Pacemakers in aggregation fields of Dictyostelium discoideum: does a single cell suffice?

In this paper we address the following question: can a single cell of the cellular slime mold Dictyostelium discoideum serve as a pacemaker for the aggregation phase? Whether or not this is possible is determined by the relative importance of cyclic AMP production due to self-stimulation as compared to diffusion of cyclic AMP away from the cell and extracellular degradation. We determine the conditions under which a single cell on an infinite place can emit periodic signals of cyclic AMP using a model developed previously for signal relay and adaptation in Dictyostelium. Elsewhere it has been shown that this model provides an accurate representation of the stimulus-response behavior of Dictyostelium for a variety of experimental conditions.     

Interleukin-3-stimulated haemopoietic stem cell proliferation. Evidence for activation of protein kinase C and Na+/H+ exchange without inositol lipid hydrolysis.

Interleukin 3 (IL-3) is an important regulator of haemopoietic stem cell proliferation both in vivo and in vitro. Little is known about the possible mechanisms whereby this growth factor acts on stem cells to stimulate cell survival and proliferation. Here we have investigated the role of intracellular pH and the Na+/H+ antiport in stem cell proliferation using the multipotential IL-3-dependent stem cell line, FDCP-Mix 1. Evidence is presented that IL-3 can stimulate the activation of an amiloride-sensitive Na+/H+ exchange via protein kinase C activation. IL-3-mediated activation of the Na+/H+ exchange is not observed in FDCP-Mix 1 cells where protein kinase C levels have been down-modulated by treatment with phorbol esters. Also the protein kinase C inhibitor H7 can inhibit IL-3-mediated increases in intracellular pH. This activation of Na+/H+ exchange via protein kinase C has been shown to occur with no measurable effects of IL-3 on inositol lipid hydrolysis or on cytosolic Ca2+ levels. Evidence is also presented that this IL-3-stimulated alkalinization acts as a signal for cellular proliferation in stem cells.     

AUTUMN MIGRATION IN SOUTHWEST PORTUGAL.

During the latter half of September observations were made at various points on the coast of southwest Portugal. A great many migrants were seen, especially close to Cape St. Vincent, belonging to species that winter only south of the Sahara. Bird-catchers were in evidence on the south coast, but not on the west. Evidence was obtained that migrants are fewest with east winds
Most of the birds seen must have been travelling in a southwesterly direction for a long way and it is necessary for their survival that they should change course on reaching the shores of the Atlantic. Any that did not do so might find a temporary refuge on the Canary Islands, but the list of regular passage-migrants there is very different from that of the birds seen commonly near Cape St. Vincent     

Superoxide Dismutase as an Anaerobic Polypeptide : A Key Factor in Recovery from Oxygen Deprivation in Iris pseudacorus?

The perennating organ, the rhizome, was chosen for examination of response to anoxia in the species Iris pseudacorus L., Iris germanica L. var Quechei, and Glyceria maxima (Hartm.) Holmberg. These monocots are known to differ in their tolerance of anoxia. Intact rhizomes were subjected to periods of prolonged anoxia of up to 28 days and superoxide dismutase (SOD) activity was determined in a 48 hour postanoxic recovery phase. Tests were performed to ensure the accuracy of the measured enzyme activities. In the most anoxia tolerant species, I. pseudacorus, SOD activity rose continuously during the period of imposed anoxia, and levels were maintained in the postanoxic recovery phases: 28 days brought about a 13-fold increase to 1576 U SOD per milligram protein. Small increases were found in the less anoxia tolerant I. germanica during anoxic/postanoxic phases, while a drop in activity was recorded in the least anoxia tolerant G. maxima. However, initial levels in G. maxima were more than twice as high as in the other two species. Experiments applying cycloheximide to anoxic rhizome slices of I. pseudacorus inhibited the increase in SOD activity. This indicates that SOD is, paradoxically, induced under anoxia and we suggest that in this species SOD is one of the enzymes identified as anaerobic polypeptides. The significance of the induction of an `oxygen-protecting' enzyme during complete oxygen deprivation is discussed with regard to a possible critical role during recovery from anoxic stress.     

Bacteriophage applications: where are we now?

Bacteriophages are bacterial viruses and have been used for almost a century as antimicrobial agents. In the West, their use diminished when chemical antibiotics were introduced, but they remain a common therapeutic approach in parts of eastern Europe. Increasing antibiotic resistance in bacteria has driven the demand for novel therapies to control infections and led to the replacement of antibiotics in animal husbandry. Alongside this, increased pressure to improve food safety has created a need for faster detection of pathogenic bacteria. Hence, there has been a resurgence of interest in bacteriophage applications, and this has encouraged the emergence of a large number of biotech companies hoping to commercialize their use. Research in Europe and the United States has increased steadily, leading to the development of a range of applications for bacteriophage agents for the healthcare, veterinary and agricultural sectors. This article will attempt to answer the question of whether bacteriophages are now delivering on their potential.     

Passive multivariable temperature and conductivity RFID sensors for single-use biopharmaceutical manufacturing components

Single-use biopharmaceutical manufacturing requires monitoring of critical manufacturing parameters. We have developed an approach for passive radio-frequency identification (RFID)-based sensing that converts ubiquitous passive 13.56 MHz RFID tags into inductively coupled sensors. We combine several measured parameters from the resonant sensor antenna with multivariate data analysis and deliver unique capability of multiparameter sensing and rejection of environmental interferences with a single sensor. We demonstrate here the integration of these RFID sensors into single-use biopharmaceutical manufacturing components. We have tested these sensors for over 500 h for measurements of temperature and solution conductivity with the accuracy of 0.1°C (32-48°C range) and accuracy of 0.3-2.9 mS/cm (0.5-230 mS/cm range). We further demonstrate simultaneous temperature and conductivity measurements with an individual RFID sensor with the accuracy of 0.2°C (5-60°C range) and accuracy of 0.9 mS/cm (0.5-183 mS/cm range). Developed RFID sensors provide several important features previously unavailable from other single-use sensing technologies such as the same sensor platform for measurements of physical, chemical, and biological parameters; multi-parameter monitoring with individual sensors; and simultaneous digital identification.     

Effects of a Two‐Hour Change in Bedtime on the Sleep of Healthy Seniors

Some of the sleep disruption seen in seniors (>65 yrs) may be due to alteration of the circadian pacemaker phase and/or its phase angle with bedtime. The purpose of this study was to determine the effects of 2 h changes in the timing of bedtime (both earlier and later) on the sleep of seniors. Ten healthy seniors (9 F, 1 M, age 70–82 yrs) were each studied individually during three 120 h sessions (each separated by >2 weeks) in a time‐isolation laboratory. On nights 1 and 2, bedtime and rise‐time occurred at the subjects' habitual times; on nights 3–5, bedtime was specified by the experiment, but rise‐time was at the subjects' discretion (without knowledge of clock time). Under the control condition, subjects went to bed at their habitual bedtime (HBT), under the earlier bedtime condition at (HBT?2 h), and under the later bedtime condition at (HBT+2 h). Sleep was polysomnnographically recorded and rectal temperature continuously monitored. Although total sleep time increased in the earlier compared to the later condition (p<0.01), sleep efficiency decreased and wake after sleep onset increased (p<0.01). Subjective ratings of sleep were also worse under the earlier (HBT?2 h) than under later (HBT+ 2 h) condition (p<0.05). Performance did not differ between the earlier and later conditions. The larger the phase angle between actual bedtime and circadian temperature minimum (Tmin), the longer the time spent in bed and total sleep time, and the worse the sleep efficiency and subjective sleep ratings. There were no effects related to the phase angle between Tmin and rise‐time. The relative benefits of longer vs. more efficient sleep in the elderly require further investigation.     

Purification and characterization of a cellulase (CMCase) from a newly isolated thermophilic aerobic bacterium Caldibacillus cellulovorans gen. nov., sp. nov

One thermostable endoglucanase (CMCase) was purified to homogeneity from the culture supernatant of a new isolated thermophilic bacterium Caldibacillus cellulovorans. The molecular weight of the enzyme was 85.1 kDa as determined by SDS Polyacrylamide gel electrophoresis (PAGE) and 174 kDa by size-exclusion chromatography. The isoelectric point of the enzyme was at pH 4.12. The temperature for maximum activity was 80 °C, with half-lives of 32 min at 80 °C, and 2 min at 85 °C, and 83% activity remaining after 3 h at 70 °C. Thermostability of the enzyme was increased twofold by the addition of bovine serum albumin. Maximal activity was observed between pH 6.5 and 7.0. The enzyme activity was significantly inhibited by Zn²⁺, Hg²⁺, and p-chloromercuribenzenesulphonic acid. The enzyme showed high activity on carboxymethylcellulose (CMC) with much lower activity on Avicel; a low level of activity was also found against xylan. Cellobiose was the major product of hydrolysis of amorphous cellulose and CMC. Viscometric analysis indicated that the enzyme hydrolysed CMC in an exo-acting fashion. Cellotriose and cellobiose were not degraded and at least four contiguous glucosyl residues were necessary for degradation by the enzyme. The K _m and V _max of the enzyme for CMC were 3.4 mg ml^–1 and 44.7 mol min^–1 (mg protein)^–1, respectively.     

A likelihood ratio method for studying the re-entrainment of circadian rhythms

A simple technique is described that enables the degree of adaptation at different times of day to be studied in organisms that have been subjected to a sudden change in the Zeitgeber (as typically found in shiftwork or after transmeridian flight). The investigator is thus able to place greater emphasis on some readings than on others and to measure the amount of disruption at a particular time of day (e.g. for an important meeting) rather than to merely rely upon an overall daily measure. The method uses the likelihood ratio statistics which also allows a powerful test to be made of the statistical significance of any apparent adaptation. The method is illustrated on some 'jet-lag' data, where it was able to determine differential rates of adaptation according to the time of day at which the subjects were tested.