Comparative study of the level of mitotic activity and duration of mitosis over a 24 hour period in mouse tumors and normal tissue]

Circadian rhythm of cell division in the forestomack epithelium proved to be largely similar to that in the transplantable (continuous) carcinoma of the forestomack; the duration of mitosis in these tissues changed in the course of 24-hours. The mean 24-hour mitotic activity in the tumour was double that in the forestomack contrary to this, colchamine (colcemide) accumulated in the course of the 24-hours 121.1% mitoses in the forestomack and 83.8% mitoses in the carcinoma. A greater number of mitoses in the tumour with the usual count is attributed to the fact that the mean 24-hour duration of mitosis of the remen carcinoma was 2.7 greater than in the epithelium of the forestomack.     

Diurnal changes in the duration of mitosis in rats]

The colchicine method was applied to the study of the 24-hour changes in the duration of mitosis in 45-day rats. The average diurnal duration of mitosis in the pancreatic epithelium, the liver and the epidermis of adult animals was almost half of that in the 7-day-old rats. Diurnal variations of the mitotic index in the investigated tissues could be due both to the variation in the mitotic rate and to that of the duration of mitosis.     

Study of the effect of cAMP on the mitotic regimen in the esophageal epithelium of tumor-bearing mice]

The mitotic activity and number of DNA-synthesizing cells in the epithelium of the esophagus of the tumour-bearing albino mice were studied for 24 hours after the injection of dibutyryl cyclic 3', 5'-AMP. It was shown that injection of the preparation led to the blocking of cells in the G2-phase of the mitotic cycle, and to prolongation of mitosis during the first hours of the experiment without changing the total number of cells undergoing mitosis in the course of 24 hours.     

Duration of mitosis in mouse thymus cortex cells under normal conditions and after antigen administration at different times of the day]

The influence of an antigen injected at different periods of the circadian mitotic activity on the cell proliferation in the thymus cortex was studied. The duration of mitosis and the number of cells entering division were determined. A pronounced stimulating effect of immunization with sheep red blood cells entering mitosis increased, while the duration of mitosis diminished (colchamine mitotic index was 29.79 percent in control mice, and 47.99 percent in the immunized ones. The duration of mitosis was 0.4 hours in control animals, and 0.24 hours in the immunized ones. When compared with intact mice, the animals immunized in the evening showed no significant changes in the parameters studied.     

Participation of peptide hydra morphogen in the regulation of proliferation processes in different tissues of white rats]

The PMH influence on proliferation processes in some tissues of white rats was studied. PMH injections in doses 10.0 mkg/kg and 100.0 mkg/kg stimulated the processes of DNA--synthesis after 24 hours in corneal and tongue epithelium. The direct dose-dependent effect was revealed. PMH in dose 10.0 mkg/kg activated the proliferative processes in the stomach epithelium and thymus cortex. after 24 hours too. The given dose of PMH caused trustworthy decreasing of proliferative activity in thymus after 4 hours. The stimulation of DNA-synthesis and acceleration of mitosis were found after 5-times application on white rat's corneal with 100 nM PMH solution.     

The synchronizing effect of cyclic 3',5'-adenosine monophosphate on the mitotic cycle of Ehrlich ascitic carcinoma cells]

A study was made of the number of mitoses and of the DNA-synthesizing cells in the ascitic Ehrlich carcinoma in the course of 24 hours after the injection of cyclic 3',5'-adenosinmonophosphate to mice. It was found that as the result of the preprrophase inhibition and, possibly, of stimulation of the cell entry into the S-phase, 8 hours after the action a great number of cells began to divide almost simultaneously. The effect of mitosis synchronization was assessed in the tumour cell population.     

Circardian rhythm of mitoses in the epithelium of the cornea after splenectomy]

In corneal epithelium of CBA mice the index of colchicine mitoses diminished after splenectomy in the day period characterized by rising mitotic activity in control animals. The duration of active phase of cell division rhythm shortened while the maximum of mitotic activity delayed in comparison with control animals. The total amount of cells entering mitosis during 24 hours diminished by 27.7% and the rate of physiological regeneration of corneal epithelium decreased.     

Growth dynamics of an amphibian tissue

By the “labeled mitoses” method of Quastler and Sherman and others, the cell cycle of the germinative zone cells of the bullfrog lens epithelium has been characterized. It has been shown that this cycle lasts approximately 83 days with the DNA synthetic phase enduring 100 hours and G₂, 11 hours. G₁ occupies over 90% of the total time. the duration of mitosis itself has not been precisely determined. the length of the synthetic phase was corroborated by double labeling with c¹⁴ and h³-thymidine. When the temperature is raised by 6°c, from 24° to 30° the cycle is compressed by 40%. When the nongerminative, central cells of bullfrog lens epithelium are activated (stimulated to undergo DNA synthesis and mitosis) by injury or through in vitro culture, the length of the cycle also appears to decrease. in the in vitro experiments the generation time, as judged by the period elapsing between two successive bursts of DNA synthesis involving the same cells, amounts to 177–190 hours at 24°c. by raising the temperature to 30°c the time from injury or isolation until the appearance of the first wave of mitosis is reduced by 20%.     

The effect of 5-fluorouracil on the mitotic cycle and DNA synthesis in the epithelium of mouse small intestine.

The examinations were performed on 42 mice of the Porton strain. The experimental animals were injected intraperitoneally with the dose of 75 mg of 5-fluorouracil per kg body weight. The first experimental group received injections of [3H]thymidine within 48 hours and the second group within 96 hours of the injection of 5-fluorouracil. Two mice from each group were killed at within 1, 2, 4, 8, 12, 24, and 48 hours of the [3H]thymidine injection. Calculations of the mitotic index and time of duration of individual phases of the mitotic cycle in epithelial cells of the small intestine were based on application of the autoradiographic method. These studies lead to the conclusion that 5-fluorouracil disturbs the course of metabolic processes in the cell, which are also related with the distribution of the genetic material. Histological examinations show that 5-fluorouracil produces profound morphological changes in the intestine, which affect both the intestinal epithelium and the connective tissue stroma. The autoradiographic tests revealed a considerable suppression of the mitotic activity of the epithelium of intestinal crypts. Moreover, it was shown that 5-fluorouracil inhibits the mitotic activity of the intestinal epithelium by diminishing the number of cells capable of entering into mitosis. Nevertheless, by 96 hours following introduction of a single dose of 5-fluorouracil normal morphological structure and mitotic activity of the intestinal wall cells are restored.     

Inclusion body disease (herpesvirus infection) of falcons (IBDF).

Inclusion body disease of falcons (IBDF) is caused by a herpesvirus. The clinical course is short, 24 to 72 hours in duration, and is characterized by mild to severe depression and weakness often accompanied by anorexia. The disease is invariably fatal. The virus has a marked affinity for the reticuloendothelial system and hepatocytes,producing focal to diffuse necrosis of infected tissues accompanied by the formation of intranuclear inclusion bodies. The virus is pathogenic for American kestrels (Falco sparverius) and great horned owls (Bubo virginianus) in which typical lesions of IBDF are reproduced. The lesions of IBDF are similar to those produced by some herpesvirus infections in other avian species.     

Cell population kinetics and the cell population mechanisms of the circadian rhythm of proliferative activity in mouse esophageal epithelium]

The dynamics of 3H-thymidine labeled mitosis and diurnal rhythm of proliferative activity was studied. The isotope was injected to BALB/C mice at the peak of diurnal rhythm of DNA synthesis activity of basal layer cells of oesophageus epithelium. It has been established that the increase in the mitotic index during 24 hours depends on the increase in number of cells being in S-period. The data show that the increase of mitotic index at diurnal rhythm occurs at the expense of 75% of new G0-cells which entered into the mitotic cycle, and of 25% of re-entering cells that had divided during the maximal mitotic activity a day before. It is found that the duration of mitotic cycle of cell population which entered into the mitotic cycle synchronously is almost equal to the period of diurnal rhythm of mitotic activity, i.e. 24 hours.     

Effects of vinblastine, oncodazole, procarbazine, chlorambucil, and bleomycin in vivo on colchicine binding activity of tubulin.

Five chemotherapeutic agents which inhibit mitosis caused an $\text{in vivo}$ effect on the colchicine bound to the isolated tubulin from mouse lymphoma L5178Y cells. These effects were examined over a time course of 4, 12, 24, 48, and 72 hours after a single administration of each drug. Vinblastine, oncodazole, and bleomycin decreased the amount of colchicine bound per mg protein; procarbazine and chloroambucil increased the amount bound. All of the drugs except procarbazine required more than 4 hours to cause an effect on colchicine binding and in the case of bleomycin and oncodazole some recovery occurred after 48 hours. The mitotic index was affected by 4 hours by all drugs: procarbazine, chlorambucil and bleomycin caused a decrease; vinblastine and oncodazole, an increase.     

Effect of combined administration of estradiol and progesterone on the mitotic cycle of the epithelial tissues of the reproductive tract of ovariectomized white rats

The duration of stages of the cell cycle in the uterine and vaginal tissues of ovariectomized rats, treated with estradiol and estradiol-progesterone, was estimated using the labeled mitosis method. The joint treatment shortened G2 period in epithelial tissues. In the uterine epithelial tissues of estradiol-progesterone treated rats, the duration of S period was prolonged. In all tissues, progesterone stimulated the entry of cells into a second round of DNA synthesis. The estrogen-gestagen treatment inhibited the movement of vaginal epithelial cells from basal to superficial layers.     

Heterogeneity in G2 duration during lateral root development

Developing lateral roots of V. faba were treated with 5-aminouracil for up to 6 hours using the 5-AU inhibition method discussed in this paper; the duration of G₂+mitosis/2 and the percentages of slow dividing cells were estimated from the fall in MI observed in just emerged meristems, very large primordia and large primordia. The results indicate that during the period of development studied here there are two subpopulations of dividing cells: 1) fast dividing population which makes up about 84 % of the dividing cells and which has a G₂+mitosis/2 duration of about 3.3 hours, and 2) a slow dividing population which constitutes about 16 % of the dividing cells and which has a G₂ duration in excess of 12 hours. This heterogeneity is discussed in relationship to the behaviour of different populations of proliferating cells during root morphogenesis.     

Effect of thyroxine on the duration of the phases of the hepatocyte mitotic cycle at different times of day]

The lengths of the synthetic phase (S) and postsynthetic gap plus a half of the mitotic time (G2+1/2 M) has been investigated in hepatocytes of control and thyroxine-treated male white rats using percent labeled mitosis curves after injection of isotope at 10, 16, 22 and 4 o'clock. In the control, the minimum lengths of G2 lasted 3.0 hours without being changed during 24 hours. On the contrary, G2+1/2 M and S varied from 3.2 to 4.4 and from 8.0 to 9.5 hours, accordingly. A prolonged administration or hormone induced changes in duration of all the above phases whose alterations in thyroxine-treated group of animals showed 2.0--3.0, 2.9--3.4 and 6.4--11.3 hours, respectively. During 24 hours, there was observed a characteristic pattern of changes in the labeling index (LI) of both groups of animals. It has been established for both the groups that the increased in LI coincides with the shortening of S-phase. The data allow to conclude that some intracycle mechanisms may exist controlling the cell division and exerting their effects on the cells at the end of G1-phase and during G2-phase. Thyroxine is a regulator of cell proliferation, and its effect was found to occur due to the intracycle mechanisms of cell cycle kinetics.     

DNA synthesis and mitosis in a temperature sensitive Chinese hamster cell line

Viability, DNA synthesis and mitosis have been followed in the temperature sensitive Chinese hamster cell mutant K12 under permissive and non-permissive conditions. On incubation at 40°C cells retained their ability to form colonies at 33°C for 15 to 20 hours, but viability was lost gradually during the following 20 hours. When random cultures of K12 were shifted to 40°C the rate of DNA synthesis was normal for three to four hours but then decreased markedly, reaching 95% inhibition after 24 hours. Under the same conditions mitosis was inhibited after 15 hours. If cultures which had been incubated at 40°C for 16 hours were placed at 33°C the rate of DNA synthesis increased five hours after the shift down and mitosis 18 hours after. These results can be interpreted on the assumption that K12 at 40°C is unable to complete a step in the cell cycle which is essential for DNA synthesis and which occurs three to four hours before the start of S at 33°C.     

Regulation of a COPII component by cytosolic O-glycosylation during mitosis

Endoplasmic reticulum (ER)-to-Golgi transport is blocked in mammalian cells during mitosis; however, the mechanism underlying this blockade remains unknown. Since COPII proteins are involved in this transport pathway, we investigated at the biochemical level post-translational modifications of COPII components during the course of mitosis that could be linked to inhibition of ER-to-Golgi transport. By comparing biochemical properties of cytosolic COPII components during interphase and mitosis, we found that Sec24p isoforms underwent post-translational modifications resulting in an increase in their apparent molecular weight. No such modification was observed for the other COPII components Sec23p, Sec13p, Sec31p or Sar1p. Analyzing in more details Sec24p isoforms in interphase and mitotic conditions, we found that the interphase form of Sec24p was O-N-acetylglucosamine modified, a feature lost upon entering into mitosis. This mitotic deglycosylation was coupled to Sec24p phosphorylation, a feature likely responsible for the increase in apparent molecular weight of these molecules. These modifications correlated with an alteration in the membrane binding properties of Sec24p. These data suggest that when entering into mitosis, the COPII component Sec24p is simultaneously deglycosylated and phosphorylated, a process which may contribute to the observed mitotic ER-to-Golgi traffic block.     

Nutritional Requirements for Polyploid Mitoses in Cultured Pea Root Segments

Diploid and polyploid mitoses could be stimulated in excised segments of the mature region of pea roots grown on a sterile culture medium. Diploid mitoses were observed in segments cultured on water alone for 72 hours. Their frequency was increased by the presence of salts, sucrose, vitamins, and any two or all three of the following: an amino mixture, auxins, and kinetin. Polyploid mitoses were observed 72 hours after the beginning of the culture period in segments cultured on salts, sucrose, vitamins, auxins, and kinetin. Polyploid mitoses required the presence of auxins and kinetin in the culture medium. Their frequency was not affected by the presence of a reduced nitrogen source. Light treatments had no effect on the frequency of diploid or polyploid mitoses. Diploid mitoses were first observed about 24 hours after the beginning of the culture and their frequency increased thereafter. Experiments with colchicine showed that diploid cells were entering mitosis for the first time as late as 60 hours after the beginning of the culture. Polyploid mitoses showed a long lag time when compared with diploid mitoses. They began at about 60 hours and their frequency increased thereafter. Experiments with colchicine showed that polyploid cells were entering mitosis for the first time as late as 84 hours after the beginning of the culture. The presence of kinetin in the medium was not required during the first 24 hours in culture for the appearance of polyploid mitoses at 74 hours. However, the presence of kinetin was required after 24 hours. Auxin was required at some time during the first 24 hours of the culture and its continuous presence may be required for the stimulation of polyploid mitoses.     

Effects of serum deprivation on the initiation of DNA synthesis in the second generation in rat 3Y1 cells

Rat 3Y1 cells arrested at early S by hydroxyurea traversed the remainder of S and G2 and completed mitosis after removal of the drug, irrespective of the absence of serum from the culture medium. When cells were deprived of serum for a period between early S and mitosis after removal of hydroxyurea, the cells delayed entry into S in the presence of serium in the second generation for the time length approximately equal to that of serum deprivation. When mitotic cells, which had been continously exposed to serum after removal of hydroxyurea, were deprived of serum for the next 24 hours and then were reexposed to serum, the cells delayed entry into S for more than 24 hours (more than the time length of serum deprivation). On the other hand, the cells already deprived of serum between early S and G2 in the first generation were less delayed in entry into S after postmitotic 24-hour serum deprivation than were the cells exposed to serum between early S and G2 in the first generation. These results suggest that serum-dependent events continue to occur in the first generation for on-time entry into S in the next generation, and that these premitotic events (the potential for entry into S) decay if serum is absent for a long period of time after mitosis.     

MITOTIC ACTIVITY AT THE SHOOT APEX OF AZOLLA FILICULOIDES

The mosquito fern, Azolla filiculoides Lam., was grown in a growth chamber on a nitrogen-free culture solution at 24 C under the following photoperiod: 16 hr light/8 hr darkness. Shoot tips were fixed every 2 hr for 24 hr to determine the mitotic index for the apical cell, immediate derivatives, and remaining cells to the level of the first leaf or lateral shoot primordium. Mitotic indices were 6.9%, 6.5% and 6.3%, respectively. The colchicine method was employed to determine the cell-cycle durations and duration of mitosis for the same populations of cells. The cell-cycle duration and duration of mitosis of the apical cell were 28.2 hr and 2.8 hr, respectively; for the immediate derivatives, 26.7 hr and 2.5 hr; for the remaining cells, 23.6 hr and 2.1 hr. Conclusions: the apical cell is as mitotically active as its immediate derivatives, and there is no evidence of a quiescent apical cell.