are also demonstrated.     

Quantification of high-frequency sea-level fluctuations in shallow-water carbonates: an example from the Berriasian–Valanginian (French Jura)

When quantifying sedimentary processes on shallow carbonate platforms, it is important to know the high-frequency accommodation changes through time. Accommodation changes in cyclic successions are often analysed by simply converting cycle thickness to Fischer plots. This approach is not satisfactory, because it does not account for differential compaction, possible erosion, sea-level fall below the depositional surface, or subtidal cycles. An attempt is made here to reconstruct a realistic, high-frequency accommodation and sea-level curve based on a detailed facies and cyclostratigraphical analysis of Middle Berriasian to Lower Valanginian sections in the French Jura Mountains. The general depositional environment was a shallow-marine carbonate platform on a passive margin. Our approach includes the following steps: (1) facies interpretation; (2) cyclostratigraphical analysis and identification of Milankovitch parameters in a well-constrained chronostratigraphic framework; (3) differential decompaction according to facies; (4) estimation of depth ranges of erosion and vadose zone; (5) estimation of water-depth ranges at sequence boundaries and maximum flooding intervals; (6) estimation of mean subsidence rate; (7) classification of depositional sequences according to types of facies evolution: ‘catch-up’, ‘catch-down’, ‘give-up’, or ‘keep-up’; (8) classification of depositional sequences according to long-term sea-level evolution: ‘rising’, ‘stable’, ‘falling’; (9) calculation of ‘eustatic’ sea-level change for each depositional sequence using the parameters inferred from these scenarios, assuming that sea-level cycles were essentially symmetrical (which is probable in Early Cretaceous greenhouse conditions); (10) calculation of a sea-level curve for each studied section; (11) comparison of these curves among each other to filter out differential subsidence; (12) construction of a ‘composite eustatic’ sea-level curve for the entire studied platform; (13) spectral analysis of the calculated sea-level curves. Limitations of the method are those common to every stratigraphic analysis. However, the method has the potential to improve the original cyclostratigraphical interpretations and to better constrain the high-frequency sea-level changes that control carbonate production and sediment fluxes.     

A characteristic pharmacological action of ‘Yang-invigorating’ Chinese tonifying herbs: Enhancement of myocardial ATP-generation capacity

In order to investigate the pharmacological basis of ‘Yang-invigorating’ action, the effect of oral treatment with the methanolic extract of ‘Yang-invigorating’ herbs on ATP-generation capacity was examined, using heart homogenates prepared from herb-pretreated mice. Tonifying (i.e., health-promoting) herbs of other functional categories were also included for comparison. The results indicated that ‘Yang-invigorating’ Chinese tonifying herbs could invariably enhance myocardial ATP-generation capacity, with the extent of stimulation varying among the herbs. In contrast, ‘Yin-nourishing’ herbs either did not stimulate or even decreased myocardial ATP-generation capacity. While ‘Qi-invigorating’ herbs produced variable effects on myocardial ATP-generation capacity, most of the ‘blood-enriching’ herbs did not cause any significant changes. The results obtained from studies using myocardial mitochondrial fractions isolated from herb-pretreated mice suggest that ‘Yang-invigorating’ herbs might speed up ATP generation by increasing mitochondrial electron transport. The ensemble of results has provided evidence for the first time to support the pharmacological basis of ‘Yang invigoration’ in Chinese medicine.     

Comparative assessment of two algorithms for calibrating stereophotogrammetric systems

In this paper, a comparison is carried out between two photogrammetric algorithms aimed at camera calibration and three-dimensional target point reconstruction by ‘absolute’ distributions of control points; the first is Marzan and Karara's DLT, the second is CESNO, by the author, an algorithm quite close to Hatze's modified DLT (MDLT). The comparative assessment is especially aimed at testing the capability of the two methods to produce good results when calibration data are to be extrapolated beyond the space spanned by the control distribution. The assessment was carried out not by a real stereophotogrammetric system, but by the computer simulation of a two-camera set-up. Various combinations of internal camera parameters, such as the scaled principal distances, were tried out. As for the magnitude of the simulated non-linear lens distortion, three configurations were used which produced ‘low’, ‘medium’ and ‘strong’ distortion. The influence of decreasing the number of control (calibration) points on the accuracy performance of the two algorithms was also investigated. The results show the superiority of CESNO, especially with medium or strong lens distortion, or when the two camera principal distances sensibly differ.     

Contamination effects by a ‘conventional’ and a ‘biodegradable’ lubricant oil on infaunal recruitment to Antarctic sediments: A field experiment

To investigate the impacts of synthetic lubricants on Antarctic infaunal communities, a field experiment was setup near Australia's Casey Station, East Antarctica. Two types of synthetic lubricants were tested: an ‘Unused’ and ‘Used’ conventional synthetic lubricant, and an alternative marketed as ‘biodegradable’. Clean defaunated sediment was contaminated with the lubricants, decanted into trays, and deployed by divers onto the seabed in a randomised block design. Sediments were sampled 5 and 56 weeks after deployment. After 5 weeks, benthic assemblages that had recruited to the lubricant contaminated sediments were significantly different to those in ‘Control’ sediments, and differences were more pronounced after 56 weeks. Total number of individuals did not significantly differ between treatments after 5 weeks. However, after 56 weeks total individuals in the ‘Control’ sediments were significantly greater than in the contaminated sediments. Nototanais antarcticus (tanaid) and to a lesser extent Monoculodes sp. (gammarid), Tanaid sp. IV and Eudorella sp. (cumacean) had significantly higher abundances in the control sediments after 56 weeks compared to the contaminated sediments. Copepods numerically dominated the benthic assemblages at both sampling times; however, their abundance did not significantly differ across treatments. The community recruiting to the contaminated sediments remained different from that in the ‘Control’ sediments for the duration of the experiment (1 year). The ‘biodegradable’ lubricant was just as environmentally harmful to the Antarctic infauna as the ‘conventional’ lubricant currently used at Australia's Antarctic stations. Our results demonstrate that changes to recruitment are one of the potential environmental consequences of a lubricant spill to Antarctic benthic communities, and reinforce the importance of preventative oil spill management and effective clean-up procedures. Further monitoring of this field experiment will provide much needed information about the long-term impacts by synthetic lubricants in the Antarctic marine environment.