Effect of 2,4-D and isoproturon on chromosomal disturbances during mitotic division in root tip cells of <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

The widespread use of the herbicides for weed control and crop productivity in modern agriculture exert a threat on economically important crops by way of cytological damage to the cells of the crop plant or side effects, if any, induced by the herbicides. In the present communication, author describes the effects of 2,4-D and fsoproturon on chromosomal morphology in mitotic cells of Trittcum aestivum L. The wheat seedlings were treated with range of concentrations (50–1200 ppm) of 2,4-D and Isoproturon for 72 h at room temperature. In the mitotic cells, twelve distinct chromosome structure abnormalities were observed over control. The observed irregularities were stickiness, c-mitosis, multipolar chromosomes with or without spindles, fragments and bridges, lagging chromosomes, unequal distribution of chromosomes, over contracted chromosomes, unoriented chromosomes, star shaped arrangement of the chromosomes, increased cell size and failure of cell plate formation. The abnormalities like stickiness, fragments, bridges, lagging or dysjunction, unequal distribution and over contracted chromosomes meetfrequently.     

Activated charcoal filter effectively reduces p-benzosemiquinone from the mainstream cigarette smoke and prevents emphysema

In this paper, we have made a comparative evaluation of the cytotoxicity and pathophysiological effects of mainstream smoke from cellulose acetate (CA)-filtered cigarettes with that of charcoal-filtered cigarettes developed in our laboratory. Previously, we had demonstrated that the mainstream smoke from an Indian CA-filtered commercial cigarette contains p-benzosemiquinone (p-BSQ), a major, highly toxic, long-lived water-soluble radical. Here, we have examined 16 brands of different CA-filtered cigarettes including Kentucky research cigarettes, and observed that mainstream smoke from all the cigarettes contains substantial amounts of p-BSQ (100–200 μg/cigarette). We also show that when the CA filter is replaced by a charcoal filter, the amount of p-BSQ in the mainstream smoke is reduced by 73–80%, which is accompanied by a reduction of carbonyl formation in bovine serum albumin to the extent of 70–90%. The charcoal filter also prevented cytotoxicity in A549 cells as evidenced by MTT assay, apoptosis as evidenced by FACS analysis, TUNEL assay, overexpression of Bax, activation of p53 and caspase 3, as well as emphysematous lung damage in a guinea pig model as seen by histology and morphometric analysis. The results indicate that the charcoal filter developed in our laboratory may protect smokers from cigarette smoke-induced cytotoxity, protein modification, apoptosis and emphysema.     

The phase contrast microscope in plant cytology

Phase contrast microscopy affords a rapid and easy way of counting chromosomes from root-tip cells of Angiosperms. The staining method is a modified Feulgen process, the chromosomes being understained before examination under phase. This cuts down even further the time for obtaining satisfactory chromosome counts. Under phase and using a green filter the understained chromosomes in squashes, after only 30 minutes in Feulgen, appear jet black within an almost transparent cell. There is no interference from the cellulose cell walls.     

Use of nuclepore filters for counting bacteria by fluorescence microscopy.

Polycarbonate Nuclepore filters are better than cellulose filters for the direct counting of bacteria because they have uniform pore size and a flat surface that retains all of the bacteria on top of the filter. Although cellulose filters also retain all of the bacteria, many are trapped inside the filter where they cannot be counted. Before use, the Nuclepore filters must be dyed with irgalan black to eliminate autofluorescence. Direct counts of bacteria in lake and ocean waters are twice as high with Nuclepore filters as with cellulose filters.     

Use of nuclepore filters for counting bacteria by fluorescence microscopy.

Polycarbonate Nuclepore filters are better than cellulose filters for the direct counting of bacteria because they have uniform pore size and a flat surface that retains all of the bacteria on top of the filter. Although cellulose filters also retain all of the bacteria, many are trapped inside the filter where they cannot be counted. Before use, the Nuclepore filters must be dyed with irgalan black to eliminate autofluorescence. Direct counts of bacteria in lake and ocean waters are twice as high with Nuclepore filters as with cellulose filters.     

The toxic effects of cadmium on cell division and chromosomal morphology of Hordeum vulgare

The effects of cadmium at different concentrations (0.5–20 ppm) on root growth, cell division and chromosomal morphology of Hordeum vulgare were studied. The rate of root growth and mitotic index decreased progressively with increasing cadmium concentration and treatment duration. Different concentrations of cadmium could cause mitotic irregularities comprising c-mitoses, anaphase bridges, breaks, stickiness, lagging and vagrant chromosomes and micronuclei. The intensity of the toxic effects is basically dependent on the cadmium concentration and duration of treatment.     

Involvement of knob heterochromatin in mitotic abnormalities in germinating aged seeds of maize.

Seeds of three inbred lines of maize, with contrasting numbers of heterochromatic knobs and stored under two different ageing treatments, were analyzed for the occurrence of abnormalities at mitotic anaphase in root meristems 3, 7, 21,42, and 56 days after germination, and in root meristems of their freshly harvested selfed progeny. The largest category of detectable aberrations involved breakage of knobbed chromosome arms. We have obtained evidence that knob heterochromatin plays a central role in the origin of primary chromosome bridges. The initial event responsible for the occurrence of breakages and lagging chromosomes was characterized by the nondisjunction of newly replicated sister chromatids, which was observed to occur preferentially at the knob level. Such configurations, and all the other types of abnormalities (as for example, lagging chromosomes, typical chromosome bridges, fragments, and micronuclei), were observed at decreasing frequencies throughout root growth. Nevertheless, we have detected the occurrence of breakage-fusion-bridge cycles that were initiated by broken chromosomes. The relationship between late-replicating DNA in maize knob heterochromatin and the vulnerability of such regions to breakage is discussed. Our observations suggest a similarity between the mechanisms involved or associated with the origin of the described abnormalities and those reported to occur in cultured maize cells.     

Cytotoxic effects of DON and T-2 toxin on plant cells

The effect of deoxynivalenol (DON) and T-2 toxin on mitotic index (MI) and relative division rate (RDR) in actively dividing onion (Allium cepa L.) root-tip cells was studied. Both these toxins resulted in decline of mitotic activity which was inversely proportional to the concentrations of these toxins. T-2 was more effective resulting in 59% RDR value at 2.5 ppm whereas DON treated root cells had 78% RDR at the same concentration as compared to respective sets of controls.     

Bronchial Reactivity to Cigarette and Cigar Smoke

The change in specific airway conductance produced by smoking a cigarette under standard conditions was measured in 91 heavy smokers. Subsequently 19 of the most reactive subjects smoked two cigarettes with different filters and another containing cigar tobacco. The results indicated that reactivity to cigarette smoke was reduced significantly by increasing the retention efficiency of the filter and that reactivity to inhaled cigar-tobacco smoke was no less than that to cigarette smoke.     

Transfer of a male-sterility-inducing cytoplasm from onion to leek ( Allium ampeloprasum)

Two interspecific triploid (AAC) hybrids (84/1-94 and 99/1-94) from crosses between onion [ Allium cepa (2 n=2 x=16, CC)] and leek [ A. ampeloprasum (2 n=4 x=32, AAAA)] were backcrossed to leek in order to transfer a male-sterility-inducing cytoplasm from onion that would enable the production of hybrid leek. GISH evaluations of meiosis in the interspecific hybrids revealed irregularities due to univalent onion chromosomes producing micronuclei from onion chromatin, whereas the pairing of the two sets of leek chromosomes was nearly normal. Attempts to use colchicine to double the chromosome number of the hybrids failed. Backcrosses of 84/1-94 to leek as the pollen parent were not successful. The first backcross of 99/1-94 to tetraploid leek produced 11 BC(1) plants with chromosome numbers between 38 and 41. Identification of parental chromosomes by GISH showed that all eight onion chromosomes and 30-33 leek chromosomes were transmitted to the backcross progenies due to unreduced egg cells. Onion chromosomes were eliminated during the second backcross. Southern hybridization confirmed the transfer of the T-cytoplasm like source of CMS from onion to the BC(2) progenies. After the third backcross to leek, 158 plants were obtained with varying numbers of onion chromosomes and some intergenomic recombinant chromosomes. Alloplasmic leek plants without onion chromatin were selected for further characterization of male sterility and quality traits.     

Classification of chromosome segregation errors in cancer

Abnormal chromosome segregation at mitosis is one way by which neoplastic cells accumulate the many genetic abnormalities required for tumour development. In this paper, a straightforward morphology-based classification of chromosome segregation errors in cancer is suggested. This classification distinguishes between abnormalities in spindle symmetry (spindle multipolarity, size-asymmetry of ana-telophase poles) and abnormalities in sister chromatid segregation (chromosome bridges, chromatid bridges, chromosome lagging, acentric fragment lagging). Often, these categories of errors must be combined to accurately describe the events in a single abnormal mitotic cell. The suggested categories can to some extent be distinguished by standard chromatin staining. However, labelling of abnormal mitotic figures by fluorescence in situ hybridization and immunofluorescence enhances the accuracy of classification and also allows visualisation of the segregation of individual chromosomes, making it possible to detect non-disjunction also in the absence of gross alterations in mitotic morphology. Further characterisation of the molecular alterations leading to abnormal chromosome segregation together with the current developments in nano-level and real-time imaging will undoubtedly lead to an improved understanding of chromosome dynamics in cancer cells. Any morphology-based classification of chromosome segregation errors in cancer must therefore be taken as provisional, anticipating a satisfactory integration of morphology and molecular biology.     

Membrane filters: interference with liquid scintillation counting by the use of tissue solubilizers with cellulose ester membrane filters

Data are presented that show that the mixed esters of cellulose membrane filters, when treated with certain tissue solubilizers, produce a color quench which prevents adequate counting of radioactive samples. This color quench does not occur when using cellulose acetate membrane filters, or when two commercial liquids which have a tissue solubilizer precombined with the scintillation fluid are used with either type of filter. The data also show that the color reacton is not dependent upon the detergent used in these filters by the manufacturer.     

Effects of benzyladenine on prokaryotic and eukaryotic cells.

Comparative studies of the effect of benzyladenine (BA) on the yeast Saccharomyces cerevisiae, the bacterium Salmonella typhimurium, the shallot Allium ascalonicum and Chinese hamster fibroblast cells were performed. The tested substance had no mutagenic activity on yeast, bacteria and cultured fibroblast cells. Changes in mitotic activity and cell division abnormalities were observed after BA treatment in shallot root-tip cells.     

来自山羊草的杀配子染色体(Aegilops cylindryca 2C)对小黑麦减数分裂及花粉粒有丝分裂过程的影响

用来自柱穗山羊草的杀配子染色体2C,诱导六倍体、八倍体小黑麦染色体的断裂,观察杂种F1的减数分裂行为,在PMCI及PMCII后期观察到了大量的落后染色体、染色体断片、环状染色体及桥,在二分了解子及四分孢子中有为数甚多的、大小不一的微核,有的还形成多分孢子。在对F1花粉粒的有丝分裂观察中,未见分裂异常。由此推断杀配子染以体诱导染色体断裂可能不发生在配子形成的有丝分裂过程。这与巳有的报道不同。     

In vitro exposure of adenovirus types 5 and 7 in oropharyngeal secretions to cigarette smoke.

Adenovirus types 5 and 7 were suspended in clarified oropharyngeal secretions. After 1 ml of suspension was dispersed in a thin layer over a 25-cm2 surface in a flask, the suspension was exposed at 37 degrees C to eight 25-ml puffs of smoke from one cigarette. A mechanical smoking apparatus was used. Nonfilter cigarettes used had 23 mg of tar and 1.4 mg of nicotine, and filter cigarettes used had 19 mg of tar and 1.2 mg of nicotine. Smoke was flushed from the flask with normal filtered air. At 0, 0.25, and 1 h after exposure to smoke, untreated and smoke-treated viruses were titrated with monolayer cultures of human epithelioid (HEp-2) cells. Normal air was in contact with the suspensions between puffs and between smoke treatments and virus titrations. Smoke from filter or nonfilter cigarettes had no effect on the infectivity and replication of adenovirus types 5 and 7. Smoke from four cigarettes administered over a 4-h period caused a 2- to 3-log10 drop in the titers of both viruses. Smoke from four cigarettes was also highly toxic to HEp-2 host cells of the viruses.     

Polymerase chain reaction-gene probe detection of microorganisms by using filter-concentrated samples.

To detect low levels of microorganisms in environmental samples by using polymerase chain reaction (PCR)-gene probe detection, samples were concentrated by filtration. Fluoropore (Millipore Corp.) filters were compatible with PCR DNA amplification, whereas various other filters including nitrocellulose and cellulose acetate filters inhibited PCR amplification. By concentrating cells on Fluoropore filters and releasing the DNA by freeze-thaw cycling, PCR DNA amplification could be performed without removing the filter. Concentration with Fluoropore FHLP and FGLP filters permitted the detection of single cells of microorganisms in 100-ml samples by PCR-gene probes.     

Cytogenetic study of certain Tilia species]

The number of chromosomes, and mitosis and microsporogenesis were studied in Tilia europaea, T. tomentosa, T. cordata and T. platyphyllos under natural conditions of Voronezh region. Significant mixoploidy of meristematic tissue cells in seedling root tips, mitotic activity and meiosis have been demonstrated for the first time. The number of chromosomes in somatic cells varied from 28 to 140-146. The number 2n = 82 was modal for all the studied species. The number of chromosomes in T. europaea has been determined for the first time. The numbers of chromosomes in the genus Tilia are analysed on the basis of the literature and our own data. The peak of mitotic activity in seedlings and juvenile leaves (8-10%) fell on the night: 0-1 a.m. according to the winter time. Microsporogenesis is simultaneous and asynchronous. In some individual trees various abnormalities of meiosis were found: lagging in time in 1 to 5-6 chromosomes, bridges, fusions of spindles. Variations in pollen dimensions, shape and pore number were detected in many trees.     

Polymerase chain reaction-gene probe detection of microorganisms by using filter-concentrated samples.

To detect low levels of microorganisms in environmental samples by using polymerase chain reaction (PCR)-gene probe detection, samples were concentrated by filtration. Fluoropore (Millipore Corp.) filters were compatible with PCR DNA amplification, whereas various other filters including nitrocellulose and cellulose acetate filters inhibited PCR amplification. By concentrating cells on Fluoropore filters and releasing the DNA by freeze-thaw cycling, PCR DNA amplification could be performed without removing the filter. Concentration with Fluoropore FHLP and FGLP filters permitted the detection of single cells of microorganisms in 100-ml samples by PCR-gene probes.     

The effects of fungicide benomyl (benlate) on growth and mitosis in onion (Allium cepa L.) root apical meristem

In this study, the effects of benomyl, a systemic fungicide were investigated in the mitotic cell division in onion (Allium cepa) root tip cells during germination. For this aim, different concentrations (1, 2, 5, 10 and 20 mM) of benomyl solutions were used. All the concentrations used caused several abnormalities in mitotic cell divisions and the mitotic frequency in the onion root tip cells decreased as the concentration of benomyl solution increased. Based on our findings, it is reported that benomyl has some negative effects on mitotic divisions in onion root tip cells.