Proliferation and differentiation of neural stem cells irradiated with X-rays in logarithmic growth phase

Exposure of the fetal brain to ionizing radiation causes congenital brain abnormalities. Normal brain formation requires regionally and temporally appropriate proliferation and differentiation of neural stem cells (NSCs) into neurons and glia. Here, we investigated the effects of X-irradiation on proliferating homogenous NSCs prepared from mouse ES cells. Cells irradiated with X-rays at a dose of 1Gy maintained the capabilities for proliferation and differentiation but stopped proliferation temporarily. In contrast, the cells ceased proliferation following irradiation at a dose of >5Gy. These results suggest that irradiation of the fetal brain at relatively low doses may cause congenital brain abnormalities as with relatively high doses.     

UVA1 irradiation inhibits fibroblast proliferation and alleviates pathological changes of scleroderma in a mouse model

The purpose of the present study was to compare the effects of different doses of ultraviolet radiation A1 (UVA1) on human fibroblast proliferation and collagen level in a mouse model of scleroderma,so as to identify appropriate irradiation doses for clinical treatment of scleroderma.Monolayer from human fibroblasts was cultured in vitro,and a mouse model of scleroderma was established by subcutaneous injection of 100 μL of 400 μg/mL bleomycin into the back of BALB/c mice for 4 weeks.The mouse models and human fibroblasts were divided into UVA1exposed (100,60 and 20 J/cm 2) and UVA-unexposed groups.At 0,24 and 48 h after exposure,cell proliferation and levels of hydroxyproline and collagen were detected.UVA1 irradiation was performed 3 times weekly for 10 weeks,and the pathological changes of skin tissues,skin thickness and collagen level were observed after phototherapy.Cell proliferation and the levels of hydroxyproline and collagen were inhibited after phototherapy,and there was a significant difference between the UVA1-exposed cells and UVA1-unexposed cells (P < 0.001).In addition,UVA1 phototherapy improved dermal sclerosis and softened the skin,and there were significant differences between the high-dose UVA1 group and the model group,and the negative group (P < 0.05).It is concluded that UVA1 radiation can reduce cell proliferation,and decrease hydroxyproline and collagen levels in a dose-dependent manner in vitro.High-dose UVA1 phototherapy has marked therapeutic effect on scleroderma in the mouse model.Decreased collagen level may be related to the reduced number and activity of cells,as well as inhibition of collagen synthesis.     

二甲苯对妊娠小鼠及胚胎发育的毒性作用

目的探讨二甲苯对妊娠小鼠及胚胎发育的影响.方法对妊娠初期的雌性小鼠采用静式吸入染毒法进行二甲苯染毒,观察胎鼠体重变化;原子吸收分光光度测定法（atomic absorption spectroscopy,AAS）检测妊娠母鼠血清Fe^2＋、Mg^2＋含量;单细胞凝胶电泳法（single cell gel electrophoresis,SCGE）检测妊娠母鼠外周血有核细胞DNA损伤程度.结果染毒各组妊娠母鼠血清Fe^2＋、Mg^2＋含量下降（P＜0.01,P＜0.05）,并具有统计学意义;染毒组妊娠母鼠外周血有核细胞发生DNA断裂并形成彗尾图像;染毒各组胎鼠体重较正常组相比均显著下降,且存在显著性差异（P＜0.05）;部分胎鼠表现为胚胎被吸收、脑膜脑膨出、椎骨发育不全等.结论二甲苯可造成妊娠母鼠血清Fe^2＋、Mg^2＋含量下降、外周血有核细胞DNA损伤并致使流产发生;二甲苯引起胎鼠体重下降,并影响生长发育.     

Aggregation patterns of fetal rat brain cells following exposure to X-irradiation

In our search for a simplified in vitro test system to assess the teratogenic effects of physical factors, we studied the effects of total maternal body X-irradiation on aggregation patterns of enzymatically isolated fetal rat brain cells and on ultrastructural aggregate changes. The fetal brain cells were derived from day 14 gestation fetuses of pregnant Sprague-Dawley (CD strain) rats exposed to X-irradiation (25 - 200 R) one hour prior to sacrifice. Notable changes in the cell aggregates following X-irradiation included a reduction in cell aggregate size and an increase in number. The frequency of cell aggregates was higher in the treated than in the control group, and the mean diameter of cell aggregates was inversely related to increasing X-irradiation doses. Transmission electron microscopy revealed in isolated cells features of degenerative process which were similar to those found in intact fetal brain lesions caused by maternal X-irradiation. Furthermore, scanning electron microscopy revealed that inhibition of cell aggregation following X-irradiation could probably be attributed to inhibition of membrane filopodia development and a consequent failure fo cell aggregates to fuse into a greater cell aggregate mass. These results suggest that the membrane factors which influence cell aggregation may be a useful parameter to assess early effects of X-irradiation-induced brain deformity. Presently, the cell aggregation culture system is being further evaluated as a short term test system for environmental teratogens.     

A feasibility study to calculate unshielded fetal doses to pregnant patients in 6-MV photon treatments using Monte Carlo methods and anatomically realistic phantoms

A Monte Carlo-based procedure to assess fetal doses from 6-MV external photon beam radiation treatments has been developed to improve upon existing techniques that are based on AAPM Task Group Report 36 published in 1995 [M. Stovall et al., Med. Phys. 22, 63-82 (1995)]. Anatomically realistic models of the pregnant patient representing 3-, 6-, and 9-month gestational stages were implemented into the MCNPX code together with a detailed accelerator model that is capable of simulating scattered and leakage radiation from the accelerator head. Absorbed doses to the fetus were calculated for six different treatment plans for sites above the fetus and one treatment plan for fibrosarcoma in the knee. For treatment plans above the fetus, the fetal doses tended to increase with increasing stage of gestation. This was due to the decrease in distance between the fetal body and field edge with increasing stage of gestation. For the treatment field below the fetus, the absorbed doses tended to decrease with increasing gestational stage of the pregnant patient, due to the increasing size of the fetus and relative constant distance between the field edge and fetal body for each stage. The absorbed doses to the fetus for all treatment plans ranged from a maximum of 30.9 cGy to the 9-month fetus to 1.53 cGy to the 3-month fetus. The study demonstrates the feasibility to accurately determine the absorbed organ doses in the mother and fetus as part of the treatment planning and eventually in risk management.     

The effects of prenatal X-irradiation on hypoglossal nucleus: a GFAP immunohistochemical study

The effects of prenatal X-irradiation on hypoglossal (XII) nucleus were investigated in the rat. Pregnant animals were exposed to a single whole body X-irradiation on day 11 and 16 of gestation at a does of 1.3 Gy. The offspring were killed at 7-14 days of age for the histological and immunohistochemical observations. Nissl staining revealed no significant changes of XII motoneurons in these experimental animals. In the control case it was of interest that expression of glial fibrillary acidic protein-immunoreactivity (GFAP-IR) is largely confined to the dorsomedial region including the XII nucleus at the level caudal to the obex. Exposure of X-irradiation on day 16 of gestation led to similar expression of GFAP-IR in the nucleus at the same level. However, exposure on day 11 of gestation apparently led to strong expression of GFAP-IR in the XII nucleus at the level caudal to the obex and the expression was observed to extend rostrally. The GFAP-IR cells showed hypertrophy of cell bodies and longer cytoplasmic processes. Horse-radish peroxidase (HRP) injection into the tip of the tongue including the intrinsic muscles resulted in retrograde labeling in the ventromedial portion of the XII nucleus bilaterally from +0.30 to -1.25 mm. The present study would indicate that motoneurons of the XII nucleus supplying mainly the intrinsic and partly the extrinsic tongue muscles are more sensitive to X-ray exposure before the formation of the XII nucleus.     

Multiple abnormalities in the ultraviolet light response of cultured fibroblasts derived from patients with the basal cell nevus syndrome

Basal Cell Nevus Syndrome (BCNS) is a rare autosomal-dominant inherited disorder associated with a marked hypersusceptibility to spontaneous and radiation-induced skin cancer. We examined the changes in cell survival, unscheduled DNA synthesis (UDS) and the frequency of sister chromatid exchanges (SCE) induced by ultraviolet light (UVL) in confluent normal and BCNS fibroblasts. BCNS cells appeared slightly hypersensitive to the cytotoxic effects of UVL. The rate of UDS induced by UVL exposure in normal cell strains increased linearly following doses up to 30 J/m2, whereas in BCNS cells UDS became saturated at doses of 10 J/m2 showing no further increase with doses up to 30 J/m2. UDS activity persisted for longer periods after UVL exposure in BCNS as compared with normal cells. The dose-response relationship for UVL-induced SCE was similar in normal and BCNS fibroblasts. However, the frequencies of UVL-induced SCE declined to near background levels in normal cells following 12-24 hr of confluent holding prior to subculture whereas they remained elevated in BCNS cells with holding times up to 24 hr after UVL exposure. Overall, these results suggest that BCNS fibroblasts may have a diminished capacity for the repair of some type of DNA damage as compared with normal fibroblasts.     

Correlations between the degree and type of forebrain malformations and the simultaneous neuro-oncogenic properties of ethylnitrosourea after diaplacental exposure in rats, alone and in combination with X-irradiation

Single and combined treatments were performed in rats on day 13 of gestation with either ENU or ENU subsequent to various X-irradiation doses between 0.5 and 1.5 Gy. At this time of gestation, developmental anomalies of the brain are still inducible by any of these treatments, in addition to neurocarcinogenic effects after ENU alone or in combination with X-irradiation. We looked for correlations between the degree of brain malformations still detectable in the adult animals and the simultaneous occurrence of brain tumors. These evaluations were based on a histopathological analysis regarding the type and degree of malformation residues, as well as the type and distribution pattern of the tumors (especially regarding gliomas) within the forebrain. Both after ENU and X-irradiation plus ENU-treatment, the occurrence of glioma in the offspring was positively correlated with the degree of brain dysplasia. This effect was not only restricted to the total glioma incidence but also confirmed by the higher glioma multiplicity in major dysplastic brains. Additionally, gliomas were preferentially located within the subependymal layer, which simultaneously was most severely affected by the teratogenic effects after prenatal treatment. Although forebrain dysplasia generally presents a significant predisposition for glioma inducibility, this oncogenic event is apparently strictly inversely related to a certain type of forebrain malformation, namely the occurrence of heterotopic neuronal nodules within the telencephalic roof. They emerge from "rosettes," which are typical radiation lesions occurring only after doses above 1.0 Gy. In none of the forebrains which still revealed rosette-residues in later life could a simultaneous occurrence of gliomas be observed. This explains not only the substantial decrease of glioma incidence after combined treatment with 1.0 and 1.5 Gy X-irradiation doses, but also the consistent glioma multiplicity despite the decrease of frequency.     

Study of the effects of 0.15 terahertz radiation on genome integrity of adult fibroblasts

The applications of Terahertz (THz) technologies have significantly developed in recent years, and the complete understanding of the biological effects of exposure to THz radiation is becoming increasingly important. In a previous study, we found that THz radiation induced genomic damage in fetal fibroblasts. Although these cells demonstrated to be a useful model, exposure of human foetuses to THz radiation is highly improbable. Conversely, THz irradiation of adult dermal tissues is cause of possible concern for some professional and nonprofessional categories. Therefore, we extended our study to the investigation of the effects of THz radiation on adult fibroblasts (HDF). In this work, the effects of THz exposure on HDF cells genome integrity, cell cycle, cytological ultrastructure and proteins expression were assessed. Results of centromere‐negative micronuclei frequencies, phosphorylation of H2AX histone, and telomere length modulation indicated no induction of DNA damage. Concordantly, no changes in the expression of proteins associated with DNA damage sensing and repair were detected. Conversely, our results showed an increase of centromere‐positive micronuclei frequencies and chromosomal nondisjunction events, indicating induction of aneuploidy. Therefore, our results indicate that THz radiation exposure may affect genome integrity through aneugenic effects, and not by DNA breakage. Our findings are compared to published studies, and possible biophysical mechanisms are discussed. Environ. Mol. Mutagen. 59:476–487, 2018. © 2018 Wiley Periodicals, Inc.     

Induction of the putative protective protein ferritin by infrared radiation: implications in skin repair

The modification of ferritin in human skin cells in vitro and in vivo following infrared-A irradiation by immunohistochemical analysis and ELISA were evaluated. In addition, we observed that IR-A is not capable of inducing frank damage to DNA (pyrimidine dimers, p53), induction of oxidative stress proteins (heme oxygenase, nitric oxide, superoxide dismutase, heat shock proteins) or proteases (collagenase, stromelysin, gelatinase) involved in carcinogenesis and photoaging of the skin. in vivo, basal levels of ferritin were heterogeneous for all individuals tested but all showed ferritin to stain precisely in the basal layer of unirradiated epidermis. Following IR-A radiation, the ferritin increase was localized to epidermal tissue and showed an increase from 120 to 220%. Parallel to the in vivo analysis, dermal fibroblasts were cultured from six individuals. Quantitative analysis for ferritin in cultured fibroblasts was assessed by ELISA and increases were seen to be dose-dependent and up to 130% of basal levels of ferritin following infrared-A. Our findings indicate that the putative defense system of ferritin that exists in human skin in vivo can be induced by infrared-A radiation and that these wavelengths may prove to be beneficial for human skin. Importantly, following the same doses of IR-A that induced ferritin levels, there was no alteration seen for nuclear DNA type damage, oxidative stress proteins or proteases involved in the degradation of skin. The increased concentrations of this antioxidant in human skin following acute UV radiation could afford increased protection against subsequent oxidative stress.     

Recovery of glucocorticoid-related loss of synaptic density in the fetal sheep brain at 0.75 of gestation

Antenatal glucocorticoids routinely used to accelerate fetal lung maturation in human pregnancy at risk of preterm delivery decrease synaptic density and complex electrocortical activity in the fetal sheep brain at 0.87 gestation. We examined whether the effects of betamethasone on synaptic density depend on maturation of hypothalamo-pituitary-adrenal (HPA) axis and whether these effects are reversible. Betamethasone infusion to fetal sheep comparable to the dose used clinically (3.3 microg kg(-1) h(-1) over 48 h) at 0.75 gestation and, thus, before the prepartum increase of cortisol, reduced synaptophysin immunoreactivity (SY-IR) in the frontal neocortex, caudate putamen and hippocampus (P < 0.05). Loss of SY-IR exceeded that shown previously at 0.87 gestation (P < 0.05). It was not accompanied by neuronal damage and was reversible within 24h. In conclusion, fetal betamethasone exposure induces a gestational age-dependent decrease of synaptic density that is transient and more severe in younger fetuses.     

Genetic and teratogenic effects of cancer treatments on gametes and embryos

Male and female germ cells vary in their sensitivity to the mutagenic effects of chemotherapy and radiotherapy, depending on their stage of maturation and the agent used. Although sperm DNA damage exists following treatment, no increase in genetic defects or congenital malformations was detected among children conceived to parents who have previously undergone chemotherapy or radiotherapy. The use of assisted reproductive technologies and micromanipulation techniques might increase this risk; hence caution should be exercised. In female cancer patients, miscarriage and congenital malformations are not increased following chemotherapy. However, when IVF and embryo cryopreservation is practised between or shortly after treatment, possible genetic risks to the growing oocytes exist, and hence the babies should be screened. During pregnancy, the potential teratogenic effects of chemotherapy influence the choice and timing of therapy. Termination is usually recommended in the first trimester. Second- and third-trimester exposure does not usually increase the teratogenic risk and cognitive development, but it may increase the risk of poor obstetric outcome and fetal myelosuppression. During the first two weeks after fertilization of the embryo, radiation is lethal but not teratogenic. High doses of radiation during pregnancy induce anomalies, impaired growth and mental retardation, and there may be an increased risk of childhood leukaemia and other tumours in the offspring.     

Benzene-induced micronuclei formation in mouse fetal liver blood, peripheral blood, and maternal bone marrow cells

The transplacental cytogenetic effects of benzene were studied by using the micronucleus test of polychromatic erythrocytes (PCE) found in both fetal liver and fetal peripheral blood, and were compared with PCE from maternal bone marrow. Timed-pregnant mice received single intraperitoneal doses of benzene (0, 109, 219, 437, or 874 mg/kg bw) on the 14th day of gestation and were sacrificed 21 hr after injection. Benzene elicited a significant increase (P less than 0.01) in the frequency of micronucleated polychromatic erythrocytes (MNPCE) in fetal liver blood cells (0.55 to 1.36%, control 0.18%) at doses of 219 to 874 mg/kg, and in fetal peripheral blood cells (0.49 to 0.58%, control 0.25%) and maternal bone marrow cells (0.53 to 0.70%, control 0.10%) at doses of 437 and 874 mg/kg. The data demonstrate that benzene is a moderate transplacental clastogenic agent, and that the mouse transplacental micronucleus test using fetal liver blood cells is a potentially more sensitive indicator of the genotoxicity of benzene than either fetal peripheral blood or maternal bone marrow cells.     

Cutaneous injury following acute UV-B radiation in a mouse model: a pilot histological study

Abstract

Epidemiological and clinical studies have implicated solar ultraviolet (UV) radiation, especially the UV-B radiation (290–320 nm wavelengths) as the principal agent that elicits sunburn, transient inflammation, melanoma and non-melanoma cancer, and premature skin aging. The common form of skin damage described as ‘photoaging’ is caused by repeated sun exposures; it can induce epithelial deoxyribonucleic acid (DNA) damage, and alter the structure and function of dermal connective tissue and extracellular matrix. The present study was aimed to elucidate the effects of a single exposure of different doses of UV-B radiation on the dorsal skin in the hairless mouse. The histopathologic sequels of such exposures were analyzed by using common staining methods, immunohistochemistry, and histomorphometry. Single UV-B exposures seem to elicit dose-graded histological changes some of which have been described in experiments with chronic exposure in this species.     

Effects of chrysotile asbestos on DNA synthesis and growth of human embryonic lung fibroblasts

The in vitro effects of asbestos fibers on thymidine (TdR3H) incorporation and growth of lung fibroblasts have been studied. Incubation of human embryonic lung fibroblasts with UICC Chrysotile B asbestos for 48 hr caused a 3 to 5-fold increase of TdR3H incorporation as compared with control cultures. This increase was dose-dependent with optimal effect obtained with doses as low as 10 micrograms/ml and with cell density of 5 X 10(4) fibroblasts per culture. However, enhanced TdR3H incorporation in treated cells was not correlated with an overall increase of the fibroblast population compared with control cultures as evidenced by cell counts and microscopic examination. Fibroblasts exposed to relatively low concentrations of UICC chrysotile (5-10 micrograms/ml) displayed an initial decrease in cell number compared to controls during the first 24 hr of incubation. At 48 hr however, enhanced TdR3H incorporation occurred with a concomittant increase in cell number. Moreover, continuous exposure of fibroblast cultures to chrysotile (10 micrograms/ml) for a longer period of time led to sustained increase of TdR3H incorporation and resumption of cell proliferation. It is suggested that increased thymidine incorporation is directly related to the effectiveness of asbestos in inhibiting the growth of lung fibroblasts and that measurement of TdR3H incorporation may represent a sensitive means of assessing rapidly the biological activity of asbestos. The possible relevance of this activity to asbestos-induced fibrogenesis and tumorigenesis is also discussed.     

Tetrachlorodibenzo-p-Dioxin (TCDD) Inhibits Differentiation and Increases Apoptotic Cell Death of Precursor T-Cells in the Fetal Mouse Thymus

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) causes thymic atrophy as well as alterations in thymocyte maturity in mice. Multiple mechanisms for thymic hypocellularity have been suggested, and include an increase in thymocyte apoptosis, a maturation arrest of thymocyte development, inhibited thymocyte proliferation, and a diminution of seeding of the thymus by the hematopoietic progenitors in the fetal liver or adult bone marrow. Fetal mice are highly sensitive to hypocellularity induction by TCDD when the chemical is administered during the window of thymic development, between days 10 and 18 of gestation. Treatment of pregnant C57Bl/6 mice in the present experiments with doses of 5 or 10 mu g/kg TCDD by oral gavage on gestation days 14 and 16 severely depressed day 18 thymic cellularity. Histopathologic evaluation of day 18 fetal thymi showed disruption of the normal organ architecture with loss of clear distinction between cortical and medullary regions after TCDD. A decrease in thymocyte density was noted in all regions, and was most dramatic in the cortical zones where pyknotic cells were increased by TCDD treatment. Using day 18 thymocyte suspensions and flow cytometry, the marker 7-AAD showed a decrease in viable thymocytes from TCDD-treated fetal mice, and a concomitant and dose-related increase of thymocytes in early apoptosis. Specifically, relative to control, thymocytes from the 5 and 10 mug/kg TCDD exposure groups displayed 1.9% and 5.3% respective increases in early apoptotic cells. When thymocytes were co-identified by CD4 and CD8 cell surface antigen expression, the enhanced apoptosis occurred in the CD4(+)CD8(+) phenotype with no significant apoptosis seen in the CD4(-)CD8(-), CD4(+)CD8(-), or CD4(-)CD8(+) thymocytes. Given the rapid clearance of apoptotic cells from the thymus, these histopathologic and cytometric data suggest increased thymocyte apoptosis contributes to fetal thymic atrophy after TCDD exposure.     

X-ray potentiation of MuLV infection in vitro.

Physical and chemical agents which cause DNA strand breakage enhance infection by DNA and RNA oncogenic viruses, presumably by facilitating the incorporation of viral genetic information into the host cell genome. X-irradiation has now been shown to potentiate infectious center formation by radiation leukemia virus (RadLV1). The potentiation was dose-dependent with a peak in the range of 300–450 rads. Doses in this range enhanced infectivity by a factor of about 1.5 to 2. X-irradiation also enhanced the infection of cells phenotypically nonpermissive for murine leukemia virus infection, but did not alter the characteristic 2-hit kinetics observed in such cells. Fractionation of the X-ray exposure into two doses separated by varying time intervals showed that the potentiation persisted up to 30 hr postinfection. The initial dose fraction, given at the time of infection, caused partial synchrony of the host cell population. A sharp peak of enhancement occurred when cells received the second dose fraction at a time when they had just completed one round of DNA synthesis and mitosis.     

Low dose exposure to sodium arsenite synergistically interacts with UV radiation to induce mutations and alter DNA repair in human cells

Inorganic arsenic is a known human carcinogen, yet its mechanism of action remains poorly understood. Epidemiological data suggest that arsenic exposure interacts with UV radiation exposure to increase the risk of skin cancer. Studies have suggested that arsenic is able to impair DNA repair enzymes and alter the repair of UV-induced DNA damage. Here we have tested the hypothesis that arsenite [As(III)] and UV interact synergistically to enhance mutagenesis. TK6 human lymphoblastoid cells that are functionally heterozygous at the thymidine kinase (TK) locus were pre-exposed to As(III) alone and in combination with UV. Our data suggest that As(III) is mutagenic only at high doses at the TK locus. As(III) enhanced UV mutagenesis in a more than additive fashion. To investigate the mechanism underlying this synergy we assessed the removal of UV-induced dimers in TK6 cells using the T4 endonuclease-incorporated Comet assay. Pre-treatment with As(III) specifically inhibited the repair of UV-induced pyrimidine dimer-related DNA damage. Taken together, these data suggest that pre-treatment of human cells with arsenic impairs the nucleotide excision repair pathway and leads to enhanced UV mutagenesis.     

Enhanced radiation-induced apoptosis of cancer cell lines after treatment with resveratrol

A search for new agents that can sensitise cancer cells to ionising radiation is of continual interest and mainly due to the use of radiation in cancer therapy. Resveratrol, a powerful antioxidant has been shown to inhibit carcinogenesis in animal models. The purpose of this study was to examine whether resveratrol can sensitise cancer cells to X-irradiation. The human cancer cell lines examined were HELA (cervix carcinoma), K-562 (chronic myeloid leukemia) and IM-9 (multiple myeloma). The assays that were performed following X-irradiation (doses from 0 to 8 Gy) and/or incubation in the presence of resveratrol (concentrations ranging from 0 to 200 microM), were the following: trypan blue exclusion test to determine cell viability, cell morphology after May-Grunwald Giemsa staining, DNA profile analysis by flow cytometry to assess cell cycle distribution and the presence of the sub-G1 peak. The cell lines showed different radiation sensitivity (IM-9, high radiation sensitivity, K-562, intermediate radiation sensitivity and HELA, low radiation sensitivity) as seen by the X-irradiation dose related inhibition of cell growth and induction of apoptosis. The addition of resveratrol alone to the cell cultures induced apoptosis and inhibited cell growth from 50 (IM-9), 100 (EOL-1) or 200 microM (HELA) resveratrol concentrations. Concomitant treatment of the cells with either resveratrol and X-irradiation induced a synergical effect at the highest dose of 200 microM. These results show that resveratrol can act as a potential radiation sensitiser at high concentrations. Further studies need to address the toxicity of resveratrol on normal cells.