Comparative Ploidy Studies Using Cytological and Paraffin Section Preparations

The ploidy profiles of benign and malignant tumours can be obtained using image analysis. However, the results of ploidy studies have varied according to the type of specimen used. We compared the ploidy profiles of paraffin embedded thin sections, cytospin preparations of disaggregated cells, and cytological smears from the same specimen as defined by image analysis. Ten benign breast lesions, 10 breast carcinomas and 10 malignant melanomas were investigated in this way. Preparations stained by the Feulgen technique were examined using the MD20 video image analysis densitometry system. Ploidy profiles were obtained by measuring the integrated optical density of at least 200 nuclei. By paying proper attention to the quality of fixation and presence or absence of cytoplasm around cells, comparable results were found for all preparations in each case. We therefore conclude that if careful attention is paid to the technical quality of the material, reliable ploidy results can be obtained by image analysis.     

Effect of pea bacterial blight (Pseudomonas syringae pv. pisi) on the yield of spring sown combining peas (Pisum sativum)

Field trials to examine the effect of pea bacterial blight (Pseudomonas syringae pv. pisi) (Psp) on the yield of combining peas were carried out at five sites (HRI Wellesbourne, ADAS Rosemaund, ADAS Terrington, PGRO, SASA East Craigs) in the UK in 1990, 1991 and 1992. Healthy seed, cv. Solara, and seed naturally infected with Psp Race 2 was sown in large plots (c. 200 m²) in the open or under nets to prevent bird damage by pigeons. Despite relatively low disease severity levels (< 15% leaf area) and separation by at least 12 m of cv. Consort (resistant to Race 2) between plots there was considerable spread of disease into plots sown with healthy seed. Regression analysis demonstrated a significant relationship between yield and disease. Of the disease measurements examined, disease severity on the leaves (stipules) at growth stage 208 was found to be the best predictor of yield. A model containing environmental and bird damage terms, in addition to disease, suggested that a yield loss of 0.98 t/ha would be expected for a disease severity score of 1, equivalent to 5% leaf area affected, at growth stage 208.     

Energy coupling sites in the electron transport chain of Zymomonas mobilis

Abstract Energy-coupling sites in the electron transport chain of the obligately fermentative aerotolerant bacterium Zymomonas mobilis were examined. The H⁺ /O stoichiometry of the electron transport chain in intact bacteria oxidizing ethanol was close to 3.3. Cytoplasmic membrane vesicles coupled NADH oxidation to ATP synthesis. With ascorbate/phenazine methosulfate they showed oxygen uptake which was sensitive to antimycin A, but no significant ATP synthesis could be detected. Cells with a defective coupling site I, prepared by cultivation on a sulfate-deficient medium, showed a decreased rotenone sensitivity of respiration, and they lacked almost all the respiration-driven proton translocation and ATP synthesis. We conclude that, despite the reported composition of the electron transport chain, only energy coupling site 1 was functional in Z. mobilis .     

Determination of adenosine triphosphate in human semen to estimate the fertilizing potential and to quantify sperm antibodies

The measurement of the ATP content of fresh semen is as accurate as the estimation of sperm motility by conventional methods in discriminating between semen of fertile versus subfertile men. The ATP content of frozen thawed donor semen is correlated with the probability of conception per cycle of insemination. Exact quantification of cytotoxic sperm antibodies in serum is possible with the adenosine-triphosphate-release-cytotoxicity test, since measurement is free of the bias of microscopic examination. The procedure has been simplified by testing only one serum dilution and calculating the ‘sperm toxicity index’.     

Some New Aspects of 2 × 2 Table Measures and Their Applications

The paper deals with the analysis of 2 × 2 table measures and describes several new possibilities for applying them. An estimate is given for HELLMICH's asymmetric K-measure and a possible extension of the contingency concept in a special 2×2 case is presented. The concept introduced and the recommended computer application of the 2×2 table measures is supported by new results obtained for the relationship between jaw joint pains and noises examined earlier by F. SCHMID and C. ZSCHEGE. With a larger number of observations the opportunity arises for a deeper analysis of this dental problem using path analysis. The plan of evaluation and the conncetion between the variables examined is illustrated by a path-diagram. The association tables determined on the basis of the (C) values introduced during the examination of jaw joint disturbances using association analysis draw attention to the special role of the left side of mastication. The paper raises ideas for further generalisations and indicates where further research is needed.     

The effects of X-rays on the proliferation dynamics of cells in the imaginal wing disc ofDrosophila melanogaster

Summary We report on the size distribution of clones marked by mitotic recombination induced by several different doses of X-rays applied to 72 h oldDrosophila larvae. The results indicate that the radiation significantly reduces the number of cells which undergo normal proliferation in the imaginal wing disc. We estimate that 1000 r reduces by 40–60% the number of cells capable of making a normal contribution to the development of the adult wing. Part of this reduction is due to severe curtailment in the proliferative ability of cells which nevertheless remain capable of adult differentiation; this effect is possibly due to radiation-induced aneuploidy. Cytological evidence suggests that immediate cell death also occurs as a result of radiation doses as low as 100 r. The surviving cells are stimulated to undergo additional proliferation in response to the X-ray damage so that the result is the differentiation of a normal wing.     

Phylogeography of the leaf beetle Chrysolina virgata in wetlands of Japan inferred from the distribution of mitochondrial haplotypes

The genetic differentiation among populations of the leaf beetle Chrysolina virgata living in wetlands of Japan was studied based on the sequence data of the mitochondrial cytochrome oxidase subunit I gene region (750 bp). Two distinct lineages of mitochondrial haplotypes were found: one (clade A) consisted of 26 haplotypes distributed over the distribution range of C. virgata between north‐east Honshu and Kyushu, whereas the other (clade B) was monotypic and confined to a small region in north‐east Honshu where it coexisted with clade A. Nested clade analysis for these haplotypes suggested that range expansion and following differentiation due to isolation by distance might have resulted in the present distribution pattern of the haplotypes in clade A. We discuss the evolutionary process leading to the occurrence of two distinct haplotype clades in Japan in terms of repeated colonization from the continent and range expansion and contraction during climatic changes.     

Conditional Maximum Likelihood Estimation Following a Group Sequential Test

We consider estimation after a group sequential test. An estimator that is unbiased or has small bias may have substantial conditional bias (Troendle and Yu, 1999, Coburger and Wassmer, 2001). In this paper we derive the conditional maximum likelihood estimators of both the primary parameter and a secondary parameter, and investigate their properties within a conditional inference framework. The method applies to both the usual and adaptive group sequential test designs. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Integrated palaeoecological and historical data in the service of fine-resolution land use and ecological change assessment during the last 1000 years in Rõuge, southern Estonia

Aims Our aim is to reconstruct decadal scale development of historical landscapes during the last 1000 years by means of fossil pollen analysis of annually laminated lake sediments, and detailed historical maps and documents. Location Lake Rõuge Tõugjärv (Estonia), a small lake with annually laminated lake sediments situated in a dense prehistoric setting. Methods The chronology of the palaeodata is based on the annual laminations supported by AMS ¹⁴C and ²¹⁰Pb dating and ¹³⁷Cs, ²⁴¹Am, and spheroidal carbonaceous particle marker horizons. The time‐scale and resolution allows fine sampling (the pollen samples generally comprise 3.5 years) and vegetation change reconstruction. Relevant source area of pollen (RSAP) of the lake was estimated, and the statistical zonation, rate of change, palynological richness, and DCA and PCA ordinations were generated on the basis of the pollen data. The historical calibration data set (maps, numerical information on population, domestic stock, farmland division, etc.) is based on archival material preserved in the Estonian Historical Archives. Results The topmost part (0–180 cm) of the sediment column of Lake Rõuge Tõugjärv, covering the last 1000 years, is visibly laminated carbonaceous gyttja. The varve chronology extends from ad 2000 to ad 1339, with a cumulative ± 9‐year error estimate. Beyond this the chronology is extrapolated using the ¹⁴C date and varve age–depth estimations. The simulation of the RSAP of Lake Tõugjärv shows that the major portion of the pollen loading originating from local vegetation is derived from plants growing within 2000 m of the sampling site. The pollen record divides into five statistically significant subgroups, which fall on the PCA plot into three clusters reflecting the general openness–closedness of the landscape. During the period between ad 1000 and 1200 (RT 1) the Rõuge area was generally wooded with birch, spruce and pine forests. The advancement of extensive farming gradually opened up the landscape between ad 1200 and 1650 (RT 2 and RT 3). The maximum openness of the landscape was reached between ad 1650 and 1875 (RT 4), with the most open period in the late eighteenth century. Historical maps from 1684 and 1870–99 and available quantitative data on population, domestic stock, farmland division, etc. show the same trend. The pollen data covering the last 125 years, and maps from 1935 and 1995, show the reduction of arable land in RSAP of the lake under investigation and the reduction of open land to an extent comparable with the end of the seventeenth century. Main conclusions The formation and development of the cultural landscape at Rõuge over the last 1000 years is characterized by rapid changes in floristic richness and rates of vegetation change attributed to certain historic processes in the RSAP. Five phases of landscape and social development are clearly distinguished during the last 1000 years. The decadal scale vegetation response to human‐induced forcing agrees with historical maps and documents and could be used for past landscapes prior to the period with solid historical data.