Spatial distribution and cell kinetics of the glands in the human esophageal mucosa

The glands in the human esophageal mucosa have been considered rudimentary, and have been poorly studied. Only recently, their role in the defense of the esophageal mucosa in gastroesophageal reflux disease has been put in evidence. In the present study, the presence of the different esophageal gland types was observed in 82 necropsy specimens. A precise topographical study was possible in 15 specimens. Cell proliferation parameters, namely DNA synthesis and mitotic index values in the squamous epithelium and in the esophageal glands were measured in 8 patients undergoing a surgical palliative esophagectomy, after in vivo labelling with bromodeoxyuridine. Upper mucosal, lower mucosal, and submucosal glands were observed in respectively 3.7%, 87% and 99% of the 82 esophageal specimens. Our topographical analysis showed that the upper mucosal glands, lower mucosal glands, and submucosal glands are occupying respectively 0.02%, 2.61%, and 3.96% of the esophageal surface. DNA synthesis and mitotic index values in the progenitor zone of the squamous epithelium were respectively 7.91% and 0.81%, whereas the proliferative activity in the glands was extremely low. The labelling index In the excretory ducts of the glands was only 0.07%, while no labelled cells nor mitotic figures were observed in any of the glandular acini.     

AUTOSYNCHRONIZATION OF RAT LIVER CELLS WITH ENDOGENOUS CORTICOSTERONE AFTER PARTIAL HEPATECTOMY

After adrenalectomy in adult male rats ³H-TdR incorporation into the liver parenchymal cells is increased 4–8 times and the mitotic index rises from 0–31 % to 1–3%; this is inhibited by corticosterone. After hepatectomy the serum corticosterone level increases from 18 μg/100 ml to 57 μg/100 ml. The corticosterone binding capacity of the serum declines from 2–06 to 0–17. The activity of tyrosine transaminase doubles, whereas the incorporation of ³H-TdR into the liver cells is decreased by a factor of 5–7. Thereafter the binding capacity increases again and reaches, 24 hr after operation, a value of 3–82. The tyrosine transaminase activity and the serum corticosterone content return to normal. ³H-TdR incorporation, however, increases by a factor of 7-7 of the initial value. We concluded that in the first few hours after partial hepatectomy corticosterone blocks the liver cells in G₁ and an accumulation of the cells occurs at this cell cycle phase. Folio wing the binding of the corticosterone by serum proteins a little later the liver cells enter the S-phase synchronously.     

Proliferation patterns in the canine endometrium during the estrous cycle

The proliferative activity in the endometrium of 58 bitches in different stages of the estrous cycle was assessed by immunohistochemical detection of the Ki-67 proliferation associated nuclear antigen and by counting mitotic figures. The Ki-67 labelling index and the mitotic index were determined in the surface epithelium, the stroma, the crypts and the basal glands by calculating the percentage of Ki-67 positive cells and mitotic figures, respectively, on a total of 500 cells of each category. Endometrial vascular proliferation was also verified by counting the number of Ki-67 positive cells on a total of 100 endothelial cells. The present study showed two proliferation peaks involving different cell groups. In the surface epithelium, the stroma, the blood vessels and the crypts, the highest labelling and mitotic indexes were noticed during proestrus, whereas for the basal glands these indexes significantly increased (P < 0.05) during estrus compared to late metestrus and anestrus. Furthermore, a slightly positive correlation (P < 0.05) was found between the labelling index in the basal glands and the serum progesterone levels, whereas the labelling indexes in the other cell groups were positively correlated with the estradiol-17 beta levels, although not always significantly. These findings suggest that regulation of the proliferation in the surface epithelium, the stroma, the blood vessels and the crypts is different from the proliferation in the basal glands.     

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.     

Postprandial cell proliferation in the esophageal epithelium of rats

Food intake is known to trigger cell growth in the mucosa of several gut segments. In this study, the effects of both oral feeding and intragastric feeding on cell proliferation in the esophageal epithelium of rats were examined. A similar study was carried out in antrectomized animals. Refeeding of fasted rats either orally or through an intragastric catheter increased the esophageal epithelial labeling index (LI) 310 and 445%, respectively, while the mitotic index (MI) increased 427 and 217%, respectively. Under the same experimental settings, the serum gastrin values increased 423 and 200%, respectively. After surgical resection of the antrum, a postprandial proliferative wave was still observed in the orally fed rats, with an increase in LI and MI of 114 and 166%, but not in the animals refed through the gastrostomy. This study demonstrates the growth stimulating effect of feeding on the rat esophageal epithelium. This effect appears to be triggered by the mechanical passage of food and the antral release of a systemic factor, which is most probably gastrin.     

The reactivity of the hypophyseal-adrenal system in rats with different forms of adaptive behavioral pathology]

The dexamethasone suppression test (DST) was applied to male Wistar rats with different models of depression: group with the learned helplessness, group with informational neurosis provided by time-deficit conditioned avoidance training, as well as groups of rats of two strains selected for low (KLA)--and high (KHA) avoidance learning. The pre-dexamethasone basal and stress-induced corticosterone levels were similar in intact rats and those exposed to inescapable shock. The dexamethasone administration (5 mkg/kg) failed to decrease the serum corticosterone level in rats with learned helplessness. The informational neurosis increased significantly the basal corticosterone level and decreased the stress response. Serum corticosterone levels were similar in KLA and KHA rats. These results give evidence that two stress-induced rat models of depression with similar behavioural disturbances (reduction of escape/avoidance reactions) exhibit marked differences in the activity of hypothalamo-pituitary-adrenal (HPA) axis.     

Influence of prenatal stress on the rat hypothalamic-pituitary-adrenal axis activity: the role of the brain glycocorticoid receptors

The effects of prenatal stress on the hypothalamic-pituitary-adrenal (HPA) axis activity and brain glycocorticoid receptors were studied in neonatal male and female offspring, as well as the influence of neonatal glycocorticoid receptors blockade on hormonal stress reactivity of adult rats. The results showed that there were sexual differences in plasma corticosterone level and corticosteroid binding in the cortex and hypothalamus of 5-day old control rats. Prenatal stress increased basal level of corticosterone in female rats, decreased corticosterone binding in hypothalamus and hippocampus of male and female rats, and increased corticosteroid receptor level in the male cortex. Neonatal administration of glycocorticoid receptor antagonist did not change plasma corticosterone level in 5-day old rats, but prolonged hormonal stress response of the HPA axis in adult male rats and increased hormonal stress response in female ones. The character of the IIPA axis activity of male and female rats with neonatal blockade of glycocorticoid receptors correspond to hormonal stress response of prenatal stressed rats. These data suggest that change of brain glycocorticoid receptors function in neonatal period of development might be one of the mechanisms of prenatal stress influence on the HPA axis activity in the adulthood.     

Cell proliferation and growth-associated protein 43 expression in the olfactory epithelium in Poecilia reticulata after copper solution exposure

This study investigated the regeneration in the olfactory mucosa of the teleostean fish Poecilia reticulata when returned to dechlorinated tap water after 4-day exposure to 30 microg/L of Cu(2+). The regeneration process in the olfactory tissue was examined in fishes at 0, 3, 6 and 10 days of recovery in well water. Jade B staining permitted to evaluate the rate of the damage which was especially extended to olfactory neurons. Immediately after the end of exposure, a massive mitotic activity in the basal region of the mucosa was detected by immunostaining with PCNA. After 3 days of recovery the nuclei of the newly formed cells had already finished their migration to the upper portion of the epithelium, and cellular division was much less intense. Simultaneously, immunoreactivity for the neural growth-associated phosphoprotein GAP-43 increased respect to control levels, revealing that the new differentiating PCNA-positive elements belonged to immature neurons. After 6 days in well water no mitotic activity was detected, while the GAP-43 labelling appeared particularly concentrated in the apical surface of the olfactory epithelium. After 10 days the aspect of the olfactory epithelium was almost identical to the control. The present results suggest that after 10 days regeneration seems to be complete and integrity of the tissue restored. Furthermore, the epithelium reconstitution does not show apparent divergence from other fishes or mammals.     

Cytostatic effect of cyclophosphane on esophageal tumors and epithelium of mice

Investigation of cytostatic activity of cyclophosphamide in sarcoma 37 and esophagus epithelium in albino mice with respect to the diurnal rhythm of mitotic activity and the number of labeled nuclei was performed. Apparently the tumour cells in the G1-phase and at the beginning of the S-phase of the mitotic cycle were the most sensitive to the inhibitory effect of this drug. During the completion of the DNA-synthesis period the cell resistance to the action of the cytostatic increased. Cells at the G1-phase of the mitotic cycle were sensitive to the inhibitory action of cyclophosphane in the esophageal epithelium.     

Effects of galanin on the secretion and proliferative activity of the immature and regenerating adrenal glands of rats

The effects of galanin and the galanin-receptor antagonist (galanin-A) [D-Thr(6),D-Trp(8,9),15-ol]-galanin(1-15) on the immature and regenerating rat adrenal glands have been investigated in vivo. Adult female rats with adrenal regeneration and their offpring (20-day-old) were given three subcutaneous injections (28, 16, and 4 h before being killed) of 2 nmol/100 g galanin and/or galanin-A, and 0.1 mg/100 g vincristin 3 h before being killed. Plasma corticosterone concentration was measured by radioimmunoassay, and the mitotic index ( per thousand of metaphase-arrested cells) was evaluated. In immature rats, galanin increased plasma corticosterone concentration, without affecting mitotic index; the secretagogue effect was reversed by galanin-A, which alone was ineffective. In rats with regenerating adrenal, galanin-A increased both blood level of corticosterone and mitotic index; galanin was ineffective, but blocked the effects of galanin-A. These findings allowed us to draw the following conclusions: 1) galanin exerts a moderate glucocorticoid secretagogue action on immature rat adrenals, but endogenous galanin does not play a major physiological role in the functional control of the gland; and 2) endogenous galanin exerts a maximal tonic inhibitory control on both glucocorticoid secretion and proliferative activity of regenerating rat adrenals, whose physiological relevance remains to be investigated.     

Cyclic AMP and serum arrest of the mitotic activity of human diploid fibroblasts.

Mitotic activity in confluent cultures of human diploid fibroblasts was arrested by the reduction of the serum concentration of the incubation medium to 0.5% or by the addition of 0.5 mM 6-N, 2'-O-dibutyryl-adenosine 3':5'-cyclic monophosphate (db cAMP). Under either of these conditions, cultures maintained a constant cell number for 14 days; cultures continuously exposed to medium containing 10% serum doubled their cell number during this 14-day period. The protein cotent per cell decreased by 20% when cells were maintained with 0.5% serum whereas that of cells exposed to db cAMP remained constant. Ultrastructural studies revealed that cells exposed to db cAMP exhibited a morphology typical of cells cultures with 10% serum alone, whereas cells incubated with 0.5% serum showed the ultrastructural changes in mitochondria, endoplasmic reticulum and Golgi complex previously identified with low-serum arrest. Cellular adenosine 3':5'-cyclic monophosphate (cAMP) levels remained constant during the 7-day growth period in which confluency was attained, as well as during the 14-day arrested period with 0.5% serum. These results indicated that the mitotic inhibition induced by reducing the serum concentration of the incubation medium was not mediated by increased intracellular levels of cAMP and differed from that induced by the addition of exogenous db cAMP.     

The neonatal glucocorticoid treatment-produced long-term changes of the pituitary-adrenal function and brain corticosteroid receptors in rats.

Two distinct periods of sensitivity to elevated glucocorticoid hormone levels during postnatal development of the pituitary-adrenal axis were studied. Wistar rats were injected subcutaneously (s.c.) with cortisol (1 mg/kg) on postnatal days 1-5 or 14-18. The steroid treatment during the first postnatal week resulted in a decrease of the morning basal and stress-induced plasma corticosterone levels in 30 day-old male rats, as well as in rats that were injected with cortisol on the third postnatal week. Stress-induced corticosterone levels in 90-day old cortisol-treated rats were determined in blood samples drawn from the tail vein before the restraint stress, immediately after the 20-min long stress, then 60 and 180 min afterwards. Only the rats treated with cortisol during the third week showed a prolonged stress-induced corticosterone secretion, with the highest corticosterone level in 180 min after the restraint stress. The early neonatal cortisol treatment had no effect on (3)H-corticosterone binding in all studied brain areas of the 90-day old rats. The rats treated with cortisol at the 14-17th postnatal days showed a significantly lower (3)H-corticosterone binding in the frontal cortex, hippocampus, and hypothalamus. These findings suggest that the third week of life in rats is more sensitive to elevated levels of corticosterone than the first one. The high level of glucocorticoids at this period has long-term effects on the efficiency of the negative feedback mechanisms provided by hypothalamus-pituitary-adrenal axis.     

Cellular necrosis in zinc-deficient rat embryos

Abnormal cellular necrosis was studied in 9.5-11.5-day embryos obtained from zinc-deficient rats. At periods of low maternal zinc status induced by a high intake of a zinc-deficient diet, cell death was observed in those regions of the embryo that were most sensitive to teratogenic insult at that time. As the maternal serum zinc level increased during the fasting phase of the feeding cycle, the degree of necrosis decreased, leaving the tissue histologically more normal even though the embryos were grossly malformed. The mitotic index of cells in the neural epithelium and limb buds of zinc-deficient, non-necrotic embryos was found to be elevated, but there was no evidence of blockage at any particular stage of mitosis. It can be hypothesised that during the early stages of organogenesis, periods of low maternal zinc status initiate unscheduled cell death by some as yet undefined mechanism that in turn, gives rise to the morphological anomalies observed later.     

Circadian synchronization of hepatocyte proliferation in young rats: the role played by adrenal hormones

The circadian rhythm of hepatic cell proliferation in rats appears on the 20th day of life, when the hypothalamo-adrenal axis is mature enough for circadian activity to occur. From the 20th day to the 30th day of life, the mitotic rhythm is progressively induced by a reduction in nocturnal values, while diurnal rhythms remain unchanged. Mitotic peaks emerge at 10.00 hours. A labelling index wave occurs 8 hr before the corresponding mitotic wave, with a peak at 02.00 hours and a minimum in the evening, coincidental with the acrophase of plasma corticosterone level (activity phase). Labelled mitoses curves and metaphase accumulation after colchicine injection show that the duration of the S, G2 and M phases remain approximately constant and that the circadian variation is due to a variation in the rate of cells that enter these successive phases. During the synchronization period (from day 20 to 30), the growth fraction decreases progressively. Adrenalectomy at this time is followed by a higher cell proliferation and all rhythms disappear after 2 days. Corticosterone injected before the triggering of the rhythmic activity in 17-day-old rats immediately reduces the labelling index, while the mitotic index is decreased 10 hr later; this delay is equal to the S + G2 duration. The results are discussed. They favour the hypothesis that the circadian variation of corticosterone is responsible for the induction of a circadian variation in developmental cell proliferation by inhibition of the G1-S transition when it is higher in the evening.     

Destabilizing effect of chicken selection using the functional adrenal reserves criteria]

Chicken lines produced by divergent selection for the functional adrenal reserves showed significant between-line differences in the content of corticosterone and other hormones (thyroxin, progesterone), as well as in body weight, early maturation, and egg yield. DNA fingerprinting with the pGB725 probe revealed molecular changes in genomic DNA of the chicken lines subjected to plus and minus selection. The genetic distances between the original population and the selected chicken lines, which were estimated from the molecular hybridization patterns, reflected the history of breeding. Analysis of mixed DNA from several individuals of each line revealed specific hybridization bands that could serve as DNA markers during selection for the high and low corticosterone levels in blood.     

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.     

Destabilizing Effect of Chicken Selection for the Functional Adrenal Reserves

Chicken lines produced by divergent selection for the functional adrenal reserves showed significant between-line differences in the content of corticosterone and other hormones (thyroxin, progesterone), as well as in body weight, early maturation, and egg yield. DNA fingerprinting with the pGB725 probe revealed molecular changes in genomic DNA of the chicken lines subjected to plus and minus selection. The genetic distances between the original population and the selected chicken lines, which were estimated from the molecular hybridization patterns, reflected the history of breeding. Analysis of mixed DNA from several individuals of each line revealed specific hybridization bands that could serve as DNA markers during selection for the high and low corticosterone levels in blood.     

Radioautographic study of the cell proliferation of the pigment epithelium of the retina in albino clawed frogs

The proliferative activity of pigment epithelium was studied by means of 3H-thymidine autoradiography after the removal of retina, lens and iris with the ciliary-terminal zone in the adults. The cell population of pigment epithelium was shown to be heterogeneous on the level of proliferative activity. A low level of proliferation is characteristic of the cells of epithelial monolayer and the cells leaving it and forming aggregates. An intensive local proliferation leading to the formation of expansions was found in the pigment epithelium layer in 7% of cases. On the 20th day after the operation, the index of labelled nuclei in the expansions amounted to 43.4--59.3% and the mitotic index to 1.4--2.1%. On the 75th day elements of atypical retinal differentiation, besides the high proliferative activity, were observed in one expansion.     

The cell kinetics of the adaptation of the human esophagus to organ culture

Summary The adaptation of normal human esophageal explants to organ culture for the first 33 d of in vitro growth was evaluated using histomorphology and [³H]TdR autoradiography combined with mitotic blockade. On the 3rd d in culture, extensive desquamation of superficial cells reduced the epithelium to about four cell layers. Thereafter, the epithelium remained atrophic, with a relative increase in basal and suprabasal cells. The percentage of cells synthesizing DNA was greatest from Day 4 through 8, just after desquamation, and reached a maximum on Day 4 (24 h [³H]TdR labeling index of 62%). The labeling index (LI) fluctuated, thereafter, but remained high (26% on Day 33). During the last 6 h of each [³H]TdR labeling interval, mitosis was blocked by colcemid. The 6 h mitotic rate (MR) was a reasonably constant fraction of the LI (maximum at 4 d: MR=1.44%), but was much lower than predicted by [³H]TdR labeling indicating the loss of large numbers of cells after DNA synthesis but before or during mitosis. Unlabeled mitotic figures appeared between Days 1 to 3 and 6 to 33, suggesting that the epithelium initially contained a considerable population of cells arrested or delayed in G₂ and continued to generate cells that remained in premitosis longer than 24 h. These results indicate that the atrophy observed in vitro is characterized by a relative increase in the basal and suprabasal cell category, a high replication rate, initial recruitment of cells arrested in premitosis, and rapid cell turnover with significant loss of cells at the premitotic or mitotic step, or both. Thus it seems that human esophageal epithelium grown in organ culture is a satisfactory substrate for experimentation (for example, in vitro carcinogenesis) that requires cell replication. However, there are major differences between the kinetics of esophageal epithelium in vivo and in vitro. Supported in part by Contract NOI-CP-75909 and NOI-CP-25604-59 from the National Cancer Institute, Bethesda, MD.     

Mitotic activity of the lymphocytes of the thymus cortex in hypokinesia during the period of readaptation

The changes in the weight and mitotic index were studied in the cortex of the thymus of Wistar rats during 10-day hypokinesia and 10-day readaptation (restoration). 24 hours after immobilization of the animals the mitotic index was 2 times as lower. No complete readaptation was attained during 10-day hypokinesia. No readaptation was attained during 10-day hypokinesia. In readaptation the stage of secondary stress was found (the mitotic index was 3.5 times as reduced), the stage of genuine restoration being revealed after 10 days.