Endothelin receptor antagonist improves donor lung function in an ex vivo perfusion system.

BACKGROUND: A lung transplant is the last resort treatment for many patients with advanced lung disease. The majority of donated lungs come from donors following brain death (BD). The endothelin axis is upregulated in the blood and lung of the donor after BD resulting in systemic inflammation, lung damage and poor lung graft outcomes in the recipient. Tezosentan (endothelin receptor blocker) improves the pulmonary haemodynamic profile; however, it induces adverse effects on other organs at high doses. Application of ex vivo lung perfusion (EVLP) allows the development of organ-specific hormone resuscitation, to maximise and optimise the donor pool. Therefore, we investigate whether the combination of EVLP and tezosentan administration could improve the quality of donor lungs in a clinically relevant 6-h ovine model of brain stem death (BSD). METHODS: After 6 h of BSD, lungs obtained from 12 sheep were divided into two groups, control and tezosentan-treated group, and cannulated for EVLP. The lungs were monitored for 6 h and lung perfusate and tissue samples were processed and analysed. Blood gas variables were measured in perfusate samples as well as total proteins and pro-inflammatory biomarkers, IL-6 and IL-8. Lung tissues were collected at the end of EVLP experiments for histology analysis and wet-dry weight ratio (a measure of oedema). RESULTS: Our results showed a significant improvement in gas exchange [elevated partial pressure of oxygen (P = 0.02) and reduced partial pressure of carbon dioxide (P = 0.03)] in tezosentan-treated lungs compared to controls. However, the lungs hematoxylin-eosin staining histology results showed minimum lung injuries and there was no difference between both control and tezosentan-treated lungs. Similarly, IL-6 and IL-8 levels in lung perfusate showed no difference between control and tezosentan-treated lungs throughout the EVLP. Histological and tissue analysis showed a non-significant reduction in wet/dry weight ratio in tezosentan-treated lung tissues (P = 0.09) when compared to control. CONCLUSIONS: These data indicate that administration of tezosentan could improve pulmonary gas exchange during EVLP.

In vitro inhibition activity of essential oils from some Lamiaceae species against phytopathogenic fungi.

Natural products have been in focus as alternative, effective and safe materials against the phytopathogens. Investigations show Nepeta oils as effective in controlling the food crops decay. The inhibitory effects of essential oils derived from Nepeta leucophylla, Nepeta ciliaris, Nepeta clarkei and Calamintha umbrosa against five phytopthogenic fungi have been determined. In vitro antifungal activity varied with their constituents and target species. More active being the oils containing oxygenated terpenoids. Helminthosporium maydis was sensitive to the all oils, IC50 values have 43.6-109.3 µg mL(-1). The N. leucophylla oil possessing oxygenated iridoids was more effective against H. maydis (IC50 value of 43.6 µg mL(-1)) while N. ciliaris was more active against Fusarium oxysporum (IC50 value of 219.2 µg mL(-1)). The oils were effective against the spore germination of all the tested plant pathogens.

Brain stem death induces pro-inflammatory cytokine production and cardiac dysfunction in sheep model.

INTRODUCTION: Organs procured following brain stem death (BSD) are the main source of organ grafts for transplantation. However, BSD is associated with inflammatory responses that may damage the organ and affect both the quantity and quality of organs available for transplant. Therefore, we aimed to investigate plasma and bronchoalveolar lavage (BAL) pro-inflammatory cytokine profiles and cardiovascular physiology in a clinically relevant 6-h ovine model of BSD. METHODS: Twelve healthy female sheep (37-42 Kg) were anaesthetized and mechanically ventilated prior to undergoing BSD induction and then monitored for 6 h. Plasma and BAL endothelin-1 and cytokines (IL-1ß, 6, 8 and tumour necrosis factor alpha (TNF-α)) were assessed by ELISA. Differential white blood cell counts were performed. Cardiac function during BSD was also examined using echocardiography, and cardiac biomarkers (A-type natriuretic peptide and troponin I were measured in plasma. RESULTS: Plasma concentrations big ET-1, IL-6, IL-8, TNF-α and BAL IL-8 were significantly (p < 0.01) increased over baseline at 6 h post-BSD. Increased numbers of neutrophils were observed in the whole blood (3.1 × 109 cells/L [95% confidence interval (CI) 2.06-4.14] vs. 6 × 109 cells/L [95%CI 3.92-7.97]; p < 0.01) and BAL (4.5 × 109 cells/L [95%CI 0.41-9.41] vs. 26 [95%CI 12.29-39.80]; p = 0.03) after 6 h of BSD induction vs baseline. A significant increase in ANP production (20.28 pM [95%CI 16.18-24.37] vs. 78.68 pM [95%CI 53.16-104.21]; p < 0.0001) and cTnI release (0.039 ng/mL vs. 4.26 [95%CI 2.69-5.83] ng/mL; p < 0.0001), associated with a significant reduction in heart contractile function, were observed between baseline and 6 h. CONCLUSIONS: BSD induced systemic pro-inflammatory responses, characterized by increased neutrophil infiltration and cytokine production in the circulation and BAL fluid, and associated with reduced heart contractile function in ovine model of BSD.

Essential omega-3 fatty acids tune microglial phagocytosis of synaptic elements in the mouse developing brain.

Omega-3 fatty acids (n-3 PUFAs) are essential for the functional maturation of the brain. Westernization of dietary habits in both developed and developing countries is accompanied by a progressive reduction in dietary intake of n-3 PUFAs. Low maternal intake of n-3 PUFAs has been linked to neurodevelopmental diseases in Humans. However, the n-3 PUFAs deficiency-mediated mechanisms affecting the development of the central nervous system are poorly understood. Active microglial engulfment of synapses regulates brain development. Impaired synaptic pruning is associated with several neurodevelopmental disorders. Here, we identify a molecular mechanism for detrimental effects of low maternal n-3 PUFA intake on hippocampal development in mice. Our results show that maternal dietary n-3 PUFA deficiency increases microglia-mediated phagocytosis of synaptic elements in the rodent developing hippocampus, partly through the activation of 12/15-lipoxygenase (LOX)/12-HETE signaling, altering neuronal morphology and affecting cognitive performance of the offspring. These findings provide a mechanistic insight into neurodevelopmental defects caused by maternal n-3 PUFAs dietary deficiency.

Pot1 OB-fold mutations unleash telomere instability to initiate tumorigenesis.

Chromosomal aberrations are a hallmark of human cancers, with complex cytogenetic rearrangements leading to genetic changes permissive for cancer initiation and progression. Protection of Telomere 1 (POT1) is an essential component of the shelterin complex and functions to maintain chromosome stability by repressing the activation of aberrant DNA damage and repair responses at telomeres. Sporadic and familial mutations in the oligosaccharide-oligonucleotide (OB) folds of POT1 have been identified in many human cancers, but the mechanism underlying how hPOT1 mutations initiate tumorigenesis has remained unclear. Here we show that the human POT1's OB-folds are essential for the protection of newly replicated telomeres. Oncogenic mutations in hPOT1 OB-fold fail to bind to single-stranded telomeric DNA, eliciting a DNA damage response at telomeres that promote inappropriate chromosome fusions via the mutagenic alternative non-homologous end joining (A-NHEJ) pathway. hPOT1 mutations also result in telomere elongation and the formation of transplantable hematopoietic malignancies. Strikingly, conditional deletion of both mPot1a and p53 in mouse mammary epithelium resulted in development of highly invasive breast carcinomas and the formation of whole chromosomes containing massive arrays of telomeric fusions indicative of multiple breakage-fusion-bridge cycles. Our results reveal that hPOT1 OB-folds are required to protect and prevent newly replicated telomeres from engaging in A-NHEJ mediated fusions that would otherwise promote genome instability to fuel tumorigenesis.

Chromosome damage and micronucleus formation in human blood lymphocytes exposed in vitro to radiofrequency radiation at a cellular telephone frequency (847.74 MHz, CDMA).

Peripheral blood samples collected from four healthy nonsmoking human volunteers were diluted with tissue culture medium and exposed in vitro for 24 h to 847.74 MHz radiofrequency (RF) radiation (continuous wave), a frequency employed for cellular telephone communications. A code division multiple access (CDMA) technology was used with a nominal net forward power of 75 W and a nominal power density of 950 W/m(2) (95 mW/cm(2)). The mean specific absorption rate (SAR) was 4.9 or 5.5 W/kg. Blood aliquots that were sham-exposed or exposed in vitro to an acute dose of 1.5 Gy of gamma radiation were included in the study as controls. The temperatures of the medium during RF-radiation and sham exposures in the Radial Transmission Line facility were controlled at 37 +/- 0.3 degrees C. Immediately after the exposures, lymphocytes were cultured at 37 +/- 1 degrees C for 48 or 72 h. The extent of genetic damage was assessed from the incidence of chromosome aberrations and micronuclei. The kinetics of cell proliferation was determined from the mitotic indices in 48-h cultures and from the incidence of binucleate cells in 72-h cultures. The data indicated no significant differences between RF-radiation-exposed and sham-exposed lymphocytes with respect to mitotic indices, frequencies of exchange aberrations, excess fragments, binucleate cells, and micronuclei. The response of gamma-irradiated lymphocytes was significantly different from that of both RF-radiation-exposed and sham-exposed cells for all of these indices. Thus there was no evidence for induction of chromosome aberrations and micronuclei in human blood lymphocytes exposed in vitro for 24 h to 847.74 MHz RF radiation (CDMA) at SARs of 4.9 or 5.5 W/kg.

Cytogenetic studies in human blood lymphocytes exposed in vitro to radiofrequency radiation at a cellular telephone frequency (835.62 MHz, FDMA).

Freshly collected peripheral blood samples from four healthy human volunteers were diluted with RPMI 1640 tissue culture medium and exposed in sterile T-75 tissue culture flasks in vitro for 24 h to 835.62 MHz radiofrequency (RF) radiation, a frequency employed for customer-to-base station transmission of cellular telephone communications. An analog signal was used, and the access technology was frequency division multiple access (FDMA, continuous wave). A nominal net forward power of 68 W was used, and the nominal power density at the center of the exposure flask was 860 W/m(2). The mean specific absorption rate in the exposure flask was 4.4 or 5.0 W/kg. Aliquots of diluted blood that were sham-exposed or exposed in vitro to an acute dose of 1.50 Gy of gamma radiation were used as negative or positive controls. Immediately after the exposures, the lymphocytes were stimulated with a mitogen, phytohemagglutinin, and cultured for 48 or 72 h to determine the extent of genetic damage, as assessed from the frequencies of chromosomal aberrations and micronuclei. The extent of alteration in the kinetics of cell proliferation was determined from the mitotic indices in 48-h cultures and from the incidence of binucleate cells in 72-h cultures. The data indicated no significant differences between RF-radiation- and sham-exposed lymphocytes with respect to mitotic indices, incidence of exchange aberrations, excess fragments, binucleate cells, and micronuclei. In contrast, the response of the lymphocytes exposed to gamma radiation was significantly different from both RF-radiation- and sham-exposed cells for all of these indices. Thus, under the experimental conditions tested, there is no evidence for the induction of chromosomal aberrations and micronuclei in human blood lymphocytes exposed in vitro for 24 h to 835.62 MHz RF radiation at SARs of 4.4 or 5.0 W/kg.

Neoplastic transformation in C3H 10T(1/2) cells after exposure to 835.62 MHz FDMA and 847.74 MHz CDMA radiations.

The effect of radiofrequency (RF) radiation in the cellular phone communication range (835.62 MHz frequency division multiple access, FDMA; 847.74 MHz code division multiple access, CDMA) on neoplastic transformation frequency was measured using the in vitro C3H 10T(1/2) cell transformation assay system. To determine if 835.62 MHz FDMA or 847.74 MHz CDMA radiations have any genotoxic effects that induce neoplastic transformation, C3H 10T(1/2) cells were exposed at 37 degrees C to either of the above radiations [each at a specific absorption rate (SAR) of 0.6 W/kg] or sham-exposed at the same time for 7 days. After the culture medium was changed, the cultures were transferred to incubators and refed with fresh growth medium every 7 days. After 42 days, the cells were fixed and stained with Giemsa, and transformed foci were scored. To determine if exposure to 835.62 MHz FDMA or 847.74 MHz CDMA radiation has any epigenetic effects that can promote neoplastic transformation, cells were first exposed to 4.5 Gy of X rays to induce the transformation process and then exposed to the above radiations (SAR = 0.6 W/kg) in temperature-controlled irradiators with weekly refeeding for 42 days. After both the 7-day RF exposure and the 42-day RF exposure after X irradiation, no statistically significant differences in the transformation frequencies were observed between incubator controls, the sham-exposed (maintained in irradiators without power to the antenna), and the 835.62 MHz FDMA or 847.74 MHz CDMA-exposed groups.

Measurement of DNA damage in mammalian cells exposed in vitro to radiofrequency fields at SARs of 3-5 W/kg.

In the present study, we determined whether exposure of mammalian cells to 3.2-5.1 W/kg specific absorption rate (SAR) radiofrequency fields could induce DNA damage in murine C3H 10T(1/2) fibroblasts. Cell cultures were exposed to 847.74 MHz code-division multiple access (CDMA) and 835.62 frequency-division multiple access (FDMA) modulated radiations in radial transmission line (RTL) irradiators in which the temperature was regulated to 37.0 +/- 0.3 degrees C. Using the alkaline comet assay to measure DNA damage, we found no statistically significant differences in either comet moment or comet length between sham-exposed cells and those exposed for 2, 4 or 24 h to CDMA or FDMA radiations in either exponentially growing or plateau-phase cells. Further, a 4-h incubation after the 2-h exposure resulted in no significant changes in comet moment or comet length. Our results show that exposure of cultured C3H 10T(1/2) cells at 37 degrees C CDMA or FDMA at SAR values of up to 5.1 W/kg did not induce measurable DNA damage.

Constituents of the yew trees.

Yew trees, taxonomically classified under the genus Taxus, are sources of a number of physiologically active compounds of different classes. Taxane derivatives with various carbon skeletons, lignans, flavonoids, steroids and sugar derivatives have been isolated from different Taxus species. Compounds isolated from the genus Taxus between 1908 and December 1997 have been comprehensively reviewed.

Effect of gamma radiation on the foetal haemopoietic system in the mouse.

PURPOSE: To study the effect of gamma irradiation, in the late foetal period, on the haemopoietic progenitor cells of the mouse foetus. MATERIALS AND METHODS: Pregnant Swiss mice were exposed to 0.1 to 1.5 Gy of Cobalt-60 gamma radiation on the 17th day of gestation. At 24h (18 day p.c.) and 72 h (20 day p.c.) after exposure the weights of foetal liver and spleen and their mean cellularity were determined. Chromosomal aberrations and micronuclei (MN) were studied at 24 h. Haemopoietic progenitor cell survival at 24 and 72 h post-irradiation was studied by exogenous spleen colony assay. RESULTS: Irradiation caused significant reduction in the weights of foetal liver and spleen, which was more pronounced at 72 h post-irradiation. The mean cellularity as well as the CFU-S8 and CFU-S12 of liver and spleen showed a radiation dose-dependent decrease, but the spleen showed a higher sensitivity than liver to doses below 1.0 Gy. Chromosomal damage in liver increased linearly with dose, while in spleen the increase was linear-quadratic. CONCLUSIONS: The results demonstrate the damaging effect of low doses of gamma radiation on the late foetal haemopoietic progenitor cells of the mouse. Induction of cytogenetic damage appears to be a no-threshold effect in the dose range used.

Inhibition of cyclooxygenase-2 with NS-398 and the prevention of radiation-induced transformation, micronuclei formation and clonogenic cell death in C3H 10T1/2 cells.

PURPOSE: Abnormally high levels of the cyclooxygenase (COX)-2 isozyme as well as the prostaglandin metabolites produced by the COX pathway have been observed in a variety of malignancies, including cancers of the skin, pancreas, colon, breast, cervix, prostate, and head and neck. Furthermore, exogenous genotoxic agents, including ionizing radiation (IR), have been shown to induce cellular transformation and to elevate COX-2 activity, whereas exposure to agents that specifically inhibit COX-2 activity have been shown to inhibit transformation. These data suggest a possible role of COX-2 both in IR-mediated cellular transformation processes and cell death. MATERIALS AND METHODS: C3H 10T1/2 and/or HeLa cells were treated with N-[2-(cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide (NS-398) and/or exposed to IR. Following treatment, cells were assayed for neoplastic transformation, clonogenicity, growth rates, cell cycle distribution, micronuclei formation and DNA damage by established methodologies. Statistical tests were performed on data as described. RESULTS: In the present study, experiments in normal murine fibroblast C3H 10T1/2 cells demonstrated that the chemical inhibition of COX-2 activity with moderate doses of NS-398 abrogated IR-induced transformation events by fourfold and protected irradiated C3H 10T1/2 cells from clonogenic cell death. Considering that these doses of NS-398 had no significant effect on cellular proliferation or cell cycle distribution in C3H 10T1/2 cells, the results suggest that inhibition of COX-2 either increases DNA repair or prevents the accumulation of DNA damage. In supplemental experiments, treatment with NS-398 caused a 1.5-fold reduction in IR-induced micronuclei formation and a significant decrease in DNA damage. CONCLUSIONS: These results suggest a role for COX-2 inhibitors in the normal tissue response to IR when administered at therapeutically achievable doses and therefore may have clinical implications for radiation oncology patients in the prevention of IR-induced malignancy.

Micronuclei in the peripheral blood and bone marrow cells of rats exposed to 2450 MHz radiofrequency radiation.

PURPOSE: To determine the incidence of micronuclei in peripheral blood and bone marrow cells of rats exposed continuously for 24h to 2450 MHz continuous wave radiofrequency radiation (RFR) at an average whole-body specific absorption rate (SAR) of 12W/kg. MATERIALS AND METHODS: Eight adult male Sprague-Dawley rats were exposed to 2450 MHz RFR in circularly polarized waveguides. Eight sham-exposed rats were kept in similar waveguides without the transmission of RFR. Four rats were treated with mitomycin-C (MMC) and used as positive controls. All rats were necropsied 24h after the end of RFR and sham exposures, and after the 24h treatment with MMC. Peripheral blood and bone marrow smears were examined to determine the frequency of micronuclei (MN) in polychromatic erythrocytes (PCE). RESULTS: The results indicated that the incidence of MN/2000 PCE were not significantly different between RFR- and sham-exposed rats. The group mean frequencies of MN in the peripheral blood were 2.3+/-0.7 in RFR-exposed rats and 2.1+/-0.6 in sham-exposed rats. In bone marrow cells, the average MN incidence was 3.8+/-1.0 in RFR-exposed rats and 3.4+/-0.7 in sham-exposed rats. The corresponding values in positive control rats treated with MMC were 23.5+/-4.7 in the peripheral blood and 33.8+/-7.4 in bone marrow cells. CONCLUSION: There was no evidence for the induction of MN in peripheral blood and bone marrow cells of rats exposed for 24h to 2450 MHz continuous wave RFR at a whole body average SAR of 12 W/kg.

Dose-dependent increase in the frequency of micronuclei and chromosomal aberrations by misonidazole in mouse bone marrow.

Cytogenetic effects produced in bone marrow of mice by various doses of misonidazole (MISO, 100 to 1000 mg/kg bodyweight) were studied by assaying the induction of micronuclei (MN) and chromosomal aberrations. Misonidazole increased the frequency of both micronuclei and chromosomal aberrations over normal at 24 h after treatment, however, the values were statistically significant from normal only at drug doses above 250 mg/kg. The increase was proportional to the drug dose with a best fit to linear quadratic model for MN induction. For chromosomal aberrations the data fitted equally well to linear as well as linear quadratic models.

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.

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.

Hyperthermia in cancer research: current status.

There are critical maximum temperatures above which irreversible damage occurs in cells and tissues. Exposure to high temperature, referred to as hyperthermia (HT), can result in cell death, tissue damage or even death of the organism. Clinical application of HT as a primary treatment or as an adjuvant to radio-/chemo- therapy of cancer is based on its ability to cause localized tumor tissue damage. Experimental data provide HT with a strong biological rationale. Early clinical experience suggested that HT will become an important modality as an adjuvant to radiotherapy in the treatment of human malignancies, but its application is currently limited to mainly superficial tumors. Its full realization as a treatment modality for cancer therapy awaits further laboratory investigations as well as controlled clinical trials. A better understanding of the biological mechanisms of its action, interaction with chemotherapeutic drugs and radiation damage, role of tumor microenvironment such as oxygen status and pH of tumors, and kinetics of thermotolerance can lead to refinement in its clinical implementation. The present review attempts to analyse the published literature during the last one and half decades.

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.

Effect of gamma radiation on fetal haemopoiesis of mouse.

Effect of a single acute exposure to gamma-rays during the late fetal period on the fetal haemopoietic tissue of mouse was studied. Pregnant Swiss albino mice were exposed to 1 Gy of gamma-rays on day 17 post coitus (late fetal period) and 24 hr after exposure the fetuses were observed for changes in the liver weight, cytogenetic damage in liver cells by micronucleus (MN) induction and stem cell survival by spleen colony (CFU-S) assay. Irradiation resulted in a significant (P < 0.01) decrease in fetal liver weight. A significant (P < 0.001) increase in MN count was observed after exposure, while CFU-S displayed a significant (P < 0.001) reduction in the cell survival as compared to the sham-treated control. These results demonstrate the high susceptibility of mouse fetal haemopoietic system to radiation.

Differentiation of mouse embryonal carcinoma cells PCC4 by heat shock and the kinetics of induction of heat shock proteins.

Heat shock to embryonal carcinoma cells PCC4 at 45 degrees C for 30 min resulted in the differentiation of cells although heat shock response was induced on exposure to 42 degrees C for 60 min. Differentiated cells were large and well spread with reduced nuclear/cytoplasmic ratios as compared to undifferentiated cells. Change in cell morphology was associated with the disappearance and appearance of stage specific embryonic antigens 1 and 3 respectively. We also found a change in intracellular pH in PCC4 cells within 30 min of heat shock as measured by the change in fluorescence intensity of a probe incorporated into cells during heat shock.