Time-Lapse Cinemicrographic Studies of X-Irradiated HeLa S3 Cells: I. Cell Progression and Cell Disintegration

Time-lapse cinemicrographs of synchronous HeLa S3 cells irradiated with 220 kv X-rays at various stages of interphase provided data for constructing pedigrees, measuring the duration of both generation cycles and mitoses, and scoring events associated with cell disintegration for up to seven postirradiation generations. The onset of the first mitosis after doses of 500 rads was delayed as expected from previous studies of the age dependence of “mitotic delay.” The interval between this first mitosis and the next was indistinguishable from that for unirradiated control cells, while the subsequent two generations were again prolonged, on the average, though not so severely as was the irradiated generation. The duration of mitosis was increased proportionally more than interphase. Cell disintegration took place by way of two morphologically distinct processes. In three-quarters of the cases the cells were rounded and apparently trapped in metaphase when they disintegrated; the remaining disintegrations occurred in spread, interphase cells. In cells disintegrating from the rounded configuration, the generation preceding disintegration was prolonged relative to that in cells which divided; in cells disintegrating from either configuration, the penultimate generation was also prolonged. The mitotic times were disproportionately increased in both of these generations. It is suggested that in this system X-ray damage is preferentially expressed as derangement of the mitotic process; such damage ultimately brings about permanent mitotic arrest in the majority of cells.     

Fluctuation periodicity,generation separation,and the expression of larval competition

A suite of models has been formulated to investigate the dynamic consequences of the various routes by which uniform larval competition for food can find demographic expression. It is found that while delayed expression through the vital rates of later age classes gives rise to limit cycles containing multiple overlapping generations, immediate expression via changes in the death or growth rates of the larvae themselves leads to self-sustaining single generation limit cycles. When immediate expression of competition is combined with high adult fecundity and short reproductive lifespan the amplitude of the single generation cycles is so large that they constitute a series of evenly spaced discrete generations, which is maintained indefinitely even in the absence of external cues.     

Renewal of the epithelium in the descending colon of the mouse. IV. Cell population kinetics of vacuolated-columnar and mucous cells.

Proliferation and migration of cells in the vacuolated-columnar and mucous cell lines were studied in the descending colon of adult female mice given a single injection or a continuous infusion of 3H-thymidine and killed at various intervals from one hour to 12 days. This investigation was carried out using one mum-thick Epon sections which were radioautographed after staining with the periodic acid-Schiff technique and iron-hematoxylin. In the normalized crypts with ten equal segments, labeled vacuolated cells at one hour after injection of 3H-thymidine were encountered in the lower four segments and in decreasing numbers in segments 5 through 7. From the percent labeled cells in segments of the crypt, the birth rate and fluxes of cells were computed. Moreover, it was found that a cell in the vacuolated-columnar cell line would undergo three mitotic cycles on the average from its birth at the cryptal base to its extrusion from the surface; of these three cycles, the last one which took place from segment 3 to segment 7 appeared to be a changeover from dividing cells to non-dividing cells, in accordance with the "slow cut-off" model of Cairnie et al. ('65b). Mucous cells located in segments 1 through 6 of the crypt were capable of incorporating 3H-thymidine and thus capable of undergoing mitosis. However, the rate of turnover of mucous cells based on proliferative rate was found to be much lower than the rate of turnover of mucous cells based on the transit time in the non-dividing segments of the crypt. Since there was a concomitant overproduction of cells in the vacuolated cells and newly formed mucous cells in the lower portion of the crypt, it was concluded that some vacuolated cells would give rise to mucous cells. This putative transformation occurred in the lower four segments of the crypt. Mucous cells which were formed by transformation would migrate upward along the cryptal wall and accumulate more mucus in the theca; in doing so, they would undergo two divisions, on the average, before they became non-dividing mucous cells. In ascending the cryptal walls, both vacuolated-columnar cells and mucous cells appeared to migrate at a similar speed; they moved much slower at the base of the crypt and accelerated toward the upper portion of the crypt, but they migrated at a constant speed in the non-dividing segments of the crypt.     

An analysis of the growth of the retinal cell population in embryonic chicks yielding proliferative ratios, numbers of proliferative and non-proliferative cells and cell-cycle times for successive generations of cell cycles

Growth curves of the retinal cell population of embryonic chicks were fitted by a branching-process model of cell population growth, thereby estimating the proliferative ratios and mean cell-cycle times of the generations of cell cycles that underlie retinal growth. The proliferative ratio determines the proportion of cells that divides in the next generation, so the numbers of proliferative and non-proliferative cells in each generation of cell cycles were obtained. The mean cell-cycle times determine the times over which the generations are extant. Assuming growth starts from one cell in generation 0, the proliferative cells reach 3.6 × 10⁶ and the non-proliferative cells reach 1.1 × 10⁶ by generation 23. The next four generations increase the proliferative cell numbers to 13.9 × 10⁶ and produce 20.1 × 10⁶ non-proliferative cells. In the next five generations in the end phase of growth, non-proliferative cells are produced in large numbers at an average of 13.9 × 10⁶ cells per generation as the retinal lineages are completed. The retinal cell population reaches a maximum estimated here at 98.2 × 10⁶ cells. The mean cell-cycle time estimates range between 6.8 and 10.1 h in generations before the end phase of growth and between 10.6 and 17.2 h in generations in the end phase. The retinal cell population growth is limited by the depletion of the proliferative cell population that the production of non-proliferative cells entails. The proliferative ratios and the cell-cycle-time distribution parameters are the likely determinants of retinal growth rates. The results are discussed in relation to other results of spatial and temporal patterns of the cessation of cell cycling in the embryonic chick retina.     

PROLIFERATION KINETICS IN EPITHELIUM OF GUINEA-PIG COLON III. DISTRIBUTION OF BLOOD CAPILLARIES ALONG THE CRYPT

The colonic blood vessels of the guinea-pig were infused with carmine-gelatine and the occurrence along the crypt of mucosal capillaries containing the dye was examined. The pattern of capillary distribution along the crypt is inhomogeneous but is similar for short, medium and long crypts. Capillaries occur least frequently at the bottoms of crypts, that is within the lowest 0.05% of the crypt length. The frequency rises significantly between 0.05 and 0.40 of the crypt length; this rise is accompanied by an increase in the number of DNA-synthesizing cells. A significant fall in capillary frequency along the crypt between 0.35 and 0.45 of crypt length coincides with the beginning of a decrease in the number of DNA-synthesizing cells. The highest frequency of capillaries occurs in the upper 0.20 of the crypt length and beneath the lining epithelium.     

大猿叶虫龙南种群的生物学特性

系统研究大猿叶虫Colaphellus bowringi Baly江西龙南(24°9′N,114°8′E)种群生物学特性。结果显示,该虫仅在春季和秋季发生为害,以成虫在土中越冬和越夏。由于成虫滞育期的差异,该虫显示出生活史多样性,有些个体隔年繁殖;有些个体是一化性,仅在春季或秋季繁殖1代;有些个体是二化性,在春季和秋季各繁殖1代;有些个体是多化性的,春季1代,秋季2~3代,因此,在田间1年可发生1~4代。春季,滞育成虫于2月中旬至4月初陆续出土繁殖,羽化的成虫于4月上旬至5月中旬陆续入土越夏;秋季,滞育成虫于8月中旬至10月中旬陆续出土,羽化的成虫于9月中旬至12月中旬陆续入土越冬。成虫一生能多次交配,在25℃下,春季世代雌虫产卵期为8~29d,平均产卵量为776粒;秋季世代雌虫产卵期为9~39d,平均产卵量为1003粒。各虫态的发育历期:在18~28℃间,卵为8.66~3.85d,幼虫为19.37~7.92d,蛹为8.57~3.41d。卵、幼虫和蛹的发育起点温度分别为10.5、11.5和11.9℃。滞育成虫的寿命为5~28个月。     

Observations on the endogenous stages of Eimeria crandallis in domestic lambs (Ovis aries)

Lambs reared coccidia-free were inoculated orally with various numbers of sporulated oocysts of E. crandallis and were killed between 1 and 22 days after inoculation; tissues were examined histologically. Sporozoites were seen 1, 2 and 3 days after inoculation (DAI) in crypt epithelial cells in the mid-jejunum. Infected cells migrated into the lamina propria where the parasite within them developed into a firstgeneration meront containing about 250,000 merozoites at 10 DAI. A second generation of meronts was seen at 10–12 DAI, each containing up to about 10 merozoites, situated mainly at the bases of crypts in the jejunum and ileum but also in the caecum. From 11 DAI pro-gamonts were seen which were enveloped by the host cell nucleus and which divided in synchrony with the host cell for an undetermined number of generations. Mature gamonts began to develop from them by 16 DAI. Oocyst output began at 16 DAI and rose to a peak at about 22 DAI. Up to 10⁸ oocysts were produced per oocyst inoculated. They showed wide variation in size and colour.     

Observations on the endogenous stages of Eimeria crandallis in domestic lambs (Ovis aries)

Lambs reared coccidia-free were inoculated orally with various numbers of sporulated oocysts of E. crandallis and were killed between 1 and 22 days after inoculation; tissues were examined histologically. Sporozoites were seen 1, 2 and 3 days after inoculation (DAI) in crypt epithelial cells in the mid-jejunum. Infected cells migrated into the lamina propria where the parasite within them developed into a firstgeneration meront containing about 250,000 merozoites at 10 DAI. A second generation of meronts was seen at 10–12 DAI, each containing up to about 10 merozoites, situated mainly at the bases of crypts in the jejunum and ileum but also in the caecum. From 11 DAI pro-gamonts were seen which were enveloped by the host cell nucleus and which divided in synchrony with the host cell for an undetermined number of generations. Mature gamonts began to develop from them by 16 DAI. Oocyst output began at 16 DAI and rose to a peak at about 22 DAI. Up to 10⁸ oocysts were produced per oocyst inoculated. They showed wide variation in size and colour.     

Analysis of intestinal cell proliferation after guanethidine-induced sympathectomy. II. Percentage labelled mitoses studies

Neonatal administration of guanethidine-sulfate results in an alteration of the cell proliferative pattern of the small intestinal epithelium of the young adult rat. Sympathectomy with guanethidine has previously been shown to depress mitotic, labelling, and total cellular migration indices while increasing the generation cycle time (Tc) of small intestinal crypt cells as measured by a stathmokinetic method. The present study showed that the G1, S and G2 phases of the crypt cell cycle are altered by sympathectomy, G1 accounting for most of the increase in Tc. In addition, the percentage of [3H]-thymidine labelled crypt cells is reduced and the duration of crypt cell transit is lengthened by guanethidine-induced sympathectomy.     

Early evolution of life cycles in embryophytes: A focus on the fossil evidence of gametophyte/sporophyte size and morphological complexity

Abstract Embryophytes (land plants) are distinguished from their green algal ancestors by diplobiontic life cycles, that is, alternation of multicellular gametophytic and sporophytic generations. The bryophyte sporophyte is small and matrotrophic on the dominant gametophyte; extant vascular plants have an independent, dominant sporophyte and a reduced gametophyte. The elaboration of the diplobiontic life cycle in embryophytes has been thoroughly discussed within the context of the Antithetic and the Homologous Theories. The Antithetic Theory proposes a green algal ancestor with a gametophyte‐dominant haplobiontic life cycle. The Homologous Theory suggests a green algal ancestor with alternation of isomorphic generations. The shifts that led from haplobiontic to diplobiontic life cycles and from gametophytic to sporophytic dominance are most probably related with terrestrial habitats. Cladistic studies strongly support the Antithetic Theory in repeatedly identifying charophycean green algae as the closest relatives of land plants. In recent years, exceptionally well‐preserved axial gametophytes have been described from the Rhynie chert (Lower Devonian, 410 Ma), and the complete life cycle of several Rhynie chert plants has been reconstructed. All show an alternation of more or less isomorphic generations, which is currently accepted as the plesiomorphic condition among all early polysporangiophytes, including basal tracheophytes. Here we review the existing evidence for early embryophyte gametophytes. We also discuss some recently discovered plants preserved as compression fossils and interpreted as gametophytes. All the fossil evidence supports the Antithetic Theory and indicates that the gametophytic generation/sporophytic generation size and complexity ratios show a gradual decrease along the land plant phylogenetic tree.     

The influence of various cell kinetic conditions on functional differentiation in the small intestine of the rat. A study of enzymes bound to subcellular organelles

The process of cell maturation and cell ageing of absorptive epithelial cells was investigated in normal rat duodenum. The development of a number of enzymes bound to subcellular organelles was studied by using microchemical analyses on various cell compartments dissected from crypts and villi from freeze-dried cryostat sections. The development of the ultrastructural features of the absorptive epithelium was investigated by electron microscopy of various cell positions along the whole length of the crypt and the base of the villus. The data obtained were related to cell position along the crypt and villus and to cell age during migration from the bottom of the crypt to the tip of the villus.The influence of changes in the life-span of the cells and of increasing proliferative activity was studied by comparing normal rat duodenum with that from germfree rats and rats recovering from low radiation doses (72 hr after 400 R).Our data show that the specific activity of nonspecific esterases mainly localized in the endoplasmic reticulum increases when the cells migrate along the upper half of the crypt and the basal part of the villus. Activity of alkaline phosphatase, measured as a marker for the microvilli, is absent in the crypt, but increases linearly from the base of the villus to the tip. The longer life-span of villus cells in germfree animals does not result in a higher activity of these enzymes than in normal animals. An increased proliferative activity in the crypt, as present 72 hr after X-irradiation, is accompanied by a decreased activity of both enzymes but the pattern of activity during cell migration remains the same. The specific activity of enzymes bound to mitochondria or lysosomes (monoamineoxidase and β-N-acetylglucosaminidase) are not affected by changing crypt cell kinetics.Electrophoretic analyses of isolated cell compartments showed that the increase during normal differentiation or the decrease after X-irradiation of esterase activity is due to changes in overall activity, not to the appearance or disappearance of specific isoenzymes. Electron microscopy showed that in the normal intestine there is a gradual development of ultrastructural features during migration of the cell along the crypt while the most drastic changes in cell structure occur at the moment the cell enters the villus. Contrary to our expectation, the ultrastructural development was not influenced by increased proliferative activity in the crypt 72 hr after irradiation, and hence the decrease in enzyme activity found cannot be related to changes in ultrastructure.     

不同小麦品种对有翅与无翅型禾谷缢管蚜生长发育与繁殖的影响

室内采用水培小麦法饲养禾谷缢管蚜Rhopolosiphum padi(L.),比较了有翅与无翅型禾谷缢管蚜若虫发育历期、成虫寿命、繁殖力和日均体重增长量.结果表明,不同小麦品种上有翅型和无翅型若虫发育历期均存在显著差异;同时产若蚜数量、每代产若蚜数和腹中胚胎数均存在显著差异,而成蚜寿命、产若蚜历期和产若蚜代数差异不显著...     

上海地区悬铃木方翅网蝽的生活史及发生情况

观察悬铃木方翅网蝽Corythucha ciliata(Say)在上海的生活史和生活习性,并对其在上海的发生情况进行调查。悬铃木方翅网蝽在上海1年发生5代,第5代成虫于10月中下旬主要在悬铃木主干和支干的树皮下越冬,4月上旬越冬成虫上树为害,4月下旬开始产卵,5月上旬始见第1代若虫,第1代若虫期17～18d,世代历期约30d,7～8月间世代重叠现象明显。悬铃木方翅网蝽在上海11个区均有发生,株被害率最高的4条道路分别为徐汇区龙华西路、肇嘉浜路、松江区人民北路及长宁区延安西路,为害率达100%。为害程度最严重的的3条道路分别为松江区人民北路、长宁区延安西路和水城南路,虫情指数分别达95.6%、76.7%和72.2%。     

Eimeria vermiformis: host strains and the developmental cycle

After confirmatory studies of the endogenous development of Eimeria vermiformis in hybrid mice, a comparison, both qualitative and quantitative, was made of the developmental cycle of the parasite in two strains of host of contrasting susceptibility. No differences were apparent between the resistant, BALB/c, and susceptible, C57BL/6, during the first 5 days of the cycle. Thereafter, more parasites were seen in the C57BL/6, they were more widely distributed along the intestine, and they persisted in the tissues for a considerably greater time. Schizogony continued for longer in the C57BL/6 mice, concurrently with gametogony, and there were probably up to four additional generations of schizonts, which resembled those of the accepted third generation. The kinetics of oocyst production, measured in the feces, reflected events in the intestinal epithelium. The findings are discussed in relation to eimerian life cycles and the immune response.     

Dynamics in Central European near‐natural Abies‐Fagus forests: Does the mosaic‐cycle approach provide an appropriate model?

Question: The mosaic‐cycle concept of forest dynamics dominates in Central Europe. According to this concept intermediate‐scale disturbances only accelerate the forest break‐up under existing cycles of forest development. Is such an approach correct, or should new developmental cycles be elaborated for intermediate‐scale disturbances? Location: Near‐natural Abies alba ‐ Fagus sylvatica forests in the ?wiétokrzyski National Park in Central Poland. In these forests intermediate‐scale disturbances occurred between 1970 and 1990. Methods: Data were collected twice in areas surrounding 212 permanent sample points (in 1994 and 2004). Two increment cores were taken from 259 sample Abies trees. The effect of intermediate‐scale disturbances on radial increment of Abies was assessed. Probabilities of stand transition during a 10‐year period between individual stages and phases of development of the mixed forest were calculated. The development stages and phases were arranged into hypothetical succession series of successive changes. Results: In 1994 70 stands and in 2004, 47 stands representing stages and phases containing the older generation formed by trees > 100–150 years were found. Also, in 1994 142 and in 2004, 165 stands representing stages and phases containing the younger generations only, formed by trees < 100‐150 years, were recorded. Stages and phases containing only younger generations do not occur in the existing forest development cycle which does not consider the influence of intermediate‐scale disturbances separately. Two developmental cycles, which take into account the presence of the older generation and the younger generations only (under conditions of the occurrence of intermediate‐scale disturbances), are proposed. Conclusion: The mosaic‐cycle concept of forest dynamics can be used to analyse the dynamics of Central European near‐natural mixed‐species forests, but new developmental cycles should be elaborated for intermediate‐scale disturbances.     

Cholecystokinin- and dexfenfluramine-induced anorexia compared using devazepide and c-fos expression in the rat brain

It has been proposed that there might be a link between the anorectic actions of cholecystokinin (CCK) and serotonin (5HT). The present study compared the patterns of c-fos protein-like immunoreactivity (FLI) induced in rat brain by CCK and the indirect 5HT agonist dexfenfluramine (DFEN), as well as the ability for devazepide, a CCK-A receptor antagonist, to antagonize both anorexia and FLI induced by these agents. Devazepide reversed the anorectic effect of CCK but not that of DFEN in food deprived rats. The FLI induced by CCK and DFEN occurred in similar brain regions, but in different subdivisions. Such regions included the bed nucleus of the stria terminalis (BST), the lateral central nucleus of the amygdala (CeL), and the lateral parabrachial nucleus (LPB). Devazepide abolished the FLI induced by CCK most of these brain regions, but had no effect on FLI induced by DFEN. These results suggest that the LPB-CeL/BST pathway might be responsible for the anorectic effects of both CCK and DFEN, but different parts or neuronal populations in these structures might be differentially engaged by CCK and DFEN. The putative interaction between CCK and 5HT might happen along this pathway, rather than in the periphery.     

Fluctuating thermal environments and time‐dependent effects on fruit fly egg‐hatching performance

Organismal performance in a changing environment is dependent on temporal patterns and duration of exposure to thermal variability. We experimentally assessed the time‐dependent effects of thermal variability (i.e., patterns of thermal exposure) on the hatching performance of Drosophila melanogaster. Flies were collected in central Chile and maintained for four generations in laboratory conditions. Fourth generation eggs were acclimated to different thermal fluctuation cycles until hatching occurred. Our results show that the frequency of extreme thermal events has a significant effect on hatching success. Eggs exposed to 24 hr cycles of thermal fluctuation had a higher proportion of eggs that hatched than those acclimated to shorter (6 and 12 hr) and longer cycles (48 hr). Furthermore, eggs subjected to frequent thermal fluctuations hatched earlier than those acclimated to less frequent thermal fluctuations. Overall, we show that, egg‐to‐adult viability is dependent on the pattern of thermal fluctuations experienced during ontogeny; thus, the pattern of thermal fluctuation experienced by flies has a significant and until now unappreciated impact on fitness.     

Dual-wavelength imaging of tumor progression by activatable and targeting near-infrared fluorescent probes in a bioluminescent breast cancer model

Bioluminescence imaging (BLI) has shown its appeal as a sensitive technique for in vivo whole body optical imaging. However, the development of injectable tumor-specific near-infrared fluorescent (NIRF) probes makes fluorescence imaging (FLI) a promising alternative to BLI in situations where BLI cannot be used or is unwanted (e.g., spontaneous transgenic tumor models, or syngeneic mice to study immune effects).In this study, we addressed the questions whether it is possible to detect tumor progression using FLI with appropriate sensitivity and how FLI correlates with BLI measurements. In addition, we explored the possibility to simultaneously detect multiple tumor characteristics by dual-wavelength FLI (~700 and ~800 nm) in combination with spectral unmixing. Using a luciferase-expressing 4T1-luc2 mouse breast cancer model and combinations of activatable and targeting NIRF probes, we showed that the activatable NIRF probes (ProSense680 and MMPSense680) and the targeting NIRF probes (IRDye 800CW 2-DG and IRDye 800CW EGF) were either activated by or bound to 4T1-luc2 cells. In vivo, we implanted 4T1-luc2 cells orthotopically in nude mice and were able to follow tumor progression longitudinally both by BLI and dual-wavelength FLI. We were able to reveal different probe signals within the tumor, which co-localized with immuno-staining. Moreover, we observed a linear correlation between the internal BLI signals and the FLI signals obtained from the NIRF probes. Finally, we could detect pulmonary metastases both by BLI and FLI and confirmed their presence histologically.Taken together, these data suggest that dual-wavelength FLI is a feasible approach to simultaneously detect different features of one tumor and to follow tumor progression with appropriate specificity and sensitivity. This study may open up new perspectives for the detection of tumors and metastases in various experimental models and could also have clinical applications, such as image-guided surgery.     

Impact of dietary sterols on life-history traits of a caterpillar

Although lepidopteran larvae have been shown to metabolise a number of different phytosterols, their life-history traits in response to these different phytosterols may vary with the species. Using the specialist moth, Plutella xylostella (Linaeus) (Lepidoptera), growth, development and fecundity on artificial diets containing host and non-host phytosterols were measured over two generations. Because of the nature of the experimental design, maternal effects linked to dietary sterol could be assessed. In both generations, survival was highest on diets containing sitosterol as the primary phytosterol. A significant decrease in survival on stigmasterol, a non-host phytosterol, was observed between generations. As expected, survival on a control diet with only trace-sterols decreased between generations. Larval duration and pupal duration in both generations was longest on the diets containing the non-host phytosterols, stigmasterol and spinasterol, and on the trace-sterol diet. Finally, fecundity was affected by dietary sterol. In the first generation, fewer viable eggs were produced on the stigmasterol and trace-sterol diets than on wheatgerm control diet. In both generations, dietary sterol significantly influenced percentage egg viability. Overall, these results indicate that differences in phytosterol structure can have pronounced effects on life-history traits, especially when they are projected to the population level. The implications of these results are discussed with respect to phytosterol use by phytophagous insects and their host plant affiliations.     

Discrimination of different generations of Zeugodacus scutellata using age grading technique and their local genetic variation

The striped fruit fly, Zeugodacus scutellata, exhibits generation overlap due to its relatively long lifespan. This study clarified the fly generations by age-grading technique and traced migration of local populations using genetic markers. Male adults were monitored using Cue-lure in two different places (Andong and Guri) of mainland Korea during the entire host cultivating period. They showed two occurrence peaks (from late July to early August and from mid-August to mid-October). A developmental model of Z. scutellata using rate summation method predicted four generations during this period. Pterin amounts in fly heads increased with adult age and indicated at least 3 cycles of age variation during the occurrence period with an additional adult generation after the third cycle. The additional peak was supported by field monitoring using protein diet supplemented with terpinyl acetate, which showed an additional occurrence peak during October. These seasonal populations were monitored through their migration by gene flow analysis using randomly amplified polymorphic DNA (RAPD) markers. In each locality, three seasonal populations at July, August, and September were discrete. Although local populations were genetically discrete at July, they were not separate at late seasons except for Jeju populations. These results indicate the usefulness of the head pterin quantification in age-grading of adult populations and suggest that Z. scutellata occurs at least four generations a year. In addition, gene flow analysis using RAPD suggests frequent movements among local populations along with season progression.