Non-circadian periodicity in the duration of the cell cycle and the G1 phase in the hindlimb epidermis of the anuran tadpole, Rana pipiens

Using the percentage labeled mitoses method, seven cell cycle determinations were initiated at 6-hr intervals over a 36-hr span in order to see if the cell cycle in the tadpole hindlimb epidermis varied with time or showed rhythmicity. There was a pattern of two long cell cycles followed by a shorter one. Total cell cycle length (Tc) and the length of the G1 phase plus one-half of the mitotic time (TG1 + 1/2M) fluctuated the most, although only TG1 + 1/2M varied significantly with the Chi-square test. The proportion of TC spent in each phase was also calculated. Only TG1 + 1/2M/TC had statistically significant fluctuations with time. Rhythmicity was analyzed by a computer program using the method of least squares for cosine curve fitting. Statistically significant ultradian rhythms of 18.4 hr in TC, 18.5 hr in TG1 + 1/2M and 18.6 hr in TG1 + 1/2M/TC and the length of the DNA synthetic phase/total cell cycle length (TS/TC) were found. Circadian rhythmicity was not observed. The acrophases of the ultradian rhythms of TC and TG1 + 1/2M coincided, suggesting that the rhythm of TC was due mainly to variation in TG1 + 1/2M. In the absence of significant variation in TS, the longest phase of the cell cycle, whenever G1 + 1/2M was short, TS/TC increased, so that the 18.6 hr rhythm in TS/TC was also a result of the periodicity in TG1 + 1/2M.     

Axillary temperature rhythms: the predominance of ultradian periodicity in major affective disorders

Axillary temperature was recorded at least twice during a 48 hrs. span at 6 min. intervals in 10 hospitalized subjects with major affective disorders (DSM III 296. xx). During the clinical occurrence of acute symptoms 7 out of 10 subjects exhibited a prominent ultradian periodicity (period tau less than 20 hrs.) in their temperature time series. Whatever the used therapeutic mean (electroconvulsive therapy and/or chemotherapy) the improvement was associated with a circadian rhythmicity (20 hrs. less than or equal to tau less than or equal to 28 hrs.). A prominent temperature ultradian rhythm (which occurs only in the new born) could be the index of an internal desynchronization associated with major affective disorders.     

The antimitotic effect of the neem terpenoid azadirachtin on cultured insect cells

When cultured insect cells (Sf9) were grown in the presence of 5 x 10(-6) M azadirachtin, there was a rapid increase in the mitotic index, with the appearance of many aberrant mitotic figures. Flow cytometry established that cells accumulated in the G2/M phase of the cell cycle, and that the effect was concentration-dependent. At 10(-8) M a period of 20 h was necessary to raise the proportion in G2/M to 42% above the control values, but at 5 x 10(-6) M more than 90% of the cells were in this phase. Azadirachtin had the same effect on C6/36 mosquito cells, but failed to affect L929 murine fibroblast cells even at a concentration of 10(-4) M over 72 h. Experiments with colchcine and taxol showed similarities of action between azadirachtin and colchicine, and azadirachtin was apparently able to displace colchicine-fluorescein from binding-sites in living insect cells. Another similarity between azdirachtin and colchicine was that both phytochemicals prevented the polymerisatrion in vitro of mammalian tubulin, although the azadirachtin was much less effective.     

Plasma concentrations of thyroid hormones in dogs: influence of sampling hour, breed and age

Thyroxine (T4) and triiodothyronine (T3) plasma concentrations have been determined during 24-hr sampling periods in six mongrels (age 12-36 months), six beagles (age 35-37 months), three labradors (age 3.5 months) and three beagles (age 5 months). The mean T4 levels of the labradors were significantly lower than the values found for mongrels or older beagles (P less than 0.05), whereas T3 was higher in the 5 month old beagles compared to the mongrels (P less than 0.001), young beagles (P less than 0.05) or labradors (P less than 0.01). Circadian and ultradian rhythmicities have been evaluated by cosinor and Fourier analysis. Mongrels and older beagles did have a 12-hr rhythmicity in plasma T4 (P less than 0.05), whereas 5 month old beagles had a circadian one (P less than 0.01). A 12-hr rhythmicity was also found for T3 in the older Beagles (P less than 0.05). However, Fourier analysis indicated that the daily variation in T4 and T3 plasma levels was inadequately mathematically described by single sinusoidal rhythm and that more harmonic components are to be taken into account. The obtained data during a 24-hr period indicate that T4 and T3 concentrations in plasma may vary according to breed, age and sampling hour.     

Differential elimination of circadian and ultradian rhythmicity by hypothalamic lesions in the common vole, Microtus arvalis

Effects of hypothalamic lesions on the ultradian and circadian organization of wheel running and feeding were studied in the common vole, Microtus arvalis. Circadian organization broke down within 30 days in continuous darkness in 24% of intact voles (n = 135). Ultradian rhythmicity of feeding (period 2-3 hr) persisted in constant conditions in all intact voles. Following lesions of the suprachiasmatic nuclei (SCN), circadian rhythmicity disappeared when lesions were complete (n = 8) or more extensive than 25% of the total SCN volume (n = 5). Absence of circadian rhythmicity was also found in animals with substantial lesions in the diencephalic paraventricular area (PVA) and in the retrochiasmatic area (RCA) and/or adjacent arcuate nucleus (Arc). Complete loss of ultradian and circadian organization occurred in eight voles with damage to the RCA and/or Arc. In three of these, the SCN was intact. The SCN is a likely candidate for a circadian pacemaker in voles (as in other rodents), while the loss of circadian rhythmicity following PVA and RCA/Arc lesions may be due to destruction of efferent pathways from the SCN. The RCA/Arc area is apparently necessary for the expression of ultradian rhythms. The intact SCN is neither necessary nor sufficient for the generation of ultradian rhythmicity.     

Circadian and Ultradian Rhythmicities in Very Premature Neonates Maintained in Incubators

Six very premature babies (born at 26-28 weeks gestational age) have been studied in hospital for 11-17 weeks, while in intensive care and in an incubator. Apart from suffering occasionally from the neonatal disorders of haemolytic jaundice and 'respiratory distress of the newborn', the babies were healthy and developed normally. Initially, the babies were continuously fed intravenously, and the lighting in the ward was on continuously. Routine care was given round the clock. When their medical condition permitted it, the babies were moved in their incubator to an adjacent ward, where they took frequent (2-4 hourly) small meals by mouth, the lighting was dimmed at night, and routine care tended to be given more in the daytime. Hourly recordings of insulated skin temperature were taken throughout the study, and it is the detection of rhythmicity in these measurements that has been the subject of the present study. The methods used were Phase-weighted Stacks, Phasor Walkout and Power Spectral Analysis. These methods have previously been used mainly in geophysical studies, and their value is that they can detect weak signals in noisy data and do not assume a particular waveform of any signal. Circadian rhythmicity was found in all babies for much of the time that were in the constant environment provided by the incubator. Ultradian rhythms were sometimes present also, but they were shorter-lived, and showed a wide range of changing periods, generally in excess of 8 h. When the babies were being treated for jaundice or respiratory distress, there was a tendency for the circadian rhythms to become weaker and for a broader spectrum of ultradian periods to appear. Placing babies in the 12 h : 12 h light : dark environment provided by the ward, and instituting feeding by mouth, had, in most cases, only modest effects upon either circadian or ultradian rhythms. Thus, circadian rhythms continued (but generally with a period not exactly equal to 24 h), and ultradian rhythms, when present, often did not show periods that could be related easily to feeding or care-giving. These results are discussed in terms of evidence for endogenous and exogenous origins of the observed rhythms, and of theories that have postulated the relationship between circadian and ultradian rhythms. It is concluded that the results from the present analyses are difficult to reconcile with the view that circadian rhythms develop from interactions between ultradian oscillators. We suggest that they indicate a matu-ration of the circadian system as a consequence of increasing associations between the circadian elements that are present in the suprachiasmatic nuclei and in other oscillators of the circadian system. The new analytical methods used here also indicate that the results obtained from time-frequency analysis depend to some extent upon the method used.     

Circadian rhythmicity within single cells of Paramecium bursaria

Cell populations of Paramecium bursaria show mating reactivity in the light period, but not in the dark period, when exposed to a light-dark cycle (LD 12:12). After they are transferred to constant-light (LL) conditions (1,000 lux), they continue to show a circadian rhythm of mating reactivity. The rhythm gradually dampens in LL so that mating reactivity in populations becomes arrhythmic in LL within 2 weeks. We wanted to know whether the arrhythmicity of this population was due to the absence of circadian rhythmicity within each individual cell, or merely due to asynchrony of a population of individually rhythmic cells. Therefore, single cells were isolated randomly from an arrhythmic population that had been in LL for a long time. Then the mating reactivity of these single cells was individually tested every 3 hr for 2 days. Each single cell showed a circadian mating rhythm in LL. This shows that the disappearance of the mating rhythm in cell populations under LL is not caused by disappearance of circadian rhythmicity within individual cells, but is due to desynchronization among cells in a population. When an arrhythmic population in LL is darkened for 9 hr, the mating reactivity rhythm of the cell population reappears. This occurs by resynchronization of the rhythms among individual cells, as can be shown by exposing single cells to pulses of 9 hr of darkness. This dark treatment causes phase shifts of single-cell rhythms, and a phase response curve is obtained for this stimulus. This phase-shifting behavior explains the efficacy of 9-hr dark pulses in restoring the population's rhythm.     

Induction of structural and numerical changes of chromosome, centrosome abnormality, multipolar spindles and multipolar division in cultured Chinese hamster V79 cells by exposure to a trivalent dimethylarsenic compound

Dimethylarsine iodide (DMI) was used as a model compound of trivalent dimethylarsenicals [DMA(III)], and the biological effects were extensively investigated in cultured Chinese hamster V79 cells. When the cytotoxic effects of DMA(III) were compared with those of inorganic arsenite and dimethylarsinic acid [DMA(V)], DMA(III) was about 10,000 times more potent than DMA(V), and it was even 10 times more toxic than arsenite. Depletion of cell glutathione (GSH) did not influence the cytotoxic effects of DMA(III), whereas it enhanced the cytotoxicity of arsenite. Chromosome structural aberrations, such as gaps, breaks and pulverizations, and numerical changes, such as aneuploidy, hyper- and hypo-tetraploidy, were induced by DMA(III) in a concentration-dependent manner. Mitotic index increased 9-12h after the addition of DMA(III), and then declined. By contrast, the incidence of multinucleated cells increased conversely with the decrease in mitotic index at and after 24h of exposure. The mitotic cell-specific abnormality of centrosome integrity and multipolar spindles were induced by DMA(III) in a time- and concentration-dependent manner. Moreover, DMA(III) caused abnormal cytokinesis (multipolar division) at concentrations that were effective in causing centrosome abnormality, multipolar spindles and aneuploidy. These results showed that DMA(III) was genotoxic on cultured mammalian cells. Results also suggest that DMA(III)-induced multipolar spindles and multipolar division may be associated with the induction of aneuploidy. In addition, the centrosome may be a primary target for cell death via multinucleated cells.     

Twenty-four-hour and Ultradian Rhythmicities in Healthy Full-Term Neonates: Exogenous and Endogenous Influences

Eleven healthy, full-term babies were studied on the second day after birth and again 4 weeks later. The babies lived on a 24-h light/dark cycle (light from 0700D1900) and were bottle-fed every 4 h. Systolic blood pressure, heart rate, skin (abdomen) and rectal temperatures were measured at 10-min intervals for 24 h on each occasion of study. The behavioural state of the baby was measured at the same time, and this information was used to purify the raw data (i.e., to separate it into the endogenous, clock-driven and exogenous, lifestyle-driven components). Raw and purified data were assessed for 24-h and ultradian (12-, 8-, 6-, 4-, 3-, 2-h) periodicities by cosinor analysis. We confirm the development of 24-h rhythmicity in skin and rectal temperatures between day 2 and week 4; at the same time, ultradian rhythms (4-h) developed in all variables. For heart rate and systolic blood pressure the development of a 4-h ultradian rhythm was in phase with the behavioural changes produced by feeding; by contrast, for the temperatures, these weak exogenous components were accompanied by a stronger 4-h component, that was out of phase with feeding. Masking effects due to sleep and activity changed in size between day 2 and week 4. Also, those positive masks produced by waking activities were more marked in the light, whereas the negative masks produced by sleep were more marked in the dark. Some implications of these results for the development of rhythmicity in infants, particularly whether due to lifestyle or the development of internal processes, are discussed.     

Dna Synthetic Activity in Tumor-Bearing Mice

The rate of DNA synthesis in normal tissues exhibits circadian rhythmicity. However, there have been conflicting reports of the effects of tumor burden on the circadian rhythm of DNA synthesis in non-cancer tissues. We have developed a mouse colon cancer (MC-26) that exhibits different growth under different photoperiods. The purpose of this study was to analyze DNA synthetic activity in tissues removed from tumor-bearing and tumor-free mice maintained under two different photoperiods. Two groups each of approximately 80 male Balb/c mice were acclimated to one of two light-dark cycles, 12L:12D or 6L:18D. Half of each group were injected with 5.0 × 104 MC-26 cells. Twenty-two days later, all mice were killed in subgroups at 4-6 hr intervals over one 24-hr period. Colons and tumors were removed for measurement of DNA synthesis. Results were analyzed by means of one-way analysis of variance (ANO VA) in order to determine whether DNA synthesis varied significantly within groups over the 24-hr period. The DNA synthetic activity, as measured by uptake of tritiated thymidine, exhibited significant temporal variation in the colons of control (tumor-free) mice under both the 12L:12D and 6L:18D photoperiods. The colons of tumor-bearing mice failed to exhibit a fluctuation under a 12L:12D photoperiod but did show a significant 24-hr rhythm under the 6L:18D photoperiod. The subcutaneously growing cancers did not exhibit a circadian variation in DNA synthetic activity under either photoperiod. Both photoperiod and the presence of cancer appear to affect the DNA synthetic activity observed in mice bearing the MC-26 colon cancer.     

Multiple Component Analysis of Plasma Growth Hormone in Children with Standard and Short Stature

Growth hormone (GH) concentrations (in ng/ml) were determined by radioimmunoassay, in plasma obtained at about 3-hr intervals during a 24-hr sampling span, from 42 boys and 12 girls of short stature (2-4 standard deviations below their peer group mean), and 13 boys and 9 girls of standard stature. Subjects had 11.20 0.37 years of age at the time of study, and were living on a diurnal waking (∼07:30 to ∼22:30), nocturnal resting routine during sampling. Analysis of these data by single and population-mean cosinor methods as well as by analysis of variance revealed circadian and ultradian prominent components characterizing most groups. Accordingly, a multiple component analysis was undertaken for data of each group separately, as well as for all subjects. A comparison of circadian parameters indicates similar characteristics between short and standard children, whether one compares boys [P=0.674, 0.371 and 0.749 for comparison of rhythm adjusted means (M), amplitudes (A) and acrophases (), respectively], girls (P=0.993,0.914 and 0.397), or all children (P=0.859,0.712 and 0.865). Differences are found, however, in circasemidian characteristics as well as in the prominent 8-hr ultradian component documented for the short but not for the standard children. These ultradian components should be taken into consideration in the design and later evaluation of a time-specified treatment of children of short stature.     

Circadian rhythms in hot flashes in natural and surgically-induced menopause

The aim of this study was to investigate circadian and ultradian variations in menopausal hot flash. The number of hot flashes per 2-hr period was collected from 25 diurnally-active, perimenopausal women for 1 week in January or February of each year for 3 consecutive years. Fourteen women were experiencing natural menopause (NM) (mean age 51.9 years) and 11 were experiencing surgically-induced menopause (SIM) (mean age 52.0 years). The difference in the number of hot flashes between the two types of menopause at each clock time was not statistically significant; neither was the mean number of hot flashes per 24 hr different between the two groups (Student's t-test). Data when normalized for each woman and placed end-to-end revealed by cosinor analysis circadian rhythmicity in the SIM group (P = 0.02) but not in the NM group. A 12-hr periodicity was detected in both groups (P less than 0.001 for both). An 8-hr rhythm was detected only for the NM group (P = 0.04). Both groups combined exhibited statistically significant rhythmicities with periods of 24 hr (P = 0.003), 12 hr (P less than 0.001) and 8 hr (P = 0.005). Regardless of the type of menopause, the women could be separated into two groups based on the temporal pattern of hot flashes during the day. One group was defined by the occurrence of peak frequency of flashes during the morning (0400-0959), while the second group was defined by the occurrence of the peak in the evening (1600-2159).(ABSTRACT TRUNCATED AT 250 WORDS)     

A TEMPERATURE-COMPENSATED ULTRADIAN CLOCK OF TETRAHYMENA: OSCILLATIONS IN RESPIRATORY ACTIVITY AND CELL DIVISION

Both a circadian clock and an ultradian clock (period 4—5 h) have previously been described for the ciliated protozoon Tetrahymena. The present communication demonstrates the existence of yet another cellular clock: an ultradian rhythm with a period of about 30 min. The period was found to be well temperature-compensated over the range studied, i.e., between 19°C and 33°C. Ultradian rhythmicity was initiated by dilution of stationary-phase cultures, which were kept previously in a light-dark cycle, into fresh medium. LD treatment during stationary phase was an absolute requirement, since cultures kept in either LL or DD did not produce the ultradian rhythmicity after refeeding. The clock exerts control over respiration; the observed oscillation in oxygen uptake is just a hand of the clock: after a limitation of oxygen supply had ended, the rhythm resumed with the same phase and period as that in control cultures. The clock exerts temporal control also over cell division; in the refed culture cell division resumed with an oscillation in the number of dividing organisms. The period of this oscillation corresponded to that of the rhythm in respiratory activity, indicating that the same ultradian clock may exert control over different cellular functions. Analysis of a second Tetrahymena strain indicates that period length of the ultradian clock is a strain-specific characteristic.     

Influence of the feeding regimen on the rhythmicity of the processes of secretion of the supraoptic nucleus in C57Bl mice

During the period of vernal equinox in Leningrad 2 groups of C57Bl male mice have been investigated. Ninety-five animals are given food ecologically adequate at 9 p.m. Eighty-four animals are given foot at 9 a.m.--ecologically inverted regimen of feeding (IRF). The mice are decapitated for 4 days with an interval about 1.5 h. Serial paraffin sections are stained with aldehyde-fuchsin after Gomori and an additional staining of the nuclei with azocarmine. Criteria for the neurosecretory activity is the ratio of the cells amount at various stages of synthesis, outflux and accumulation of the secrete, volumes of the nuclei and nucleoli. Spectrum and parameters of the rhythmicity are revealed. IRF produces decrease in the amount of the ultradian component of the rhythmic parameters, characterizing active synthesis and discharge of the secrete. The part of the neurosecretory cells, those actively synthesizing and discharging the secrete, and volume of the cell nucleoli decrease. Range of ultradian component of the cell part rhythm, depositing the secrete, and the cell volume enriches. Thus, IRF produces certain changes in the rhythmicity of the cell secretion at all the stages: synthesis, discharge and accumulation of the secrete. Total intensity of the synthetic processes decreases. A conclusion is made that IRF inhibits the microsecretory process in the supraoptic nucleus (SON) and decreases adaptive possibilities of its cells. Adaptation to IRF is performed at the expense of rhythmic discharge of neurohormones, deposited in the cells, and at the expense of processes, occurring in the neuryoplasm and resulting in increase of the nuclear volume.(ABSTRACT TRUNCATED AT 250 WORDS)     

The therapeutic effect of OK-432-combined adoptive immunotherapy against liver metastases from gastric or colorectal cancers

Twenty-four patients with liver metastases from gastric or colorectal cancer were treated with OK-432-combined adoptive immunotherapy (AIT). Lymphocytes isolated from regional lymph nodes or peripheral blood were cultured with medium containing T cell growth factor and sonicated tumor extract antigen (SE-Ag) for 9–13 days. The cultured lymphocytes were transferred mainly through the hepatic artery after the administration of OK-432, a streptococcal preparation. Sixteen of the 24 patients received a low dose of anti-cancer agents between the OK-432 injection and cell transfer. When cultured without SE-Ag, regional lymph node lymphocytes (RLNL) showed significantly (P<0.05) higher cytotoxic activity against autologous tumor cells and, on the contrary, lower cytotoxic activity against K562 than peripheral blood lymphocytes (PBL). When cultured with SE-Ag, cytotoxicity of RLNL against autologous tumor cells was nearly equivalent to that of PBL. The blastogenesis of fresh PBL to SE-Ag was significantly (P<0.05) augmented after the OK-432-combined AIT. Two patients showed complete response and 4 patients showed partial response among 19 patients who had evaluable lesions. Five patients whose liver metastases were resected were treated with OK-432-combined AIT as an adjuvant therapy. To date they are alive without recurrence in the liver.Abbreviations AIT adoptive immunotherapy - RLNL regional lymph node lymphocytes - SE-Ag sonicated tumor extract antigen     

Orcadian Variation in Human Stature

Thyroxine (T₄) and triiodothyronine (T₃) plasma concentrations have been determined during 24-hr sampling periods in six mongrels (age 12-36 months), six beagles (age 35-37 months), three labradors (age 3.5 months) and three beagles (age 5 months). The mean T₄ levels of the labradors were significantly lower than the values found for mongrels or older beagles (P < 0.05), whereas T₃ was higher in the 5 month old beagles compared to the mongrels (P < 0.001), young beagles (P < 0.05) or labradors (P < 0.01).

Circadian and ultradian rhythmicities have been evaluated by cosinor and Fourier analysis. Mongrels and older beagles did have a 12-hr rhythmicity in plasma T₄ (P < 0.05), whereas 5 month old beagles had a circadian one (P < 0.01). A 12-hr rhythmicity was also found for T₃ in the older Beagles (P < 0.05). However, Fourier analysis indicated that the daily variation in T₄ and T₃ plasma levels was inadequately mathematically described by single sinusoidal rhythm and that more harmonic components are to be taken into account.

The obtained data during a 24-hr period indicate that T₄ and T₃ concentrations in plasma may vary according to breed, age and sampling hour.     

Non-Circadian Periodicity in the Duration of the Cell Cycle and the G1 Phase in the Hindlimb Epidermis of the Anuran Tadpole,Rana Pipiens

Using the percentage labeled mitoses method, seven cell cycle determinations were initiated at 6-hr intervals over a 36-hr span in order to see if the cell cycle in the tadpole hindlimb epidermis varied with time or showed rhythmicity. There was a pattern of two long cell cycles followed by a shorter one. Total cell cycle length (Tc) and the length of the G₁ phase plus one-half of the mitotic time (TG₁+½M) fluctuated the most, although only TG₁+½M varied significantly with the Chi-square test. The proportion of TC spent in each phase was also calculated. Only TG₁+½M/Tc had statistically significant fluctuations with time.

Rhythmicity was analyzed by a computer program using the method of least squares for cosine curve fitting. Statistically significant ultradian rhythms of 18.4 hr in TC, 18.5 hr in TG₁+½M and 18.6 hr in TG₁+½M/TC and the length of the DNA synthetic phase/total cell cycle length (TS/TC) were found. Circadian rhythmicity was not observed. The acrophases of the ultradian rhythms of TC and TG₁+½M coincided, suggesting that the rhythm of TC was due mainly to variation in TG₁+½M. In the absence of significant variation in TS, the longest phase of the cell cycle, whenever G₁+½M was short, TS/TC increased, so that the 18.6 hr rhythm in TS/TC was also a result of the periodicity in TG₁+½M.     

Time Periods in the Nasal Cycle During Night Sleep

The periodic congestion and decongestion of nasal venous sinuses and the alternation of airflow from one side of the nose to the other are referred to as a 'nasal cycle' in the literature. The aim of this study was to detect the nasal cycle during sleep in normal subjects and describe existing time periods and their sequence and patterns. We studied 58 records of the nasal cycle over 6-9 hours of sleep in six healthy volunteers and revealed that the cycle could be described as a combination of 1 to 4 discrete ultradian periods with various length: 1.0-1.5 h (mean 78.6min), 2.5-3.0 h (168.3 min), 4.0-4.5 h (260.3 min) or 5.5-6.0 h (347.5 min). The distribution of the discrete time periods was multi-modal and the mean lengths of periods were 'multiples' of a basic period of 85.4 min (~1.5 h) which was very close to the mean length of the sleep cycle (~1.5 h). In all subjects, during any of the REM stages of the sleep, an alternation of the airflow through the nostrils was observed. In about 75% of all cases, the switch of the flow between the nostrils occurred during the second or following REM stages of the sleep thus shaping a nasal cycle that contained mainly periods of 3.0 or 4.5 hours. We suggest a novel classification of the nocturnal nasal cyclicity and hypothesis that there is a relationship between the nasal cycle and the sleep cycle which, like other cyclic physiological phenomena with ultradian rhythmicity, expresses a pattern of 'lateralisation' that is synchronous with changes in the sleep cycle.     

Circadian and Ultradian Rhythmicities in Very Premature Neonates Maintained in Incubators

Six very premature babies (born at 26–28 weeks gestational age) have been studied in hospital for 11–17 weeks, while in intensive care and in an incubator. Apart from suffering occasionally from the neonatal disorders of haemolytic jaundice and ‘respiratory distress of the newborn’, the babies were healthy and developed normally. Initially, the babies were continuously fed intravenously, and the lighting in the ward was on continuously. Routine care was given round the clock. When their medical condition permitted it, the babies were moved in their incubator to an adjacent ward, where they took frequent (2–4 hourly) small meals by mouth, the lighting was dimmed at night, and routine care tended to be given more in the daytime. Hourly recordings of insulated skin temperature were taken throughout the study, and it is the detection of rhythmicity in these measurements that has been the subject of the present study. The methods used were Phase-weighted Stacks, Phasor Walkout and Power Spectral Analysis. These methods have previously been used mainly in geophysical studies, and their value is that they can detect weak signals in noisy data and do not assume a particular waveform of any signal. Circadian rhythmicity was found in all babies for much of the time that were in the constant environment provided by the incubator. Ultradian rhythms were sometimes present also, but they were shorter-lived, and showed a wide range of changing periods, generally in excess of 8 h. When the babies were being treated for jaundice or respiratory distress, there was a tendency for the circadian rhythms to become weaker and for a broader spectrum of ultradian periods to appear. Placing babies in the 12 h : 12 h light : dark environment provided by the ward, and instituting feeding by mouth, had, in most cases, only modest effects upon either circadian or ultradian rhythms. Thus, circadian rhythms continued (but generally with a period not exactly equal to 24 h), and ultradian rhythms, when present, often did not show periods that could be related easily to feeding or care-giving. These results are discussed in terms of evidence for endogenous and exogenous origins of the observed rhythms, and of theories that have postulated the relationship between circadian and ultradian rhythms. It is concluded that the results from the present analyses are difficult to reconcile with the view that circadian rhythms develop from interactions between ultradian oscillators. We suggest that they indicate a matu-ration of the circadian system as a consequence of increasing associations between the circadian elements that are present in the suprachiasmatic nuclei and in other oscillators of the circadian system. The new analytical methods used here also indicate that the results obtained from time-frequency analysis depend to some extent upon the method used.     

Karyotypes of 3- or 4-day-old pig embryos after short in-vitro culture

Pig embryos (3-4 days old) were grown for approximately 21 h in microdroplets under mineral oil with vinblastin sulphate as the mitotic inhibitor. From 10 gilts, 115 eggs were cultured: 8 proved to be unfertilized oocytes, 9 did not contain mitotic figures, 16 contained fewer than 3 good mitotic figures and 80 were judged to be of normal karyotype. A few polyploid cells were encountered, possibly of trophoblast origin. One embryo was a haploid and another one a 2N/6N mixoploid. In one gilt, in which there was no evidence of fertilization, a tetraploid embryo was discovered, and was assumed to be parthenogenetic.