Statistical analysis of the micronucleus test--a modelling approach

This paper suggests an approach according to the principle that 'An experimental design is related to a statistical model describing that particular design'. A simple linear model approach is presented as an alternative to earlier proposed multiple single tests which is suitable for situations in which there are just a few dose levels and expression times. Furthermore, the aim is to move from significance testing towards a modelling and estimation procedure in order to find a structure and try to evaluate what is being determined in the experiment.     

水分胁迫对柑橘果皮细胞壁结构与代谢的影响

研究水分胁迫下,盆栽'暗柳橙(Citrus Sinensis Osbeck cv. Anliu)'的果实成熟期果皮细胞壁超微结构、细胞壁物质成分、细胞壁代谢相关酶活性的变化规律及其之间关系.结果表明,在果实发育成熟期,果皮细胞壁代谢相关水解酶果胶酶、纤维素酶、果胶甲酯酶的活性随着水分胁迫的加强而增加,多酚氧化酶活性与果胶酶活性变化趋势相反,果皮细胞壁代谢相关成分离子结合型果胶、共价结合型果胶、半纤维素、纤维素的含量随着水分胁迫的加强而降低,水溶性果胶含量随着水分胁迫的加强而增加,果皮细胞壁超微结构随着水分胁迫的加强而加速解体.     

A 'misplaced chapter' in the history of photosynthesis research; the second publication (1796) on plant processes by Dr Jan Ingen-Housz,MD, discoverer of photosynthesis

In 1779, the Dutch physician Jan Ingen-Housz (1730–1799) obtained a leave-of-absence from his post as Court Physician to Empress Maria Theresa of Austria in order to do research (in England) on plants during the summer months. He performed more than 500 experiments, and described the results in his exceptional book Experiments Upon Vegetables (1779). In addition to proving the requirement for light in photosynthesis, Ingen-Housz established that leaves were the primary sites of the photosynthetic process. Later, Ingen- Housz published research papers on various subjects but aside from his 1779 book, he published only one more communication on photosynthesis and plant physiology. This was entitled 'An Essay on the Food of Plants and the Renovation of Soils'. The essay was published in 1796 as an appendix to an obscure British government report, which is rare and virtually unknown. The present paper describes the 1796 essay, which is particularly interesting in that it shows how Ingen-Housz's concepts were modified by new interpretations of chemical phenomena described in Lavoisier's great and revolutionary book Traité Élémentaire de Chimie (1789). Ingen-Housz not only discovered photosynthesis, but plant respiration as well, and the 1796 essay is testimony to his remarkable insights.     

Nasal hyperreactivity and inflammation in allergic rhinitis

The history of allergic disease goes back to 1819, when Bostock described his own 'periodical affection of the eyes and chest', which he called 'summer catarrh'. Since they thought it was produced by the effluvium of new hay, this condition was also called hay fever. Later, in 1873, Blackley established that pollen played an important role in the causation of hay fever. Nowadays, the definition of allergy is 'An untoward physiologic event mediated by a variety of different immunologic reactions'. In this review, the term allergy will be restricted to the IgE-dependent reactions. The most important clinical manifestations of IgE-dependent reactions are allergic conjunctivitis, allergic rhinitis, allergic asthma and atopic dermatitis. However, this review will be restricted to allergic rhinitis. The histopathological features of allergic inflammation involve an increase in blood flow and vascular permeability, leading to plasma exudation and the formation of oedema. In addition, a cascade of events occurs which involves a variety of inflammatory cells. These inflammatory cells migrate under the influence of chemotactic agents to the site of injury and induce the process of repair. Several types of inflammatory cells have been implicated in the pathogenesis of allergic rhinitis. After specific or nonspecific stimuli, inflammatory mediators are generated from cells normally found in the nose, such as mast cells, antigen-presenting cells and epithelial cells (primary effector cells) and from cells recruited into the nose, such as basophils, eosinophils, lymphocytes, platelets and neutrophils (secondary effector cells). This review describes the identification of each of the inflammatory cells and their mediators which play a role in the perennial allergic processes in the nose of rhinitis patients.     

Structural requirements of (2'-5') oligoadenylate for protein synthesis inhibition in human fibroblasts.

The structural requirements of (2'-5')-oligoadenylic acid (pppA(2'p5'A)x, X greater than or equal to 1 or (2'-5'An) for inhibition of protein synthesis in cells were examined with a modified calcium-coprecipitation technique, using a series of trinucleotide analogs (pppA2'p5'A2'p5'N, N=rC, rG, rU, T, dC, dG, dA). In this system both the degree and the duration of the inhibition of protein synthesis were dependent on the added concentration of (2'-5')A3. Of all the heterotrimers, only the deoxy A derivative was active as an inhibitor of protein synthesis, while the other members of the analog series were found to have no inhibitory effects. In competition experiments between (2'-5')A3 and the non-active analogs, three heterotrimers were shown to reduce the activity of (2'-5')A3 in protein inhibition. In contrast, the dephosphorylated (2'-5')A3 had no inhibitory effect and was not effective in blocking (2'-5')A3. These results indicate that the 5'-terminal triphosphate is important for binding of (2'-5')A3 to the site of (2'-5')An action and the adenine base at the 2'-terminus is important for activating the machinery responsible for protein synthesis inhibition in the cells, most likely the (2'-5')An-activated nuclease.     

石灰岩珍稀濒危植物对酸性土壤的迁地适应性

通过半定量评价法,对81种石灰岩珍稀濒危植物迁移到桂林植物园酸性土壤上的适应性进行了研究。结果表明:适应的占54%,基本适应的占27%,欠适应的占11%,不适应的占8%。引种的大部分石灰岩珍稀濒危植物能够适应或基本适应桂林植物园的生境条件,而在不能较好地适应迁移地生境的物种中,小部分是由于土壤因素的影响,其余则是因气候条件的差异所致。     

Adaptive response for chromosomal inversions in pKZ1 mouse prostate induced by low doses of X radiation delivered after a high dose

Adaptive responses are induced by stress such as X radiation and result in a lower than expected biological response. Two-dose adaptive response experiments typically involve a low priming dose followed by a subsequent high radiation dose. Here, we used a sensitive in vivo chromosomal inversion assay to demonstrate for the first time an adaptive response when a low dose (0.01-1 mGy) was given several hours after a high 1000-mGy radiation dose. The adaptive responses in this study were of similar magnitude to the two-dose adaptive responses previously observed in this test system when the low dose was given first. A chromosomal inversion adaptive response was also induced by two 1000-mGy doses and when a 1-mGy dose was preceded or followed by a dose of 0.01 mGy, but not by two 4000-mGy doses. This is also the first example of an adaptive response when both doses are low. Our data agree with previous reports of an on-off mechanism of adaptive response. The induction of an adaptive response by a low dose after a high damaging dose provides evidence that the mechanisms underlying radiation adaptive responses are not due to prevention of damage induced by the high dose but to modulation of the cellular response to this damage.     

Basal metabolic rate and the evolution of the adaptive immune system

Vertebrates have evolved an adaptive immune system in addition to the ancestral innate immune system. It is often assumed that a trade-off between costs and benefits of defence governs the evolution of immunological defence, but the costs and benefits specific to the adaptive immune system are poorly known. We used genetically engineered mice lacking lymphocytes (i.e. mice without adaptive, but with innate, immunity) as a model of the ancestral state in the evolution of the vertebrate immune system. To investigate if the magnitude of adaptive defence is constrained by the energetic costs of producing lymphocytes etc., we compared the basal metabolic rate of normal and lymphocyte-deficient mice. We found that lymphocyte-deficient mice had a higher basal metabolic rate than normal mice with both innate and adaptive immune defence. This suggests that the evolution of the adaptive immune system has not been constrained by energetic costs. Rather, it should have been favoured by the energy savings associated with a combination of innate and adaptive immune defence.     

胃粘膜血流量对大鼠胃粘膜适应性细胞保护作用的影响

采用氢气清除法测定胃粘膜血流量,观察大鼠胃内灌洗低浓度盐酸酒精后灌注高浓度盐酸酒精（０．６ｍｏｌ／ＬＨＣｌ＋１５％ＥｔＯＨ）对ＧＭＢＦ和胃粘膜损害的影响。以及ＧＭＢＦ在适应性细胞保护中的作用。结果如下：（１）先用低浓度盐酸酒精作为弱刺激灌注随之以高浓度盐酸酒精作为强刺激灌注,引起胃粘膜适应性保护现象,它表现为：大体操作和损伤深度分别比单独泊组减少４７．０９％和４４．５７％应性现象,它表现为：大体损     

Kinetics of the Early Adaptive Response to Gamma Rays: Induction of a Cellular Radioprotective Mechanism in Murine Leukocytes <Emphasis Type="Italic">In vivo</Emphasis>

The aims of the study were to establish the kinetics of the early adaptive response and to determine the minimum adaptive dose of gamma rays capable of inducing this response. The minimum adaptive dose was determined by exposing groups of five BALB/c male mice to an adaptive dose of 0.005 or of 0.02 Gy γ rays from a ¹³⁷Cs source and challenge with 1.0 Gy 60 min later. The kinetics of adaptive response induction was established by exposing mice to an adaptive dose of 0.01 Gy, and subsequently to a challenge dose of 1.0 Gy at different times. Blood samples were collected from the tail immediately after exposure to the challenge dose, and the percentage of DNA-damaged cells and the extent of damaged were determined by single cell gel electrophoresis in 300 leukocytes per animal in five mice. The results confirms the capability of an in vivo induction of an early radioprotective process against the DNA-damage produced by gamma rays in murine leukocytes, and allows us to conclude that the minimum adaptive dose lies between 0.005 and 0.01Gy of gamma rays, and the early adaptive response is induced as early as 30 min after the exposure and persists for at least 18 hr.     

Was Wright Right? The Canonical Genetic Code is an Empirical Example of an Adaptive Peak in Nature; Deviant Genetic Codes Evolved Using Adaptive Bridges

The canonical genetic code is on a sub-optimal adaptive peak with respect to its ability to minimize errors, and is close to, but not quite, optimal. This is demonstrated by the near-total adjacency of synonymous codons, the similarity of adjacent codons, and comparisons of frequency of amino acid usage with number of codons in the code for each amino acid. As a rare empirical example of an adaptive peak in nature, it shows adaptive peaks are real, not merely theoretical. The evolution of deviant genetic codes illustrates how populations move from a lower to a higher adaptive peak. This is done by the use of “adaptive bridges,” neutral pathways that cross over maladaptive valleys by virtue of masking of the phenotypic expression of some maladaptive aspects in the genotype. This appears to be the general mechanism by which populations travel from one adaptive peak to another. There are multiple routes a population can follow to cross from one adaptive peak to another. These routes vary in the probability that they will be used, and this probability is determined by the number and nature of the mutations that happen along each of the routes. A modification of the depiction of adaptive landscapes showing genetic distances and probabilities of travel along their multiple possible routes would throw light on this important concept.     

A consultant's perspective on the regulatory hurdles to adaptive trials

This is a discussion of the following two papers appearing in this special issue on adaptive designs: 'A regulatory view on adaptive/flexible clinical trial design' by H. M. James Hung, Robert T. O'Neill, Sue-Jane Wang and John Lawrence and 'Confirmatory clinical trials with an adaptive design' by Armin Koch.     

Effect of endogenous carotenoids on "adaptive" mutation in Escherichia coli FC40

The appearance over many days of Lac(+) frameshift mutations in Escherichia coli strain FC40 incubated on lactose selection plates is a classic example of apparent "adaptive" mutation in an episomal gene. We show that endogenously overproduced carotenoids reduce adaptive mutation under selective conditions by a factor of around two. Carotenoids are known to scavenge singlet oxygen suggesting that the accumulation of oxidative base damage may be an integral part of the adaptive mutation phenomenon. If so, the lesion cannot be 7,8-dihydro-8-oxoguanine since adaptive mutation in FC40 is unaffected by mutM and mutY mutations. If active oxygen species such as singlet oxygen are involved in adaptive mutation then they should also induce frameshift mutations in FC40 under non-selective conditions. We show that such mutations can be induced under non-selective conditions by protoporphyrin photosensitisation and that this photodynamic induction is reduced by a factor of just over two when endogenous carotenoids are present. We argue that the involvement of oxidative damage would in no way be inconsistent with current understanding of the mechanism of adaptive mutation and the role of DNA polymerases.     

SOS mutator DNA polymerase IV functions in adaptive mutation and not adaptive amplification.

Adaptive point mutation and amplification are induced responses to environmental stress, promoting genetic changes that can enhance survival. A specialized adaptive mutation mechanism has been documented in one Escherichia coli assay, but its enzymatic basis remained unclear. We report that the SOS-inducible, error-prone DNA polymerase (pol) IV, encoded by dinB, is required for adaptive point mutation in the E. coli lac operon. A nonpolar dinB mutation reduces adaptive mutation frequencies by 85% but does not affect adaptive amplification, growth-dependent mutation, or survival after oxidative or UV damage. We show that pol IV, together with the major replicase, pol III, can account for all adaptive point mutations at lac. The results identify a role for pol IV in inducible genetic change.     

Adaptive designs from the viewpoint of an academic biostatistician

This is a discussion of the following three papers appearing in this special issue on adaptive designs: 'FDA's critical path initiative: A perspective on contributions of biostatistics' by Robert T. O'Neill, 'A regulatory view on adaptive/flexible clinical trial design' by H. M. James Hung, Robert T. O'Neill, Sue-Jane Wang and John Lawrence; and 'Confirmatory clinical trials with an adaptive design' by Armin Koch.     

Adaptive response in different mitotic cycles after irradiation

The frequency of cells with chromosome aberrations and the number of aberrations per cell have been studied by metaphase analysis in the nonirradiated progeny of irradiated human blood lymphocytes. DNA fragmentation (DNA double-stranded breaks) has been investigated by DNA comet assay. To study the adaptive response (AR), PHA-stimulated lymphocytes were irradiated by the adaptive dose (0.05 Gy) in 24 h and by challenge dose (1 Gy) in 48 h after stimulation. The first through fourth mitoses were identified by 5-bromodeoxyuridine. It was found that the frequency of chromosome aberrations and double-strand breaks were increased in all mitotic cycles after the challenge irradiation. In most individuals, the adaptive response is induced by adaptive and challenge irradiations in the first and the second mitotic cycles (48 and 72 h after stimulation, respectively); however, it is absent in the third and the fourth mitoses. In the first mitosis (1Gy in 48 h after stimulation), only chromatid aberrations are observed; chromosome aberrations were registered in subsequent mitoses. DNA comet assay showed that the adaptive response was obvious at 48–72 h, but not 96 h, after stimulation. It can be concluded that the nonirradiated progeny of irradiated lymphocytes have genomic instability. The adaptive response is manifested up to the third mitosis and is explained by the decreasing number of chromatid and chromosome aberrations and DNA fragmentation. We suppose that double-stranded DNA breaks may be damage signals for the induction of adaptive response.     

Methodological developments vs. regulatory requirements

This is a discussion of the following three papers appearing in this special issue on adaptive designs: 'FDA's critical path initiative: A perspective on contributions of biostatistics' by Robert T. O'Neill, 'A regulatory view on adaptive/flexible clinical trial design' by H. M. James Hung, Robert T. O'Neill, Sue-Jane Wang and John Lawrence; and 'Confirmatory clinical trials with an adaptive design' by Armin Koch.     

Opening the adaptive toolbox

This is a discussion of the following three papers appearing in this special issue on adaptive designs: 'A regulatory view on adaptive/flexible clinical trial design' by H. M. James Hung, Robert T. O'Neill, Sue-Jane Wang and John Lawrence; 'Confirmatory clinical trials with an adaptive design' by Armin Koch; and 'FDA's critical path initiative: A perspective on contributions of biostatistics' by Robert T. O'Neill.     

The microbial experience of environmental phosphate fluctuations. An essay on the possibility of putting intentions into cell biochemistry

We present a model of microbial information processing that contains characteristic features of the phenomenon of physiological adaptation. The backbone of the model is the "adaptive event" in which energy-converting subsystems of the cell interact with the changing environment. In this process, the subsystems pass, via an adaptive operation mode, from one adapted state to the next. An adaptive operation mode takes place when an adapted state is disturbed by an environmental alteration. These two manifestations of an adaptive event were differently treated in the simulation, based on an application of linear irreversible thermodynamics to the energy transduction of adaptive subsystems. In adapted states, the conductivity coefficients of the flow-force relationships employed remained constant, whereas during an adaptive operation mode, these coefficients were altered in a directional manner during the simulation. An example dealing with the complex relationship between phosphate uptake and cyanobacterial growth is given. In this example, the simulation of adapted states of two subsystems of the incorporating machinery, namely the phosphate carrier in the cell membrane and the F-ATPase in the thylakoid membrane, was in accordance with the measured uptake kinetics, and when fixed, predetermined conductivity coefficients were used. In the adaptive operation mode, however, the simulated behavior was in agreement with experimental observations when the program was able to "interpret" its own performance in the light of environmental phosphate fluctuations, experienced by the cell in the past, and to reconstruct the two subsystems according to this interpretation. Via transitions between adapted states and adaptive modes, information is transferred from one adaptive event to the next: the latter "inherits" the results of former interpretations. By appropriating them selectively, it is entering into a future in which its own interpretation is passed on to the following adaptive event. The model is discussed with respect to the concept of autopoiesis.