Cellular senescence, rejuvenation and potential immortality.

Ageing, death, and potential immortality lie at the heart of biology, but two seemingly incompatible paradigms coexist in different research communities and have done since the nineteenth century. The universal senescence paradigm sees senescence as inevitable in all cells. Damage accumulates. The potential immortality paradigm sees some cells as potentially immortal, especially unicellular organisms, germ cells and cancerous cells. Recent research with animal cells, yeasts and bacteria show that damaged cell constituents do in fact build up, but can be diluted by growth and cell division, especially by asymmetric cell division. By contrast, mammalian embryonic stem cells and many cancerous and 'immortalized' cell lines divide symmetrically, and yet replicate indefinitely. How do they acquire their potential immortality? I suggest they are rejuvenated by excreting damaged cell constituents in extracellular vesicles. If so, our understanding of cellular senescence, rejuvenation and potential immortality could be brought together in a new synthesis, which I call the cellular rejuvenation hypothesis: damaged cell constituents build up in all cells, but cells can be rejuvenated either by growth and cell division or, in 'immortal' cell lines, by excreting damaged cell constituents. In electronic supplementary material, appendix, I outline nine ways in which this hypothesis could be tested.

The production of auxin by dying cells.

In this review, I discuss the possibility that dying cells produce much of the auxin in vascular plants. The natural auxin, indole-3-acetic acid (IAA), is derived from tryptophan by a two-step pathway via indole pyruvic acid. The first enzymes in the pathway, tryptophan aminotransferases, have a low affinity for tryptophan and break it down only when tryptophan levels rise far above normal intracellular concentrations. Such increases occur when tryptophan is released from proteins by hydrolytic enzymes as cells autolyse and die. Many sites of auxin production are in and around dying cells: in differentiating tracheary elements; in root cap cells; in nutritive tissues that break down in developing flowers and seeds; in senescent leaves; and in wounds. Living cells also produce auxin, such as those transformed genetically by the crown gall pathogen. IAA may first have served as an exogenous indicator of the presence of nutrient-rich decomposing organic matter, stimulating the production of rhizoids in bryophytes. As cell death was internalized in bryophytes and in vascular plants, IAA may have taken on a new role as an endogenous hormone.

Serum screening for Down syndrome and adverse pregnancy outcomes: a case-controlled study.

The relationship between adverse perinatal outcomes in women with false positive biochemical screening test for Down syndrome was investigated in a retrospective case-controlled study. A cohort of 4000 women who booked for routine antenatal care and opted for biochemical screening over a 22 month period was obtained. The pregnancy outcome data of 272 women with a false positive screening test for Down syndrome (risk >1 in 250) at 15-18 weeks of gestation (study group) were compared with data from 272 age and gestation matched controls with a negative Down syndrome screening test from the same population. The frequency of normal and adverse perinatal outcomes, including pre-eclampsia, isolated intrauterine growth restriction, spontaneous preterm labour and stillbirth was recorded. The incidence of adverse pregnancy outcomes was 11.9% in the study group and 8.6% in the control group. The estimated odds ratio of an abnormal outcome in the study group was 1.41 (95% CI-0.790, 2.55). The observed difference between proportion was 0.0324 (95% CI-0.022, 0.083; p=0.40). These data identify no evidence for a strong association between a false positive Down syndrome screening test result and subsequent adverse perinatal outcomes in the general population.

Perinatal outcome in pregnancies with a positive serum screening for Down's syndrome due to elevated levels of free beta-human chorionic gonadotropin.

OBJECTIVE: To evaluate the potential clinical use of maternal serum free beta-human chorionic gonadotropin (beta-hCG) and uterine artery Doppler investigation to screen for placenta-related adverse outcome in pregnancies at positive risk for Down's syndrome at 15-18 weeks. DESIGN: A cohort of 329 consecutive pregnant women with a singleton viable pregnancy and a positive risk for Down's syndrome was retrospectively investigated. This group was obtained from an unselected population of 3952 women attending the same hospital over a 2-year period. Using the results of this first analysis, we selected a group of 26 women with unexplained high levels of free beta-hCG and followed them prospectively with monthly ultrasound and uterine artery Doppler examinations. RESULTS: In the retrospective cohort, risk ratios stratified for maternal serum beta-hCG multiple of the median (MoM) values indicated that the highest incidence of adverse pregnancy outcome was in those women with values of > or = 5.0. In the prospective study, pregnancy outcome was complicated by uteroplacental disorders in eight cases. Analysis of the Doppler investigation indicated that, in women with a very high level of hCG, an abnormally high uterine artery pulsatility index (PI) had lower sensitivity and negative predictive value than early diastolic notch, whereas the specificity and positive predictive value were higher for a high uterine artery PI. CONCLUSIONS: These findings suggest an association between a high level of maternal serum beta-hCG at 15-18 weeks, the presence of an early diastolic notch in the uterine artery flow velocity waveform and adverse pregnancy outcome due to abnormal development of the uteroplacental circulation. Young women with an unexplained high beta-hCG level would benefit, apart from detailed sonography of the fetus and/or karyotyping, from uterine Doppler investigation and counselling about the follow-up and management of placenta-related pregnancy disorders.

The case for fan and pump motor speed control.

For many years buildings were designed to use standard plant and provide good operational conditions, without consideration of the efficient use of energy. This might have been acceptable when energy was cheap, but with rising energy costs, and recent government instruction to reduce consumption levels, it is necessary to actively consider plant running costs.

Effects of osmotic stress on polar auxin transport in Avena mesocotyl sections.

Segments of mesocotyls of Avena sativa L. transported [1-(14)C]indol-3yl-acetic acid (IAA) with strictly basipetal polarity. Treatment of the segments with solutions of sorbitol caused a striking increase in basipetal auxin transport, which was greatest at concentrations around 0.5 M. Similar effects were observed with mannitol or quebrachitol as osmotica, but with glucose or sucrose the increases were smaller. Polar transport was still detectable in segments treated with 1.2 M sorbitol. The effects of osmotic stress on the polar transport of auxin were reversible, but treatment with sorbital solutions more concentrated than 0.5 M reduced the subsequent ability of mesocotyl segments to grow in response to IAA. The increased transport of auxin in the osmotically stressed segments could not be explained in terms of an increased uptake from donor blocks. The velocity of transport declined with higher concentrations of osmoticum. The reasons for the enhancement of auxin transport by osmotic stress are not known.

Carrier-mediated auxin transport.

1. Auxin (IAA) transport was investigated using crown gall suspension tissue culture cells. We have shown that auxin can cross the plasmalemma both by transport of IAA anions on a saturable carrier and by passive (not carriermediated) diffusion of the lipid-soluble undissociated IAA molecules (pK=4.7). The pH optimum of the carrier for auxin influx is about pH 6 and it is half-saturated by auxin concentrations in the region of 1-5 µM. We found that the synthetic auxin 2,4D specifically inhibited carrier-mediated IAA anion influx, and possibly also efflux. Other lipid-soluble weak acids which are not auxins, such as 3,4-dichlorobenzoic acid, had no effect on auxin transport. By contrast, we found that TIBA, an inhibitor of polar auxin transport in intact tissues inhibited only the carrier-mediated efflux of IAA. 2. When the pH outside the cells is maintained below that of the cytoplasm (pH 7), auxin can be accumulated by the cells: In the initial phase of uptake, the direction of the auxin concentration gradient allows both passive carrier-mediated anion influx (inhibited by 2,4D) and a passive diffusion of undissociated acid molecules into the cells. Once inside the cytoplasm, the undissociated molecules ionise, producing IAA anions, to a greater extent than in the more acidic extracellular environment. Uptake by passive diffusion continues as long as the extracellular concentration of undissociated acid remains higher than its intra-cellular concentration. Thus, the direction of the auxin anion concentration gradient is reversed after a short period of uptake and auxin accumulates within the cells. The carrier is now able to mediate passive IAA anion efflux (inhibited by TIBA) down this concentration gradient even though net uptake still proceeds because the carrier is saturable whereas passive diffusion is not. 3. Auxin "secretion" from cells is regarded as a critical step in polar auxin transport. The evidence which we present is consistent with the view that auxin "secretion" depends on a passive carrier-mediated efflux of auxin anions which accumulate within the cells when the extra-cellular pH is below that of the cytoplasm. The implications of this view for theories of polar auxin transport are discussed.

Cellulase and cell differentiation in Acer pseudoplatanus.

Homogenates of differentiating xylem and phloem tissue have higher cellulase activities than cambial samples; the highest activity is always found in phloem. Callus tissue, in which no vascular differentiation occurs, contains only low cellulase activity. The results suggest that cellulase is involved in vascular differentiation. Different pH optima of cellulase activity were found: in cambium, xylem and phloem tissue, cellulase activity with an optimum at about pH 5.9 is predominantly membrane-bound; it is sedimentable at 100,000 g and releasable by Triton X-100. The same may be true of activity with an optimum at pH 5.3. Phloem tissue also contains a soluble, cytoplasmic cellulase of high activity at pH 7.1, and xylem tissue contains cytoplasmic cellulase with an optimum at pH 6.5. Low cellulase activity with a pH optimum similar to that of xylem homogenates was found in xylem sap. Cellulase activity in abscission zones increases greatly just before leaf abscission. Abscission zone cellulase has two pH optima, et 5.3 and 5.9; both activities are increased by Triton treatment of homogenates. The possible existence of several different cellulases forming part of a cellulase complex, and the rôle of the enzymes in hydrolysing wall material during cell differentiation are discussed.

Cellulase in latex and its possible significance in cell differentiation.

Cellulase was found to be present in the latex of species with articulated laticifers but it could not be detected in the latex of species with nonarticulated laticifers. It is suggested that cellulase is involved in the removal of end walls during the differentiation of articulated laticifers.