Chromatin-bound PCNA as S-phase marker in mononuclear blood cells of patients with acute lymphoblastic leukaemia or multiple myeloma.

OBJECTIVES: Proliferating cell nuclear antigen (PCNA) has often been used as a marker to aid assessment of tumour growth fraction. This paper addresses the question of whether it can be used as an S-phase marker, when the non-chromatin-bound form of the protein is removed by pepsin treatment. MATERIALS AND METHODS: Cytofluorometric measurements were carried out after immunofluorescence staining of PCNA and counterstaining of DNA. S-phase fraction was determined with the help of windows on PCNA versus DNA scattergrams, or mathematically from DNA histograms. RESULTS: S-phase fractions obtained using the two methods correlated well, but did not always agree, exact discrepancies depending on the mathematical model used for histogram analysis. CONCLUSIONS: Determination of S-phase fractions with the help of PCNA immunofluorescence staining is possible, and probably more reliable than calculation of S-fractions from DNA histograms. It thus offers an alternative to assays involving BrdU labelling in vivo.

In vivo radioprotection by alpha-TMG: preliminary studies.

alpha-TMG is a novel water-soluble derivative of Vitamin E that has shown excellent antioxidant activity. The parent compound has demonstrated protection against radiation induced chromosomal damage in vivo. Hence, the preliminary experiments to determine the radioprotective activity of alpha-TMG were carried out in adult Swiss albino mice. Acute toxicity of the drug was studied taking 24h, 72 h and 30 day mortality after a single intraperitoneal injection of 500-2000 mg/kg body weight of the drug. The drug LD(50) for 24h and 72 h/30 day survival were found to be 1120 and 1000 mg/kg body weight, respectively. The optimum time of drug administration and drug dose-dependent effect on in vivo radiation protection of bone marrow chromosomes was studied in mice. Injection of 600 mg/kg of the drug 15 min before or within 5, 15 or 30min after 3Gy whole body gamma radiation resulted in a significant decrease in the aberrant metaphases percent at 24h post-irradiation; the maximum effect was seen when the drug was given immediately after irradiation. Injection of 200-800 mg/kg TMG within 5 min of irradiation with 3 Gy produced a significant dose-dependent reduction in the radiation induced percent aberrant metaphases and in the frequency of micronucleated erythrocytes at 24h after exposure, with a corresponding decrease in the different types of aberrations. The optimum dose for protection without drug toxicity was 600 mg/kg body weight. At this dose, TMG produced 70 and >60% reduction in the radiation induced percent aberrant metaphases and micronucleated erythrocytes, respectively. The high water solubility and effectiveness when administered post-irradiation favor TMG as a likely candidate for protection in case of accidental exposures.

Effect of early fetal irradiation on the postnatal development of mouse.

BACKGROUND: An earlier study has shown that irradiation at the late fetal stage of Swiss albino mice disturbed postnatal growth and appearance of physiological markers. The present study was done to determine the effect of irradiation at the early fetal stage on the postnatal development of mouse. METHODS: Fourteen-day pregnant Swiss albino mice were exposed to 0.1-1.5 Gy of cobalt-60 gamma-rays. F1 pups were observed for < or = 6 weeks of age. Postnatal mortality, body weight and length, head length and width, tail length, and the time of appearance of physiological markers (pinna detachment, eye opening, fur development, vaginal opening and testes descent) were noted. RESULTS: There was no increase in congenital anomalies. Postnatal mortality and percentage of growth-retarded pups increased significantly at doses of 0.5-1.5 Gy. A significant delay in the appearance of all the physiological markers was also noted at these doses. Body length, head length, and tail length remained significantly lower than in the controls throughout the observation period at doses of 0.3-1.5 Gy, whereas body weight and head width showed such a persistent change only at > or = 0.5 Gy. CONCLUSIONS: The early fetal day 14 in mouse is sensitive to radiation-induced postnatal mortality and impairment of growth and temporal development of physiological markers, but not to induction of congenital anomalies. While mortality and physiological markers are not affected at <0.5 Gy, growth retardation appears to have a lower threshold of approximately 0.3 Gy.

In vivo radioprotective effect of Moringa oleifera leaves.

Radioprotective property of Moringa oleifera leaves was investigated in healthy adult Swiss albino mice. Animals were injected (ip) with 150 mg/kg body weight of 50% methanolic extract (ME) of M. oleifera leaves, as a single dose, or in 5 daily fractions of 30 mg/kg each, and exposed to whole body gamma irradiation (RT, 4 Gy) 1 hr later. Five animals from each group were sacrificed at 1, 2 and 7 days after treatment. Bone marrow protection was studied by scoring aberrations in metaphase chromosomes and micronucleus induction in polychromatic erythrocytes and normochromatic erythrocytes. Pretreatment with a single dose of 150 mg/kg ME significantly reduced the percent aberrant cells to 2/3rd that of RT alone group on day 1 and brought the values to normal range by day 7 post-irradiation. A similar effect was also seen for the micronucleated cells. Fractionated administration of ME (30 mg/kg x 5) gave a higher protection than that given by the same dose administered as a single treatment. ME also inhibited the Fenton reaction-generated free radical activity in vitro in a concentration dependent manner. These results demonstrate that pretreatment with the methanolic leaf extract of M. oleifera confers significant radiation protection to the bone marrow chromosomes in mice and this may lead to the higher 30 day survival after lethal whole body irradiation.

Induction of chromosomal instability in mouse hemopoietic cells by fetal irradiation.

Mice were exposed to 0.25-1.5Gy of gamma radiation on day 14 or 17 of gestation and chromosomal aberrations were scored in the bone marrow at 12 months of age. Irradiation had resulted in low peripheral blood counts, while some animals developed very high leukocyte counts. Exposed animals showed a significant dose dependent increase in the number of aberrant metaphases, compared to unexposed animals. Fragments and polyploidy were the major types of aberrations. Mice with abnormally high blood leukocyte counts showed a higher incidence of chromosomal aberrations, especially high levels of polyploidy than in animals with low blood counts. It is concluded that radiation induced genomic instability in the fetal hemopoietic cells of mouse is transmitted to postnatal and adult bone marrow which may lead to the development of hematological disorders, including malignancies.

Influence of gestational age to low-level gamma irradiation on postnatal behavior in mice.

The present investigation was carried out to study the effects of in utero exposure to low-level gamma radiation (0.25, 0.35, or 0.50 Gy) on the postnatal neurophysiology and neurochemistry of the mouse. Pregnant Swiss albino mice were irradiated on days 11.5, 12.5, 14.5, or 17.5 post coitus (PC) and allowed to deliver. Locomotor and exploratory activities, learning and memory functions, and emotional activities were tested at 3 months of age using behavior tests. A representative group of animals was killed and hippocampal biogenic amines, noradrenaline, dopamine, serotonin (5-HT), and 5-HT's metabolite 5-hydroxy indoleactetic acid (5-HIAA), were measured. Exposure to 0.25 Gy at any of the gestation days did not produce any significant impairment in brain functions. However, an increase in gamma irradiation to 0.50 Gy on all the gestation days produced significant impairment in locomotor (open-field test) and anxiolytic (light and dark area test) activities, learning (hole board test), memory functions (active avoidance test), and emotional activity (rearings). The late fetal period is relatively resistant to radiation-induced impairment of brain functions. Both of the organogenesis gestation days showed a higher sensitivity than the fetal gestation days studied. Even a lower dose of 0.35 Gy when exposed on the late organogenesis days 11.5 and 12.5 PC, produced significant reduction in locomotor and exploratory activities. Day 11.5 PC showed a higher sensitivity than the other PC days studied. Biogenic amines did not show significant change after any of the exposures on any of the gestation days. The results suggest a threshold between 0.25 to 0.35 Gy for postnatal neurobehavior changes.

Evaluation of the cytogenetic damage and progenitor cell survival in foetal liver of mice exposed to gamma radiation during the early foetal period.

PURPOSE: To investigate the haemopoietic response to low dose gamma irradiation at the early foetal period when the liver is the major haemopoietic organ. MATERIALS AND METHODS: Pregnant Swiss albino mice were exposed to 0.1-1.5 Gy of gamma radiation on the 14th day of gestation. Twenty-four hours (15 day post conception (p.c.)) and 72 h (17 day p.c.) after exposure, the foetuses were dissected out, weighed, and liver weight and mean cellularity were determined. Cytogenetic damage in liver cells was assessed by chromosome aberration analysis and micronucleus (MN) count. The haemopoietic progenitor cell survival at 24 h and 72 h after exposure was measured by exogenous spleen colony assay on day 8 (CFU-S8) and day 12 (CFU-S12) after intravenous injection of the foetal liver cells into adult bone marrow-ablated recipient mice. RESULTS: The foetal body weight at 24 h after exposure showed a significant reduction at doses of 0.5 Gy and above, while the 72 h body weight was significantly lower than control from 0.3 Gy onwards. Liver weight showed a similar reduction for doses from 0.25 to 1.5 Gy at both the post-irradiation observation times. However, when liver weight/body weight ratios were compared, there was no significant difference between the irradiated and control values. Total liver cellularity at 24 h and 72 h after exposure showed a dose-dependent decrease, with significant depletion from control at 0.25 Gy and above. When donor cells were taken at 24 h after exposure (15 day p.c.) the CFU-S8 showed a significant decrease only at 1.0 and 1.5 Gy, while the CFU-S12 suffered such a depletion at 0.25-1.5 Gy. For donor cells recovered at 72 h after exposure, both CFU-S8 and CFU-S12 decreased linear-quadratically with radiation dose and were significantly lower than control at 0.25 Gy. A significant increase in the percent aberrant metaphases and micronucleus counts was seen at 0.1 Gy and 0.15 Gy, respectively, and increased linear-quadratically with radiation dose. CONCLUSIONS: The results demonstrate that the liver, which is the major haemopoietic organ at the early foetal period, is highly sensitive to radiation damage from maternal irradiation. At low doses, the lethal effect on the haemopoietic stem cells appears to develop more slowly than at higher doses.

Radiosensitization of a mouse melanoma by withaferin A: in vivo studies.

The radiosensitizing effect of a plant withanolide, withaferin A, on the B16F1 mouse melanoma was studied in vivo. Treatment of 100 mm3 tumours with 10 to 60 mg/kg withaferin A intraperitoneally produced a dose dependent increase in growth delay and volume doubling time. Injection of 30-50 mg/kg withaferin A, followed by 30 Gy local gamma irradiation, significantly enhanced the tumour response. No systemic or local adverse reactions were noted in these groups. The drug was most effective when injected intraperitoneally 1 h before irradiation. However, neither the individual agents nor their combination could produce any complete response (tumour cure). Melanoma is a relatively radioresistant tumour. The present results indicate that the radiation response of this tumour can be significantly enhanced by pretreatment with withaferin A.

Modification of radiation induced damage in mouse intestine by WR-2721.

Intestinal protection in mice against radiation injury by WR-2721 (300 mg/kg body wt, i.p., 30 min before irradiation) was studied after whole body gamma irradiation (0.5, 1.5, 3.0, 4.5, or 6.0 Gy). Crypt survival and induction of apoptosis, and abnormal mitoses in crypt cells in the jejunum were studied on day 1, 3 and 7 after irradiation. Irradiation produced a significant decrease in crypt survival, whereas apoptosis and abnormal mitoses showed a significant increase from sham-treated control animals. Maximum changes in all the parameters were observed on day 1 after irradiation and the effect increased linearly with radiation dose. There was recovery at later intervals, which was inversely related to radiation dose. WR-2721 pre-treatment resulted in a significant increase in the number of surviving crypts, whereas the number of apoptotic cells in the crypts showed a significant decrease from respective irradiated controls on day 1 after exposure. The recovery was also faster in WR-2721 pre- treated animals. It is concluded that WR-2721 protects against gastrointestinal death by reducing radiation induced cell death, thereby maintaining a higher number of stem cells in the proliferating compartment.

Evaluation of anti-tumor efficacy of injectable Centchroman in mice bearing Ehrlich ascites carcinoma.

Anti-tumor efficacy of Centchroman formulated as niosomes and gel implant was evaluated in Swiss albino mice bearing Ehrlich ascites carcinoma at 10 mg/kg body weight dose given subcutaneously. Median day of death, percentage increase in host life span and changes in body weight were studied. Centchroman significantly (P < 0.05) increased the median day of death both in free and formulated systems. Also, injectable formulations exhibited a significant (P < 0.05) increase in host life span compared to free drug, hence, enhanced anti-tumor efficacy against Ehrlich ascites carcinoma.

Effect of prenatal gamma irradiation during the late fetal period on the postnatal development of the mouse.

Pregnant Swiss albino mice were exposed to 0.3, 0.5, 1.0, or 1.5 Gy of gamma radiation on day 17 of gestation. Sham-exposed controls were examined for comparison. Exposed mice as well as controls were left to complete gestation and parturition. Pups were observed up to age 6 weeks; appearance of physiological markers (pinna detachment, eye opening, fur development, vaginal opening, and testes descent), postnatal mortality, body weight, body length, head length, head width, and tail length were recorded. A significant delay in fur development was observed at 0.3 Gy and in other physiological markers at doses above 0.3 Gy, while a significant increase in mortality and growth retardation occurred only at 1.0 and 1.5 Gy. Although congenital anomalies such as syndactyly and bent tail were observed at doses of 0.5-1.5 Gy, only syndactyly showed a statistically significant increase in frequency. A statistically significant lower body weight was observed during the first week of postnatal life, but body weights increased to normal levels by the second week in animals exposed to doses less than 1.0 Gy. At higher doses, low body weight persisted throughout the postnatal period. Head length and tail length showed a significant decrease from controls at 0.5-1.5 Gy, and the effect was evident from birth to age 6 weeks. But a similar effect on body length and head width was noticed only at 1.0 and 1.5 Gy. These studies indicate that even in the absence of any major morphological changes, normal development of physiological landmarks and postnatal growth can be impaired by fetal irradiation at 17 days p.c. (post coitus). Morphological changes appear to have a threshold between 0.3-0.5 Gy, while physiological marker effects may occur with a lower threshold.

Quantitative estimation and recommendations for supplementation of protein lost through scaling in exfoliative dermatitis.

BACKGROUND: Exfoliative dermatitis (ED) can result in protein loss due to scaling causing a negative nitrogen balance. Freedberg and Baden (J Invest Dermatol 1962; 38: 277-284) estimated the amount of scale lost in ED by collecting it in an occlusive suit. Subsequently, the nitrogen content was determined by the Kjeldahl method. The exact amount of protein supplementation in ED, dependent on scale loss, is not well established. As occlusion and hyperthermia caused by the suit can inhibit scaling, the objectives of the present study were to design an alternative method to measure the amount of scale lost, to estimate the protein content of the scale, and to propose suitable recommendations for protein supplementation. METHODS: In 40 patients with ED, the total protein content lost through scaling per day (P) was determined by the following equation: P = TxIxYxX/25x10(4) g, where T is the total body surface area in square meters, I is the percentage area involved in scaling, estimated using computer-aided design (CAD graph), Y is the amount of scale lost per unit area (0.0025 m2) in milligrams, and X is the quantity of protein present in 1 g of scale in milligrams estimated by a spectrophotometer. RESULTS: It was observed that patients with ED secondary to drug reactions, eczema, and psoriasis lost 7.2, 9.6, and 22.6 g of scale with a protein content of 4.2, 5.6, and 12.8 g respectively. The difference in the amount of protein lost in ED secondary to drug reactions and eczema was not statistically significant; however, the protein lost in psoriasis was significant (p < 0.01 to p < 0.05). CONCLUSIONS: ED may increase the daily protein loss by approximately 25-30% in psoriasis and 10-15% in other causes. Standard treatment for ED and protein supplementation based on our recommendations can minimize the adverse effects of a negative nitrogen balance.

Effect of late fetal irradiation on adult behavior of mouse: Dose-response relationship.

Pregnant Swiss mice were exposed to 0.3-1.5 Gy of gamma radiation on day 17 of gestation and allowed to deliver the offspring. When the F1 mice were 6 months old, they were subjected to a number of behavioral tests. Open-field and dark-bright arena tests were conducted to study locomotor and exploratory activities. Learning and memory were tested by holeboard activity, conditioned avoidance response, and radial arm maze performance. After all the tests, 20 animals (10 males and 10 females) from each group were killed, and their brain weight was taken. The open-field and dark-bright arena tests showed a significant dose-dependent decrease in the locomotor and exploratory activities. Reduction in time spent in the dark area and higher locomotor activity in the bright area indicated a reduced aversion to bright light. But the emotional activities like rearing and grooming did not change. The learning and memory functions also showed a significant impairment, even at 0.3 Gy. The deficit in the performance in the holeboard test, conditioned avoidance response, as well as maze-learning efficiency, decreased linearly with increase in radiation dose. The brain weight showed a linear dose-dependent decrease. But the brain/body weight ratio was not significantly affected even at 1.5 Gy. These results demonstrate that exposure of a mouse on day 17 of gestation to radiation doses below 1.0 Gy can induce significant impairment in the adult brain function, without producing any notable effects on brain morphology. This study also suggests that the retardation of higher brain function by exposures during the late fetal period may have a threshold of around 0.3 Gy.

Modulation of glutathione and antioxidant enzymes by Ocimum sanctum and its role in protection against radiation injury.

Aqueous extract (OE) of the leaves of Ocimum sanctum, the Indian holy basil, has been found to protect mouse against radiation lethality and chromosome damage and to possess significant antioxidant activity in vitro. Therefore a study was conducted to see if OE protects against radiation induced lipid peroxidation in liver and to determine the role, if any, of the inherent antioxidant system in radioprotection by OE. Adult Swiss mice were injected intraperitoneally (i.p.) with 10 mg/kg of OE for 5 consecutive days and exposed to 4.5 Gy of gamma radiation 30 min after the last injection. Glutathione (GSH) and the antioxidant enzymes glutathione transferase (GST), reductase (GSRx), peroxidase (GSPx) and superoxide dismutase (SOD), as well as lipid peroxide (LPx) activity were estimated in the liver at 15 min, 30 min, 1, 2, 4 and 8 hr post-treatment. LPx was also studied after treatment with a single dose of 50 mg/kg of OE with/without irradiation. OE itself increased the GSH and enzymes significantly above normal levels whereas radiation significantly reduced all the values. The maximum decline was at 30-60 min for GSH and related enzymes and at 2 hr for SOD. Pretreatment with the extract checked the radiation induced depletion of GSH and all the enzymes and maintained their levels within or above the control range. Radiation significantly increased the lipid peroxidation rate, reaching a maximum value at 2 hr after exposure (approximately 3.5 times that of control). OE pretreatment significantly (P < 0.0001) reduced the lipid peroxidation and accelerated recovery to normal levels. The results indicate that Ocimum extract protects against radiation induced lipid peroxidation and that GSH and the antioxidant enzymes appear to have an important role in the protection.

Effect of plumbagin on the radiation induced cytogenetic and cell cycle changes in mouse Ehrlich ascites carcinoma in vivo.

The effect of plumbagin, a naphthoquinone from the roots of the Indian medicinal plant Plumbago rosea, and Cobalt-60 gamma radiation was studied on Ehrlich ascites carcinoma in vivo, taking cytogenetic damage and cell cycle changes as experimental endpoints. Plumbagin (5 mg/kg body wt, P1) administered intraperitoneally produced a significant increase in the percentage of S-phase as well as G2-M cells with a corresponding decrease in the G1 phase at different post-treatment times. Radiation (7.5 Gy, RT) alone produced the classical G2 block at 1 hr, which persisted with a continuous increase throughout the post-treatment observation period. The combination treatment produced a similar effect as that of RT on G2-M cells, but its effect on the G1 phase was more pronounced than the latter. While P1 treatment produced a small increase in the percentage of labeled S-phase cells, combination treatment significantly reduced the labeled S-phase cells with a corresponding increase in the unlabeled fraction. Drug or radiation alone significantly increased micronuclei induction at various post-treatment times and the combination of the two further enhanced this effect additively. The mechanism of interaction of P1 with radiation in bringing about this effect is not clear.

Cytotoxic and anticancer activities of some 1-aryl-2-dimethylaminomethyl-2-propen-1-one hydrochlorides.

A series of 1-aryl-2-dimethylaminomethyl-2-propen-1-one hydrochlorides demonstrated marked cytotoxicity towards approximately 55 human tumour cell lines from different neoplastic diseases. In general they were more potent than melphalan and displayed selective toxicity towards human leukemic cells. A representative compound, 1-phenyl-2-dimethyl-aminomethyl-2-propen-1-one hydrochloride (2a), had similar cytotoxicity as melphalan towards murine P388 and L1210 leukemic cells. In addition, 2a reduced the sizes of a number of human tumour xenografts including colon, prostatic and melanotic cancers passaged in athymic mice. Compound 2a showed excellent activity towards Ehrlich ascites carcinoma and B16F1 melanoma in mice which was enhanced using niosomes. One may conclude from the data generated that 1-aryl-2-dimethylaminomethyl-2-propen-1-one hydrochlorides are a novel series of cytotoxic and anticancer agents.

A comparative study of radioprotection by Ocimum flavonoids and synthetic aminothiol protectors in the mouse.

The radioprotective effects of two flavonoids, orientin (Ot) and vicenin (Vc), obtained from the leaves of Ocimum sanctum, and the synthetic compounds WR-2721 and MPG (2-mercaptopropionyl glycine) have been compared by examining chromosome aberration in cells of bone marrow in irradiated mice. Healthy adult Swiss mice were injected intraperitoneally (i.p.) with 50 micrograms kg-1 body weight of Ot or Vc; 20 mg kg-1 of MPG; 150 mg kg-1 of WR-2721 or double distilled water (DDW). They were exposed to whole body irradiation of 2.0 Gy gamma radiation 30 min later. After 24 h, chromosomal aberrations were studied in the bone marrow of the femur by routine metaphase preparation after colchicine treatment. Radiation (2 Gy) increased the number of aberrant cells from less than 1% in controls to almost 20%. Pre-treatment with all the protective compounds resulted in a significant reduction in the percentage of aberrant metaphases as well as in the different types of aberration scored. Vc produced the maximum reduction in percent aberrant cells while MPG was the least effective; Ot and WR-2721 showed an almost similar effect. However, WR-2721 was the most effective against reduction of complex an almost similar effect. However, WR-2721 was the most effective against reduction of complex aberrations, followed by Vc. Neither flavonoids had any systemic toxicity, even at 200 mg kg-1 body weight. Considering the low dose needed for protection and the high margin between the effective and toxic doses, the ocimum flavonoids may be promising for human radiation protection.

Enhancement of bone marrow radioprotection and reduction of WR-2721 toxicity by Ocimum sanctum.

The radioprotective effect of the leaf extract of Ocimum sanctum (OE) in combination with WR-2721 (WR) was investigated on mouse bone marrow. Adult Swiss mice were injected intraperitoneally (i.p.) with OE (10 mg/kg on 5 consecutive days), or 100-400 mg/kg WR (single dose) or combination of the two or double-distilled water (DDW) and whole-body exposed to 4.5 Gy gamma-irradiation (RT). Metaphase plates were prepared from femur bone marrow on days 1, 2, 7 and 14 post-treatment and chromosomal aberrations were scored. The maximum number of aberrant cells was observed at 24 h after irradiation in all the groups. However, pretreatment with OE or WR individually resulted in a significant decrease in aberrant cells as well as different types of aberrations. The combination of the two further enhanced this effect; resulting in a 2-fold increase in the protection factor (PF = 6.68) compared to 400 mg/kg WR alone. The percent aberrant cells decreased linear-quadratically with WR dose when given individually, while in the OE + WR pretreatment animals the values showed a linear dose response. Combination of OE with WR doses above 200 mg/kg completely eliminated rings, polyploidy and pulverization of chromosomes. Percent aberrant cells decreased with time in all groups, though the values remained higher than normal even on day 14 in the RT alone as well as those treated with single agent + RT. WR doses above 200 mg/kg before RT resulted in significantly higher frequency of aberrant cells compared to RT and OE + RT groups on day 14, suggesting delayed WR toxicity; but combination of OE with WR brought down these values to normal level, indicating that OE combination, in addition to enhancing WR protection, may also act as a detoxifier. The protective effect of OE and WR is also reflected in the enhancement of bone marrow CFU survival. Both OE and WR possessed significant free radical scavenging activity in vitro. The combination of the two further enhanced this effect, suggesting that the enhanced free radical scavenging activity by combining the two protectors results in the higher bone marrow cell protection. The significant elevation in chromosome protection obtained by combining OE with WR, with reduction in the latter's toxicity at higher doses, suggests that the combination may have promise for radioprotection in humans.

Modification of bone marrow radiosensensitivity by medicinal plant extracts.

Withaferin A (WA), a steroidal lactone, and Plumbagin (Pl), a naphthoquinone, from the roots of Withania somnifera and Plumbago rosea, respectively, have been shown to possess growth inhibitory and radiosensitizing effects on experimental mouse tumours. An aqueous extract of the leaves of Ocimum sanctum (OE) was found to protect mice against radiation lethality. Therefore, the radiomodifying effects of the above plant products on the bone marrow of the adult Swiss mouse was studied. Single doses of WA (30 mg kg-1) or Pl (5 mg kg-1) were injected intraperitoneally (ip) and OE (10 mg kg-1) was injected ip once daily for five consecutive days. Administration of extracts was followed by 2 Gy whole body gamma irradiation. Bone marrow stem cell survival was studied by an exogenous spleen colony unit (CFU-S) assay. The effects of WA and Pl were compared with that of cyclophosphamide (CP) and radioprotection by OE was compared with that of WR-2721 (WR). Radiation reduced the CFU-S to less than 50% of normal. WA, CP and Pl significantly enhanced this effect and reduced the CFU-S to almost the same extent (to < 20% of normal), although individually WA and Pl were less cytotoxic than CP. These results indicate that radiosensitization by WA and Pl is not tumour specific. OE significantly increased CFU-S compared with radiotherapy (RT) alone. OE+RT gave a higher stem cell survival (p < 0.05) than that produced by WR+RT. While WR alone had a toxic effect, OE treatment showed no such effect, suggesting that the latter may have an advantage over WR in clinical application.

Protection against radiation-induced chromosome damage in mouse bone marrow by Ocimum sanctum.

The radioprotective effect of the leaf extract of Ocimum sanctum (Ocimum extract, OE) was investigated by taking chromosome aberrations as the end point. Adult Swiss mice were whole-body exposed to 1-6 Gy of gamma radiation with/without pretreatment with 10 mg/kg b.wt. of OE intraperitoneally for 5 consecutive days. Radiation was given 30 min after the last injection. Metaphase plates were prepared from femur marrow on days 1, 2, 7 and 14 post-treatment and the frequency of aberrant cells and individual aberrations were scored. OE alone did not have any significant effect on the chromosomes. Maximum percent of aberrant cells was observed at 24 h in all the exposed groups. The percent aberrant cells showed a linear quadratic increase with radiation dose, in both radiation alone (RT) and OE + RT-treated groups. Exchange (dicentrics and rings) and multiple (pulverized and severely damaged cells) aberrations also showed a similar response. However, the slopes of OE + RT was significantly shallower than RT groups (p < 0.05). A dose-modifying factor of 2.63 was obtained taking percent aberrant cells for 2 Gy as the base. Progressive decline in the percent aberrant cells as well as the number of aberrations with time after irradiation was observed in both RT and OE + RT groups. OE treatment resulted in a faster recovery compared to RT alone group. At doses below 3 Gy, OE pretreatment almost completely eliminated the exchange aberrations from the cell population by day 2. Studies on a chemical system demonstrated that OE significantly reduced the generation of hydroxyl radical; a lower dose of OE (1 mg/ml) was more effective than 5 mg/ml and this effect was more pronounced than that produced by DMSO. These results show that OE affords in vivo protection against radiation-induced cytogenetic damage. Free radical scavenging is a likely mechanism of OE protection.