Present investigation was planned to evaluate the nootropic effect of Centella asiatica. Three months old male Swiss albino mice were injected orally with graded doses (200, 500, 700, 1000 mg/kg body weight) of C. asiatica aqueous extract for 15 days to select an effective dose for nootropic studies. Animals were tested in radial arm maze to assess the learning and memory performance. Based on these results, mice were treated orally with 200 mg/kg of C. asiatica for 15 days from day 15 to day 30 post partum (p.p.) and the nootropic effect was evaluated on the 31st day and 6 months p.p. The behavioral (open field, dark/bright arena, hole board and radial arm maze tests), biochemical (acetylcholine esterase activity) and histological studies (dendritic arborization) were carried out. Performance of juvenile and young adult mice was significantly improved in radial arm maze and hole board tests, but locomotor activity did not show any change compared to control. Treatment resulted in increased acetylcholine esterase activity in the hippocampus. Dendritic arborization of hippocampal CA3 neurons was also increased in terms of intersections and branching points, both at one month and 6 months. Results of the present investigation show that treatment during postnatal developmental stage with C. asiatica extract can influence the neuronal morphology and promote the higher brain function of juvenile and young adult mice.

Purpose: To study the protective effect of orientin and vicenin against early genomic effects of foetal irradiation and their late consequences in mice. Materials and methods: Fourteen‐day pregnant mice were exposed to 1 Gy 60Co gamma‐radiation 30 min after an intraperitoneal injection of orientin or vicenin (50 µg kg−1 body weight). Chromosomal aberrations were studied in foetal liver cells and their spleen colonies (three passages, colony‐forming units‐spleen CFU‐S1, CFU‐S2, CFU‐S3) and 1–12 months post‐partum bone marrow. Peripheral blood counts and solid tumours were recorded to 12 and 20 months, respectively. Results: Irradiation significantly increased the percent aberrant cells and aberrations/cell in foetal liver and CFU‐S1. These effects decreased in later passages of CFU‐S and were not seen at 1–6 months post‐partum, but increased significantly from 9 months. Total blood counts showed significant reduction from 6 months, while neutrophils increased from 3 months post‐partum. Solid tumour incidence in adults increased significantly, with a decrease in age at detection. Orientin/vicenin significantly reduced the chromosomal anomalies in foetal and adult haemopoietic cells, restored blood counts to the normal range, and significantly reduced tumour incidence and delayed tumour development to control age. Conclusions: Orientin and vicenin protect against foetal irradiation‐induced genomic damage and instability, thereby reducing the delayed chromosomal abnormalities and tumorigenesis in adult.

A Study was carried out to see if acute low dose fetal irradiation Of Mouse could induce long lasting chromosomal anomalies and adult cancers. The abdomen of pregnant Swiss juice was exposed to 0.1-1 Gy of Co-60 gamma radiation on day 14 of gestation. The animals were killed 24 h later or allowed to deliver their young. Hemopoietic cell death and chromosomal aberrations were studied in the fetal liver. Chromosomal instability was traced in the 1 Gy exposed group from the fetal liver cells through their CFU-S to postnatal bone marrow. The F1 mice were observed for chromosomal aberrations in the bone marrow and for solid tumor incidence. Irradiation produced a significant dose-dependent increase in the hemopoietic cell death and aberrant metaphases in the fetal liver. The main types of aberrations were chromatid breaks and fragments. The aberrations persisted in the spleen colonies developed from these cells. Their frequency decreased in the postnatal bone marrow, however, showed a significant increase at 12 months of age. Some exposed animals had abnormally high leukocyte counts and after 1 Gy their bone marrow had eight times more polyploid cells than the control. Prenatal exposure also significantly increased the incidence of solid tumors, the ovaries showing the highest risk. It is concluded that a single exposure below 1 Gy of gamma radiation at the early fetal stage of these mice can induce persistent chromosomal instability in the hemopoietic cells, and significantly increase the incidence of solid tumors in adults. (C) 2002 Elsevier Science B.V. All rights reserved.

Pregnant Swiss albino mice were exposed to diagnostic levels of ultrasound (3.5 MHz, intensity 65 mW, I(SPTP) = 1 W/cm(2), I(SATA) = 240 W/cm(2)) for 10, 20 and 30 minutes on day 14 of gestation. Sham exposed controls were maintained for comparison. Fifteen pregnant mice were exposed for each group. Exposed as well as control animals were left to complete gestation and parturition. Ultrasound induced changes in maternal vaginal temperature was recorded. The changes in the physiological reflexes and postnatal mortality up to 6 weeks of age were recorded. The litters were subjected to behavioral tests for locomotor activity, learning and memory at 4 month and 1 year of age. Neither the physiological reflexes nor the postnatal mortality was affected by ultrasound exposure. However, there was a noticeable impairment in both locomotor and learning behavior even after a 10 min exposure, which further increased with increases in exposure time. Thus the present study demonstrates the neurotoxicity of diagnostic ultrasound and the high susceptibility of early fetal brain to induction of lasting detrimental changes by ultrasound exposure.

Radiotherapy is routinely employed in the treatment of head and neck cancers. Acute cell death, radiation-activated chemical cascades, and the induction of genes coding for protective factors like cytokines are considered to be the major processes involved in radiation damage and repair. It should be possible to follow these processes by monitoring the biochemical interactions initiated by radiation. We have carried out Raman spectroscopy studies on tissue from mice subjected to brain irradiation to identify the biochemical changes occurring in tissue and brain as a result of radiation injury. These studies show that brain irradiation produces drastic spectral changes even in tissue far removed from the irradiation site. The changes are very similar to those produced by the stress of inoculation and restraint and the administration of an anesthetic drug. While the changes produced by stress or anesthetics last for only a short time (a few hours to 1 or 2 days), radiation-induced changes persist even after 1 week. The spectral changes can be interpreted in terms of the observation of new spectra that are dominated by bands due to proteins. The results thus support the hypothesis that various protective factors are released throughout the body when the central nervous system (CNS) is exposed to radiation.

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.

The abdominal region of pregnant Swiss albino mice was exposed to single dose of 0.5 Gy gamma-radiation at gestation days from 1.5 to 17.5 days post-coitus (p.c). The animals were sacrificed on day 18 p.c. and foetuses were examined for resorption and embryonic death, foetal death, growth retardation, small head, low brain weight, micro-phthalmia and any other gross morphological abnormalities. The period of maximum sensitivity for each effect varied. The only demonstrable effect of irradiation during the pre-implantation period was an increase in prenatal mortality. Resorptions were maximal after exposure between days 2 and 4 p.c. The pre-implantation irradiated embryos which survived did not show any major foetal abnormalities. These results confirm earlier mouse studies using higher doses of X-rays. Small head, low brain weight and microphthalmia were prominent after exposure during the late organogenesis period, especially between days 9 and 13 p.c. But no other externally visible anomalies were detected. These findings demonstrate that mouse organogenesis is very sensitive to radiation-induced retardation of development, even at doses < 1 Gy. One exencephaly, one cleft palate and two cases of open eyelids were observed in the foetuses exposed on days 14.5 and 15.5 p.c.; the number of these cases was too small to indicate a causal relationship with exposure.

PURPOSE:To study the response of a mouse melanoma to radiation and hyperthermia under acute hypoxia and reperfusion.MATERIALS AND METHODS:B16F1 melanoma of 100+/-10 mm3, in C57BL mouse, were locally exposed to 10Gy gamma radiation (RT), 43 degrees C for 30 min (HT) in a water bath, or RT followed immediately by HT, under clamping (acute hypoxia) or 1 h after reperfusion. Tumour regression, volume doubling time (VDT), growth delay (GD), apoptosis and microvascular density (MVD) were studied.RESULTS:Under clamping, HT increased the VDT and GD to > 20 days above control and resulted in > 50% regression (PR) in all the tumours, whilst RT + HT synergistically enhanced VDT and GD. Under reperfusion, HT produced 25% PR against 16% by RT, with no increase in VDT and GD compared to RT. RT + HT significantly enhanced VDT and GD above that of RT or HT, but did not further increase PR of reperfused tumours. HT under clamping caused > 50% increase in apoptic cells over control and decreased MVD to 1/3rd of control. RT + HT further enhanced apoptotic cells to > 70% and reduced MVD to 1/6th of control.CONCLUSIONS:These results suggest that combination of radiotherapy with hyperthermia could benefit treatment of tumours with ischemia-induced acute hypoxia.

PURPOSE:To study long-term changes in the adult locomotor activity of mice after exposure to gamma radiation at the early fetal stage of development.MATERIALS AND METHODS:Pregnant Swiss albino mice were exposed locally on the abdominal area to a single dose of 0.25-1.5 Gy of 60Co gamma radiation at the dose rate of 1 Gy/min. When the F1 offspring were 6 months old, their locomotor and exploratory behaviour was assessed by the open-field and dark/bright arena tests. Animals were again subjected to the dark/bright arena test at 12 and 18 months of age in order to study the persistence of the effects.RESULTS:Irradiation produced a noticeable disturbance in the normal behaviour pattern of the mice. There was a significant dose-dependent decrease in the open-field activity of 6-month-old mice. In the dark/bright arena test, the time spent and lines crossed in the dark area showed a significant decrease, while their activities in the brightly lit area increased significantly, indicating a reduced aversion to bright light. These effects were evident even at a dose of 0.3 Gy and increased linearly with dose. The significant behavioural changes persisted at 12 months, but at 18 months the difference in the time spent and lines crossed in the dark and bright areas were not significantly different from sham-irradiated control values below 0.5 Gy.CONCLUSIONS:These results show that day 14 of gestation in Swiss albino mice is a time of high risk for inducing long-term changes in the adult locomotor function by gamma-radiation doses below 1 Gy. Using a range of radiation doses and different observation times we have demonstrated that the effect increases linearly with dose, but there appears to be a threshold of 0.3-0.5 Gy for producing significant persistent changes in the adult ambulatory activity.

In previous studies, flavonoids, orientin and vicenin, that were isolated from the leaf extract of Ocimum sanctum, were found to protect mice against radiation injury. Several flavonoids are known to be good antioxidants. Therefore, the effect of orientin and vicenin on radiation-induced lipid peroxidation in vivo and their antioxidant activity in vitro were studied. Adult mice were injected intraperitoneally with 50 microgram/kg of orientin or vicenin and exposed whole-body to 3 Gy of gamma radiation. Lipid peroxidation was measured in the liver 15 min to 8 h postirradiation. The antioxidant activity of orientin/vicenin (10-500 microM) was studied by measuring inhibition of hydroxyl radicals generated by the Fenton reaction (Fe(3+)-EDTA-ascorbic acid-H(2)O(2)) in vitro. The compounds were also tested for possible pro-oxidant and iron chelation activities at the above concentrations in the in vitro system. Orientin and vicenin provided almost equal protection against radiation-induced lipid peroxidation in mouse liver. Both compounds showed a significantly greater free radical-inhibiting activity in vitro than DMSO. Neither orientin nor vicenin showed any pro-oxidant activity at the concentrations tested. Both compounds inhibited free radical formation in the absence of EDTA. Free radical scavenging appears to be a likely mechanism of radiation protection by these flavonoids.

PURPOSE:To study the tumourigenic effect of prenatal low-dose gamma-irradiation in the mouse.METHODS AND MATERIALS:Pregnant Swiss albino mice were exposed to 0.1-1.5 Gy gamma-radiation on days 14 or 17 of gestation. The F1 offspring were observed up to 18 months of age. All the mice were killed at 18 months and the incidence of tumours in different organs was recorded.RESULTS:Exposure to doses from 0.1 to 1.5 Gy on days 14 or 17 of gestation produced a linear-quadratic dose-dependent increase in tumour incidence in adult F1 mice. The main organs affected were the ovary, uterus, liver and spleen. The highest incidence was observed in the ovaries, which was significantly higher than spontaneous incidence, even at 0.25 Gy. In other organs the tumour incidence was not significant compared with controls at doses < 0.5-1.0 Gy. Tumours in the ovary and uterus developed at an earlier age than in the liver and spleen.CONCLUSIONS:Exposure to gamma-radiation < 1.0 Gy at the foetal period (days 14 or 17 of gestation) can cause induction of tumours in the Swiss albino mouse. The carcinogenic effect, particularly on the ovary among the female mouse, is detectable after low-dose foetal irradiation.

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.

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.

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.

Micronuclei (MN) induction in the mouse bone marrow polychromatic erythrocytes (PCE) and normochromatic erythrocytes (NCE) was studied 24 h after whole body exposure to 4 Gy of gamma radiation with or without a single intraperitoneal injection of 25 mg/kg to 150 mg/kg of 50% methanolic extract of Phyllanthus niruri (whole aerial part). The extract significantly reduced the radiation-induced MN induction in both PCE and NCE and increased the ratio of PCE to NCE. The effect increased linearly with extract dose from 25 to 125 mg/kg.

UMA DEVI, P., M. HOSSAIN AND K. S. BISHT. Effect of late fetal irradiation on adult behavior of mouse: Dose–response relationship. NEUROTOXICOL TERATOL 21(2) 193–198, 1999.—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.

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.

Two flavonoids, orientin and vicenin, isolated from the leaves of the Indian plant Ocimum sanctum were tested for their radioprotective effect in mice. Both compounds provided protection against death from gastrointestinal syndrome as well as bone marrow syndrome when injected intraperitoneally (i.p.) before whole-body exposure to 11 Gy gamma radiation. The optimum drug dose for protection was 50 microg/kg body weight: An increase in the drug dose did not increase protection. No acute toxicity was observed at doses as high as 100 mg/kg body weight of either compound. Maximum protection was obtained when either compound was injected i.p. 30 min before irradiation. Changing the route of administration or the interval between drug injection (i.p.) and irradiation reduced protection. Drug treatment after irradiation was not very effective. Vicenin was slightly better than orientin in increasing survival at 30 days; protection by vicenin also lasted longer. Dose modification factors (DMFs) for the LD50 were 1.37 for vicenin and 1.30 for orientin. Radical scavenging activity has been demonstrated for both orientin and vicenin, and this appears to be one of the mechanisms of protection by these flavonoids.

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.

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.