Effects of blue pulsed light on human physiological functions and subjective evaluation

Background

It has been assumed that light with a higher irradiance of pulsed blue light has a much greater influence than that of light with a lower irradiance of steady blue light, although they have the same multiplication value of irradiance and duration. We examined the non-visual physiological effects of blue pulsed light, and determined whether it is sensed visually as being blue.

Findings

Seven young male volunteers participated in the study. We placed a circular screen (diameter 500 mm) in front of the participants and irradiated it using blue and/or white light-emitting diodes (LEDs), and we used halogen lamps as a standard illuminant. We applied three steady light conditions of white LED (F0), blue LED + white LED (F10), and blue LED (F100), and a blue pulsed light condition of a 100-μs pulse width with a 10% duty ratio (P10). The irradiance of all four conditions at the participant''s eye level was almost the same, at around 12 μW/cm². We measured their pupil diameter, recorded electroencephalogram readings and Kwansei Gakuin Sleepiness Scale score, and collected subjective evaluations. The subjective bluish score under the F100 condition was significantly higher than those under other conditions. Even under the P10 condition with a 10% duty ratio of blue pulsed light and the F10 condition, the participant did not perceive the light as bluish. Pupillary light response under the P10 pulsed light condition was significantly greater than under the F10 condition, even though the two conditions had equal blue light components.

Conclusions

The pupil constricted under the blue pulsed light condition, indicating a non-visual effect of the lighting, even though the participants did not perceive the light as bluish.     

Analysis of chromosomes from human peripheral lymphocytes by flow cytometry

A method of preparation and flow cytometric analysis of chromosomes from human peripheral lymphocytes is described. The procedure allows a resolution coefficient of variation better than 3% using propidium iodide staining and a commercially available flow cytometer.     

Studies on the chromosomes of human lymphocytes treated with diazepam in vitro

Human lymphocytes were exposed in vitro to various concentrations of the psychotherapeutic agent diazepam, using two different techniques for culturing the lymphocytes, a whole-blood and a macro procedure. There was no evidence for a damaging effect of diazepam on the lymphoblast chromosomes at any concentration or exposure time studied with either technique of culture. The significance of results obtained in vitro on chromosome-breaking effects of chemical agents in commercial use is discussed, and the importance of some technical details in conducting such experiments is pointed out.     

Effects of chlorambucil on human chromosomes

No significant amount of chromosomal damage was found in the 48-h cultures of lymphocytes of 18 patients who had been treated with the bifunctional alkylating agent chlorambucil (CBC). However, there was suggestive evidence of chromatid damage (i.e. of types attributable to damage during or after DNA synthesis in the cell cycle). In marrow cells of 3 patients given a single large dose of chlorambucil (equivalent to 2 days' normal treatment) there was also suggestive evidence of induced chromatide-type damage.Extensive series of in vitro experiments yielded evidence that (a) exposure of human lymphocytes over the whole period of culture showed chromatid-type damage; (b) this damage increased sharply from concentrations of 0.5 μg/ml to3.0 μg/ml; (c) although chromatide-type damage always predominated, there was suggestive evidence also of chromosome-type aberrations attributable to damage occuring in the G₀/G₁ period, although some or all of this could be attributed to “derived” chromatid damage; (d) even if lymphocytes were only exposed during the G₀ or G₁ periods of the cycle, damage was found in the subsequent metaphases and it was almost entirely of the chromatid type; (e) much more damage occurred in lymphocytes exposed for varying periods to the drugs after stimulation by phytohaemagglutinins than in those exposed in whole blood, or in medium before stimulation; (f) damaged occurred in lymphocytes exposed to the drug while in S but not exposed only when in G₂; (g) no evidence was found that unschaduled DNA synthesis during G₀ or G₁ was induced by the drug; (h) there appeared to be no delay caused by the drug in the time at which cells reached the first “S” phase in culture but there was some evidence consistent with prolongation of “S” in cells exposed in culture; (i) there was evidence that CBC alone could stimulate lymphocyte tto DNA synthesis, and that a few cells proceeded in the cycle to prophase, or even metaphase. However, there was a considerable amount of cell-killing during CBC-stimulated DNA synthesis.     

Effects of LSD on human chromosomes

A number of positive and negative studies have been reported with regard to the damaging effects of LSD on human chromosomes. The present report describes a comparative study of cytogenetic analyses of 200 metaphases of lymphocytes from each of 6 subjects (3 males, 3 females) at varying concentrations of LSD, along with a positive control with mitomycin C and a negative control with sterile water. Results of a small pilot study on the effects of LSD on growth, macromolecular synthesis, mutation, and recombination in bacteria, λ phage and mammalian cells are also included. The data failed to show any significant differences between chromosome aberrations and LSD. Significant changes in somatic cells and in chromosomes occurred only at high doses of LSD.     

Effects of CCNU therapy on human chromosomes

Peripheral blood lymphocytes of 9 patients under CCNU therapy were examined for frequency of sister-chromatid exchanges (SCEs) and chromosomal aberrations (CAs). 7 out of 9 patients were treated with only CCNU, whereas the remaining 2 were treated with other chemotherapeutic agents in combination with CCNU. Compared to normal individuals, a significantly increased frequency of SCE was observed in the patients before starting anticancer therapy (P less than 0.001). Increased incidences of structural changes in chromosomes were observed in cells from all the treated patients. The most frequent aberrations were of chromatid type. After administration of a single dose of CCNU, an increase in SCE frequencies was observed which remained elevated even after 6 weeks. It was concluded that increases in SCEs and CAs in lymphocytes were caused by CCNU treatment. Further studies are needed to elucidate whether any CAs observed in the present study could participate in the induction of second neoplasm.     

The effects of busulphan on the chromosomes of normal human lymphocytes

In vitro exposure of human lymphocytes to busulphan (BUS) produced an increase in chromosome aberrations and in sister-chromatid exchange (SCE) frequency. The distribution of chromosome breaks throughout the karyotype was non-random and they occurred mainly in the G-negative bands. Certain bands had a marked susceptibility to BUS and comparisons with the human chromosome-break distributions reported for a number of drugs revealed that some of these bands were equally susceptible to other alkylating agents. Both the number of chromosome gaps and breaks and the SCE frequency increased with BUS concentration, but only the SCE dose-response was a clearly defined linear relationship. Therefore a standard SCE dose-response curve was constructed for future comparison with the results of similar investigations of patients on BUS therapy.     

The effects of busulphan on the chromosomes of normal human lymphocytes

In vitro exposure of human lymphocytes to busulphan (BUS) produced an increase in chromosome aberrations and in sister-chromatid exchange (SCE) frequency. The distribution of chromosome breaks throughout the karyotype was non-random and they occurred mainly in the G-negative bands. Certain bands had a marked susceptibility to BUS and comparisons with the human chromosome-break distributions reported for a number of drugs revealed that some of these bands were equally susceptible to other alkylating agents. Both the number of chromosome gaps and breaks and the SCE frequency increased with BUS concentration, but only the SCE dose--response was a clearly defined linear relationship. Therefore a standard SCE dose--response curve was constructed for future comparison with the results of similar investigations of patients on BUS therapy.     

Effects of gamma-irradiation on chromosomes of cultured lymphocytes from rheumatoid arthritis patients and their family members

Increased rates of spontaneous or induced chromosome breakage are seen in many types of diseases, including the 'collagen-type' autoimmune diseases. Using a 60Co gamma cell, we irradiated lymphocyte cultures from three related rheumatoid arthritis patients, their immediate family members, and two unrelated rheumatoid arthritis patients. Although these individuals had not shown abnormally high levels of spontaneous chromosome breakage, they did show an abnormal sensitivity to irradiation, which was manifested in several ways. Two of the probands showed induced breakage rates that were twice as high as those seen in controls. In addition, the reduction of mitotic index, due either to increased cell death or to induction of a G2 lag period, was higher in the arthritis group (including non-symptomatic family members) than in the control group. Finally, we observed a high frequency of an unusual type of cell in the arthritis group. These unusual cells resembled c-anaphases seen with extended colcemid treatment, and may indicate that the mitotic apparatus in cells from this group is particularly sensitive to ionizing radiation.     

Banding of unfixed mitotic chromosomes in suspension after release from human lymphocytes and fibroblasts

Summary Combined application during chromosome isolation of the non- or weakly fluorescent DNA-intercalators 4-aminomethyl-4,5, 8-trimethylpsoralen and daunomycin as stabilizers of mitotic chromosome structure, and the non-intercalating DNA-binding fluorochromes DAPI and D287/170 as producers of a visible banding pattern, resulted in clearly banded unfixed floating chromosomes. Chromosomes stabilized by intercalation appeared to be sufficiently stable to allow the reproduction of distamycin A/DAPI or netropsin/DAPI staining in suspension, thus highlighting specific heterochromatic regions on the floating chromosomes. The results of this study demonstrate that the inducibility of bands is an inherent characteristic of mitotic chromosome organization. Possible practical applications of these results in flow cytometry are discussed.     

Effects of age on segregation of the X and Y chromosomes in cultured lymphocytes from Chinese men

Chromosome malsegregation in binucleated lymphocytes is a useful endpoint to evaluate age effect on genetic stability. However, the investigations on chromosome malsegregation in binucleated lymphocytes from Chinese are scarce. In this study, peripheral blood lym- phocytes were collected from 14 old （60-70 years） and 10 young （22-26 years） healthy Chinese men. To detect malsegregation of the sex chromosomes, multi-color fluorescence in situ hybridization （FISH） was performed on binucleated lymphocytes, cytokinesis-blocked by cytochalasin B at the first mitosis after phytohaemagglutinin stimulation. Compared with that in young men, a significant increase in frequencies of loss of chromosome X （9.2± 3.2‰ vs. 1.1 ± 0.9‰, P 〈 0.001） and Y （2.5 ± 1.9‰ vs. 0.2± 0.3‰, P 〈 0.001） was found in old men. Similarly, nondisjunction of chromosome X （16.5± 3.4‰ vs. 3.5 ± 1.1‰, P 〈 0.001） and Y （7.2 ± 2.6‰ vs. 2.4 ± 1.3‰, P 〈 0.001） occurred more frequently in old men than in young men. Regardless of donor＇s age, nondisjunction is more prevalent than loss for both chromosome X and Y. The frequencies of observed simultaneous malsegregation were relatively higher than the expected, suggest- ing an association between malsegregation. These results indicated that in Chinese men, malsegregation of the sex chromosomes increases with age in an associated fashion, and nondisjunction accounts for the majority of spontaneous chromosome malsegregation.     

A technique for simultaneous antikinetochore immunofluorescence staining and Q-banding in chromosomes from human lymphocytes

Antikinetochore immunofluorescence staining has been used in several studies to determine whether a second kinetochore is present, active, or both, in multicentric chromosomes. All of these studies have used tissue culture cells, and contended with the problem of obtaining well spread, banded metaphase chromosomes without affecting the kinetochore staining. We have adapted hypotonic, centrifugation and chromosome staining techniques to obtain simultaneous Q-banding and bright kinetochore staining of chromosomes from human lymphocytes.     

Frequency and non random distribution of silver-staining NORs on acrocentric chromosomes from neonate and adult human lymphocytes

The NORs frequency in a group of newborns and adults was determined by the gelatine silver staining technique. A higher number of Ag-NORs (χ² test, p<0.01) was found in adults than in newborns. The lack of correlation between cell proliferating rate index (PRI) and frequency of Ag-NORs let us suppose that the decrease of Ag-positive NORs in neonates could probably be due to factors different from cell kinetics. A non random distribution of Ag-NORs on the acrocentric chromosomes was also demonstrated: chromosome 21, in particular, showed the highest frequency, while chromosome 15, the lowest.     

Radiosensitivity of chromosomes in lymphocytes from Down's syndrome patients

A study was made of the yield of chromosome aberrations in gamma-irradiated G0 peripheral blood lymphocytes from 6 patients with different forms of Down's syndrome. The doses used were from 0.25 to 3.0 Gy. Seven healthy donors of different age made the control group. There was a significant increase in the yield of chromosome exchanges in lymphocytes from all the patients as compared to control. The spontaneous level of chromosome aberrations and the frequency of radiation-induced fragments did not differ from the control values. The yield of exchanges in diploid and trisomic cells from patients with the mosaic form of Down's syndrome did not change significantly as the time of cultivation was raised. The origin of DNA repair defects leading to the increased chromosome radiosensitivity in Down's syndrome is discussed.     

Effect of chromosomal constitution with respect to the sex chromosomes on the proliferative activity of human lymphocytes

Rate division of human lymphocytes was studied in 85 healthy siblings and 142 normal non-related individuals using sister chromatid differential staining in standard conditions of cell culturing. It was shown that proliferation of male lymphocytes exceeds that of female cells. This sexual dimorphism does not depend on the time of fixation of cell cultures and probably is conditioned by differential chromosomal constitution of the two sexes. The study of sex chromosome mosaics revealed rate modification of cell proliferation in the line: 45,X greater than 46,XY greater than 46,XX. The possibility of influence of heterochromatin and sex chromosomal genes on control of cell division is discussed.     

Frequency and distribution of associations of acrocentric chromosomes in human lymphocytes

Frequency and character of the distribution of acrocentric chromosome associations are determined in 40 phenotypically healthy native inhabitants of the Latvian SSR (20 males and 20 females). The ability to associations is the lowest for chromosomes 15 and 22 and the highest for chromosomes 21, 14 and 13. It is found that a tendency to associations between chromosomes 21-21 (P less than 0.05) and 13-21 (P less than 0.01) is not of an accidental character.     

Genotoxic effects of white fluorescent light on human lymphocytes in vitro

Sources of light beams such as white fluorescent light, are present in our daily life to meet the needs of life in the modern world. This study was conducted with the objective of determining the possible genotoxic, cytotoxic and aneugenic effects caused by this agent in different stages of the cell cycle (G0/early G1, S, and late G2), using different cytogenetic parameters (sister chromatid exchanges--SCE, chromosome aberrations--CA, and detection of aneugenic effects) in lymphocytes from temporary cultures of human peripheral blood. WFL showed a genotoxic effect in vitro, expressed by an increase in the frequency of SCE's, regardless of the cell cycle stage. However, no increase in the frequency of CAs was observed. In addition, disturbances in cell cycle kinetics and chromosomal segregation were also observed. Taken together, such data may contribute to a better understanding and a different management in the use of phototherapy for some pathological conditions.