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.     

A study of diurnal proliferative activity in tumour and small intestine of C3H mice bearing a transplanted mammary carcinoma

Diurnal changes in proliferative activity were investigated in tumour and small intestinal epithelium of mice bearing a transplanted mammary carcinoma. In addition to mitotic and labelling index studies, the metaphase-arrest technique with vincristine (VCR) was employed. In the tumour there was no clear evidence of a significant diurnal rhythm in proliferative activity but in the small intestinal epithelium such a rhythm was clearly demonstrated. A higher cell production rate (kB) measured by metaphase-arrest and higher labelling and mitotic indices were seen in the mid to late part of the dark period. The peak mitotic index was seen 3 to 6 h after the labelling peak in the small intestine. The basal third of the crypt which is believed to include the stem cell compartment of this tissue showed larger diurnal fluctuations in both labelling index and kB than the rest of the proliferative compartment.     

THE RELATIONSHIP BETWEEN DIURNAL VARIATION OF THE NUMBER OF CELLS IN MITOSIS AND OF THE NUMBER OF CELLS SYNTHESIZING DNA IN THE EPITHELIUM OF THE HAMSTER CHEEK POUCH

1. The numbers of cells in mitosis and in DNA synthesis in the epithelium of the hamster cheek pouch have been studied at different times of the day and night. 2. By accumulation of mitotic cells using colcemid, both the rate of entry of cells into mitosis and the duration of mitosis have been estimated at two different times of day. 3. A diurnal variation has been demonstrated in both the mitotic index and in the tritiated thymidine labelling index. Although these variations are of different amplitude and timing, the experimental data fit closely to the hypothesis that the diurnal mitotic variation is the result of a partially synchronous population moving through the DNA synthetic period. No direct action on the mitotic process need be postulated. 4. From the results of mitotic accumulation, it is clear that the rate of entry of cells into mitosis depends on the time of day at which this is studied. There is also the possibility that the duration of mitosis is slightly longer when the mitotic index is high. 5. It is concluded that, at least in the epithelium of the hamster cheek pouch, the diurnal rhythm in the number of mitoses present is a reflection of the diurnal variation in the number of cells synthesizing DNA at some time earlier. Small fluctuations in the mitotic pattern imposed by this partially synchronous population moving from S into mitosis, could be caused by slight variations in the duration of mitosis.     

Circadian rhythm of the mitotic activity and of the number of nuclei synthesizing DNA in sarcoma 37 in white mice]

Circadian variations in the frequency of mitoses and the number of nuclei labed with thymidine-H3 in sarcoma-37 of mice were investigated. It was shown that the circadian rhythm of mitotic activity was composed of diurnal variations in the frequency of labeled and unlabeled mitoses. The G2-phase of mitotic cycle of the cells with labeled mitosis was approximately one hour. The G2-phase of the cells with unlabed mitosis lasted four hours and more. It is suggested that there are two cell populations in sarcoma-37.     

DIURNAL VARIATION IN THE LABELING INDEX OF MOUSE EPIDERMIS: A DOUBLE ISOTOPE AUTORADIOGRAPHIC DEMONSTRATION OF CHANGING FLOW RATES

A diurnal rhythmicity in the labeling index was observed in the epidermis of hairless mice, injected with either ¹⁴C- or ³H-thymidine, at different times during a 24 hr period. A modified autoradiographic technique, using ¹⁴C- and ³H-thymidine and two overlying emulsion layers, makes it possible to clearly differentiate synthesizing cells which are singly labeled with either carbon-14 or tritium, and cells labeled with both isotopes. At various times during a 24 hr period, hairless mice were injected with thymidine-2-¹⁴C and colcemid, followed at 2 or 3 hr by a second injection of ³H-thymidine. The labeling indices were calculated for the ¹⁴C- and ³H-thymidine injection times. These labeling indices were consistent with the control, single isotope, labeling indices and exhibited the same diurnal rhythm. Cells singly labeled with ³H- or ¹⁴C-thymidine have either started or completed DNA synthesis during the interval between the two injections. Flow rates into and out of DNA synthesis, throughout the 24 hr period, can be calculated from these singly labeled cells. The flow rates varied rhythmically throughout the day and paralleled changes in the labeling indices. The influx and efflux flow rates, at all times measured, were not equal. The influx flow rate was reflected in the efflux rate at a time later equal to the duration of S. By means of these flow rates, the per cent of cells in DNA synthesis was calculated for each hour during a 24 hr period. The resulting labeling index curve matches the observed 24 hr diurnal rhythm in labeling indices. By extension of these flow rates through mitosis, the resulting mitotic index curve is comparable to the reported 24 hr diurnal rhythm in mitotic indices.     

Effect of thyroxine on the diurnal rhythm of the mitotic activity and parameters of the life cycle of cells of the liver and esophagus]

The duration of the cellular cycle and the diurnal rhythm of the amount of mitosis were studied in young rats in normality and under the influence of thyroxin. The parenchymal and connective-tissue cells of the liver and cells of the liver and the cells of the oesophagus epithelium basal layer were studied. It was found that under the influence of thyroxin there occured a shortening of the periods of the cellular cycle and a 3--6 h shift to the left of the diurnal rhythm curve of the amount of mitoses. In thyroxinized animals the 21--95% increase of the amount of mitoses in the period of maximum values of the mitotic index during a day was observed as compared with control animals. A conclusion is made about the diurnal rhythm of sensitivity of G0-phase cells to the synchronizing factor, suggesting the decisive significance of the state of the cell population in the interaction of the tissue and hormone cells. The data obatained in the work show that the thyroid hormones regulate the cellular reproduction in the organism by stimulating the cells in the division cycle, synchronization of greater amount of cells by the moment of beginning of the mitotic cycle at a definite time of day and by shortening the period of the cell mitotic cycle.     

MITOTIC ACTIVITY IN THE ROOT APEX OF THE WATER FERN MARSILEA VESTITA HOOK. AND GREV.

Roots of Marsilea vestita ranging from 1–120 mm in length, as well as root primordia, were analyzed to determine mitotic activity and ploidy levels in the apical cell, five well-defined regions of the root proper, and two regions in the root cap. The mitotic index of the apical cell tended to be above the overall mean mitotic index for the entire apical meristem. No diurnal rhythm in mitotic index was apparent. The cell-cycle duration of the apical cell ranged from 12.1–25.2 hr, that of other regions of the root from 16.1–41.5 hr. There was no indication of polyploidy in any part of the apical meristem except in a few procambial cells. Thus, the results support the classical concept that the apical cell is the ultimate source of cells in the root.     

Effects of diurnal and bidiurnal photoperiodic treatments on mitosis in Pharbitis

Abstract

Photoperiodic effect on mitotic activity of buds from dwarf Pharbitis has been analyzed. No significant differences in mitotic activity were found in plants grown under long days or diurnal (24 h) light break photoperiodic treatments. Differences in both mitotic activity and flowering were seen in plants subjected to diurnal short days, bidiurnal (48 h) short days, or bidiurnal short days with light breaks. An elevation of mitotic activity occurs in plants grown in bidiurnal photoperiodic treatments compared to diurnal treatments. The differences in mitotic activity of buds, both vegetative and floral, seem to indicate that both phytochrome and light effect on an endogenous rhythm influence meristematic activity. Also, the extended dark period of a bidiurnal short day enhances both mitosis and flowering.     

New rhythmic changes in mitosis and growth in low differentiated green and red marine macroalgae

The cell division and vegetative growth of the thalli of simply differentiated macroalgae with a diffuse growth type—Ulva pseudocurvata (Chlorophyta) and Porphyra umbilicalis (Rhodophyta)-have been studied under natural and laboratory conditions. For this purpose the mitotic index and growth rate of algae were measured over 18 days. A diurnal rhythm of the mitotic index was revealed: the minimal mitotic index was registered in morning and daylight hours (for U. pseudocurvata 1–4%, for P. umbilicalis 0.5–2%), in the afternoon the index grew and reached its maximum 1 hour before dark (for U. pseudocurvata 12%, for P. umbilicalis 7%), then it slowly decreased during the night. In the studied algal species 2–3-and 6-day rhythms of mitotic index and growth rate were found for the first time both under natural and laboratory conditions. With constant white light these rhythms persisted for 9 days, this confirms the endogenous regulation of these rhythmic variations.     

Studies on the diurnal rhythm of mevalonate metabolism by sterol and nonsterol pathways and of mevalonate-activating enzymes

Both in vivo and in vitro incorporation of mevalonic acid into nonsaponifiable lipids by 17-day-old chick liver and kidney did not show diurnal rhythm. Using 14CO2 production from MVA as an index of the shunt pathway not leading to sterols, we have demonstrated for the first time that there is no diurnal rhythm in this pathway. No significant differences were found in the specific activities of mevalonate kinase, mevalonate-5-phosphate kinase and mevalonate-5-pyrophosphate decarboxylase from chick liver and kidney throughout a period of 24 hr, using [1-14C]mevalonate as substrate. The absence of diurnal rhythm in the decarboxylase activity was corroborated by further experiments carried out using [2-14C]mevalonate-5-pyrophosphate as specific substrate of this enzyme.     

Diurnal nitrogen excretion rhythm of the functionally ureogenic gobiid fish Mugilogobius abei

This study was performed to determine the daily periodicity of urea excretion in the ureogenic gobiid fish Mugilogobius abei. In 20% seawater, urea excretion of all the fish examined showed daily periodic changes under a 12-h light-dark cycle, and some showed a free-running rhythm under constant darkness. This is the first report of a circadian rhythm in urea excretion in fishes. Daily variations in urea excretion under light-dark cycles were also observed under various conditions, i.e. exposure to water ammonia, confinement/non-confinement and solitary/group. Due to the daily variations in urea excretion, urea contents in tissues changed periodically, whereas enzyme activities related to urea synthesis did not change significantly. The index of urea permeability as determined by changes in body urea contents after 2-h immersion of 25 mM urea solution was high during the peak of daily variation in urea excretion. Locomotor activity and urea excretion showed clear daily variations under light-dark cycles, both of which were diurnal. Furthermore, daily variations in urea excretion were maintained even when the diurnal pattern in the locomotor activity was disturbed. These results suggest that periodic urea excretion was mediated by periodic enhancement of permeability for urea at excretion sites.     

The circadian variations in the epithelial growth of the hamster cheek pouch: quantitative analysis of DNA distributions

The pronounced diurnal rhythm in DNA distributions of the hamster cheek pouch epithelium both in the S fraction and in the (G2 + M) fraction was compared with previous studies of the changes in tritiated thymidine labelling index and mitotic activity. The DNA distributions were obtained by flow cytometry after ultrasonic disaggregation of the isolated epithelium into a suspension of single nuclei. The DNA distributions were analysed with the computer program of J. Fried (1976) and by planimetry. The S fraction was higher than the autoradiographic labelling index during the whole 24 hr period. Only the computer fitted S fraction and the labelling index had the same difference between maximal and minimal values, and maxima at the same time of day. The DNA distributions showed a diurnal release of G1 cells into S phase proceeding through (G2 + M) phase and returning to G1 phase within a 24 hr period.     

Epidermal cell kinetics of the pig: a review

Age-related changes in cell kinetic parameters for the epidermis of pigs have been shown to be small, indicating that young pigs may be used for experimental studies. It was not possible to draw any firm conclusions about any strain-related differences in the cell kinetics of the epidermis of the pig. Lower LI values have been quoted for the miniature pig and the Yorkshire pig than for the Large White pig. However, these differences may be related to variations in experimental technique. The cell kinetic data for the Yorkshire pig are not consistent. Very high values for the mitotic index suggested a high rate of cell turnover, whilst data from single pulse labelling and grain count halving studies indicate a relatively low rate of cell turnover. The results from continuous labelling studies on the epidermis of the Yorkshire pig suggest that the basal cell turnover time (TT) is a factor of two or more shorter (136 h) than the estimates obtained using other methods. In the Large White pig estimates of TT were similar using a variety of techniques and were comparable with the TT estimate for the Yorkshire pig obtained using the continuous labelling method. There is some degree of inconsistency in the literature with regard to possible diurnal variations in the cell kinetic parameters for the epidermis. In the study of Archambeau & Bennet (1984) distinct diurnal variations were found in the LI, although the reliability of this finding is questionable due to the small number of animals used. Later studies by Morris et al. (1987) have suggested that diurnal variations are negligible in the epidermis of the pig. The majority of labelled cells (80%) in pig epidermis are located in the basal layer, although a significant proportion (20%) occurs suprabasally, in the cell layer immediately above the basal layer. Therefore, the epidermis can be regarded as having a bilayered proliferative cell compartment. The results from studies on irradiated pig skin (Morris & Hopewell, 1986, 1988, 1989) are not consistent with the presence of a homogeneous proliferative compartment in the epidermis, and are best explained by the occurrence of an heterogeneous proliferative compartment consisting of a stem cell subpopulation and a much larger population of transit proliferative cells.     

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.     

Diurnal rhythm of changes in the DNA-synthesizing cell count and the mitotic activity of the cells of jejunal epithelium under conditions of subdiaphragmatic vagotomy

Bilateral subdiaphragmatic vagotomy in rats 7 days after surgery results in increasing proliferative activity of the jejunum epithelium not changing circadian rhythm of the mitotic process. Vagotomy induces changes in the diurnal rhythm structure of DNA-synthesis in the jejunum epithelial cells, i.e. monoapical rhythm in the norm becomes biapical.     

Seasonal variation of body weight loss after bariatric surgery

Seasonal variations have been described in humans in several variables such as sleep, mood, appetite, food preferences, or body weight. We hypothesized that these variations could also influence the decrease in body weight rate in patients submitted to body weight loss interventions. Thus, here we tested the variations of weight loss according to the time of the year the surgery took place in a group patients (n = 1322) submitted to bariatric surgery in the Hospital Universitari de la Vall d’Hebron in Barcelona (geographical coordinates: 41°25?41″N 2°8?32″E). For the analysis, the percentage of total body weight loss (%TWL), excess body weight loss (%EWL) and percentage of body mass index loss (%BMIL) were calculated at 3 (n = 1255), 6 (n = 1172), 9 (n = 1002), and 12 months (n = 1076) after surgery. For %EWL and %BMIL a statistically significant seasonal variation was detected when the variables were calculated at 3 months, but not at the other times, with more weight loss in summer-fall. However, seasonal variations were not detected for %TWL (p = 0.09). The mean amplitude of the seasonal rhythm for %EWL was of 1.8%, while for the rhythm of %BMIL was 0.7%. Moreover, a second peak was detected in January–February modulating the seasonal rhythm of the two variables. Results confirm seasonal variations in humans and indicate that short term responses to weight loss can be modulated by the time of year.     

The Circadian Variations In the Eithelial Growth of the Hamster Cheek Pouch: Quantitative Analysis of Dna Distributions*

The pronounced diurnal rhythm in DNA distribution of the hamster check pouch epithelium both in the S fraction and in the (G₂+ M) fraction was compared with previous studies of the changes in tritiated thymidine labelling index and mitotic activity. the DNA distributions were obtained by flow cytometry after ultrasonic disaggregation of the isolated epithelium into a suspension of single nuclei. the DNA distributions were analysed with the computer program of J. Fried (1976) and by planimetry. the S fraction was higher than the autoradiographic labelling index during the whole 24 hr period. Only the computer fitted S fraction and the labelling index had the same difference between maximal and minimal values, and maxima at the same time of day. the DNA distributions showed a diurnal release of G₁ cells into S phase proceeding through (G₂+ M) phase and returning to G₁ phase within a 24 hr period.     

Mie-type scattering and non-Beer-Lambert absorption behavior of human cells in infrared microspectroscopy

We report infrared microspectral features of nuclei in a completely inactive and contracted (pyknotic) state, and of nuclei of actively dividing cells. For pyknotic nuclei, the very high local concentration of DNA leads to opaqueness of the chromatin and, consequently, the absence of DNA signals in the IR spectra of very small nuclei. However, these nuclei can be detected by their scattering properties, which can be described by the Mie theory of scattering from dielectric spheres. This scattering depends on the size of the nucleus; consequently, quite different scattering cross-sections are calculated and observed for pyknotic and mitotic nuclei.     

Diurnal Variations of Striatal D2 Dopaminergic Receptors and its Relation with Haloperidol-induced Catalepsy

In spite of the clear evidences for the blockade of dopaminergic D2 receptors as the mechanism of action for haloperidol-induced catalepsy, the contribution of pharmacokinetic and pharmacodynamic aspects on the diurnal modulation of haloperidol-induced catalepsy is controversial. We studied the diurnal variations of striatal dopamine receptors and its relation with catalepsy diurnal variations. The [3 H]-spiperone binding to dopamine receptors had a clear rhythm with a peak at 00:00 to 03:00 h, and a trough at 12:00 to 18:00 h. Haloperidol-produced catalepsy measured with the four-cork test, also showed a clear rhythm, with a peak at 00:00 h and trough at 9:00 h. The dose-response curves at peak and trough of catalepsy had the same ED 50 (0.12 mg), with time-related changes in the maximal effect. Similar diurnal variations between catalepsy and dopamine receptor binding, indicate a relevant role of temporal pharmacodynamics of haloperidol on the modulation of its behavioral effects.     

Diurnal Variations of Striatal D2 Dopaminergic Receptors and its Relation with Haloperidol-induced Catalepsy

In spite of the clear evidences for the blockade of dopaminergic D2 receptors as the mechanism of action for haloperidol-induced catalepsy, the contribution of pharmacokinetic and pharmacodynamic aspects on the diurnal modulation of haloperidol-induced catalepsy is controversial. We studied the diurnal variations of striatal dopamine receptors and its relation with catalepsy diurnal variations. The [ 3 H]-spiperone binding to dopamine receptors had a clear rhythm with a peak at 00:00 to 03:00 h, and a trough at 12:00 to 18:00 h. Haloperidol-produced catalepsy measured with the four-cork test, also showed a clear rhythm, with a peak at 00:00 h and trough at 9:00 h. The dose-response curves at peak and trough of catalepsy had the same ED 50 (0.12 mg), with time-related changes in the maximal effect. Similar diurnal variations between catalepsy and dopamine receptor binding, indicate a relevant role of temporal pharmacodynamics of haloperidol on the modulation of its behavioral effects.