Low Frequency of MKRN3 Mutations in Central Precocious Puberty Among Korean Girls.

Mutations of MKRN3, the gene encoding makorin RING-finger protein 3, lead to central precocious puberty (CPP). The aim of this study was to investigate mutations of the MKRN3 gene in Korean girls with CPP. Two hundred-sixty Korean girls with idiopathic CPP were included. Auxological and endocrine parameters were measured, and the entire MKRN3 gene was directly sequenced. MKRN3 gene analysis revealed one novel nonsense mutation (p.Gln281 *) and 6 missense variants (p.Ile100Phe, p.Gly196Val, p.Ile204Thr, p.Gln226Pro, p.Lys233Asn, and p.Ser396Arg). The novel nonsense mutation (p.Gln281 *) was a heterozygous C>T nucleotide change (c.841C>T) predicted to result in a truncated protein due to a premature stop codon in the MKRN3 gene. The nonsense mutation (p.Gln281 *) was only identified in one of the girls and her younger brother. Compared to previous reports on MKRN3 mutations in familial and sporadic cases of CPP, the present study reveals a relatively low number of MKRN 3 mutations in Korean girls with CPP. Larger samples of children with CPP and MKRN3 mutations are necessary in order to clarify whether the clinical course of puberty may differ as compared to idiopathic CPP.

Bystander effect between zebrafish embryos in vivo induced by high-dose X-rays.

We employed embryos of the zebrafish, Danio rerio, for our studies on the in vivo bystander effect between embryos irradiated with high-dose X-rays and naive unirradiated embryos. The effects on the naive whole embryos were studied through quantification of apoptotic signals at 25 h post fertilization (hpf) through the terminal dUTP transferase-mediated nick end-labeling (TUNEL) assay followed by counting the stained cells under a microscope. We report data showing that embryos at 5 hpf subjected to a 4-Gy X-ray irradiation could release a stress signal into the medium, which could induce a bystander effect in partnered naive embryos sharing the same medium. We further demonstrated that this bystander effect (induced through partnering) could be successfully suppressed through the addition of the nitric oxide (NO) scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) into the medium but not through the addition of the CO liberator tricarbonylchloro(glycinato)ruthenium(II) (CORM-3). This shows that NO was involved in the bystander response between zebrafish embryos induced through X-ray irradiation. We also report data showing that the bystander effect could be successfully induced in naive embryos by introducing them into the irradiated embryo conditioned medium (IECM) alone, i.e., without partnering with the irradiated embryos. The IECM was harvested from the medium that had conditioned the zebrafish embryos irradiated at 5 hpf with 4-Gy X-ray until the irradiated embryos developed into 29 hpf. NO released from the irradiated embryos was unlikely to be involved in the bystander effect induced through the IECM because of the short life of NO. We further revealed that this bystander effect (induced through IECM) was rapidly abolished through diluting the IECM by a factor of 2× or greater, which agreed with the proposal that the bystander effect was an on/off response with a threshold.

The threshold number of protons to induce an adaptive response in zebrafish embryos.

In this study, microbeam protons were used to provide the priming dose to induce an in vivo radioadaptive response (RAR) in the embryos of zebrafish, Danio rerio, against subsequent challenging doses provided by x-ray photons. The microbeam irradiation system (Single-Particle Irradiation System to Cell, acronym SPICE) at the National Institute of Radiological Sciences (NIRS), Japan, was employed. The embryos were dechorionated at 4 h post fertilisation (hpf) and irradiated at 5 hpf by microbeam protons. For each embryo, one irradiation point was chosen, to which 5, 10, 20, 30, 40, 50, 100, 200, 300 and 500 protons each with an energy of 3.4 MeV were delivered. The embryos were returned to the incubator until 10 hpf to further receive the challenging exposure, which was achieved using 2 Gy of x-ray irradiation, and then again returned to the incubator until 24 hpf for analyses. The levels of apoptosis in zebrafish embryos at 25 hpf were quantified through terminal dUTP transferase-mediated nick end-labelling (TUNEL) assay. The results revealed that at least 200 protons (with average radiation doses of about 300 and 650 mGy absorbed by an irradiated epithelial and deep cell, respectively) would be required to induce RAR in the zebrafish embryos in vivo. Our previous investigation showed that 5 protons delivered at 10 points on an embryo would already be sufficient to induce RAR in the zebrafish embryos. The difference was explained in terms of the radiation-induced bystander effect as well as the rescue effect.

Adaptive response to ionising radiation induced by cadmium in zebrafish embryos.

An adaptive response is a biological response where the exposure of cells or animals to a low priming exposure induces mechanisms that protect the cells or animals against the detrimental effects of a subsequent larger challenging exposure. In realistic environmental situations, living organisms can be exposed to a mixture of stressors, and the resultant effects due to such exposures are referred to as multiple stressor effects. In the present work we demonstrated, via quantification of apoptosis in the embryos, that embryos of the zebrafish (Danio rerio) subjected to a priming exposure provided by one environmental stressor (cadmium in micromolar concentrations) could undergo an adaptive response against a subsequent challenging exposure provided by another environmental stressor (alpha particles). We concluded that zebrafish embryos treated with 1 to 10 µM Cd at 5 h postfertilisation (hpf) for both 1 and 5 h could undergo an adaptive response against subsequent ~4.4 mGy alpha-particle irradiation at 10 hpf, which could be interpreted as an antagonistic multiple stressor effect between Cd and ionising radiation. The zebrafish has become a popular vertebrate model for studying the in vivo response to ionising radiation. As such, our results suggested that multiple stressor effects should be carefully considered for human radiation risk assessment since the risk may be perturbed by another environmental stressor such as a heavy metal.

The multiple stressor effect in zebrafish embryos from simultaneous exposure to ionising radiation and cadmium.

Living organisms are exposed to a mixture of environmental stressors, and the resultant effects are referred to as multiple stressor effects. In the present work, we studied the multiple stressor effect in embryos of the zebrafish (Danio rerio) from simultaneous exposure to ionising radiation (alpha particles) and cadmium through quantification of apoptotic signals at 24 h postfertilisation (hpf) revealed by vital dye acridine orange staining. For each set of experiments, 32-40 dechorionated embryos were deployed, which were divided into four groups each having 8-10 embryos. The four groups of embryos were referred to as (1) the control group (C), which received no further treatments after dechorionation; (2) the Cd-dosed and irradiated group (CdIr), which was exposed to 100 µM Cd from 5 to 24 hpf, and also received about 4.4 mGy from alpha particles at 5 hpf; (3) the irradiated group (Ir), which received about 4.4 mGy from alpha particles at 5 hpf; and (4) the Cd-dosed group (Cd), which was exposed to 100 µM Cd from 5 to 24 hpf. In general, the CdIr, Ir and Cd groups had more apoptotic signals than the C group. Within the 12 sets of experimental results, two showed significant synergistic effects, one showed a weakly synergistic effect and nine showed additive effects. The multiple stressor effect of 100 µM Cd with ~4.4 mGy alpha-particle radiation resulted in an additive or synergistic effect, but no antagonistic effect. The failure to identify significant synergistic effects for some sets of data, and thus their subsequent classification as additive effects, might be a result of the relatively small magnitude of the synergistic effects. The results showed that the radiation risk could be perturbed by another environmental stressor such as a heavy metal, and as such a realistic human radiation risk assessment should in general take into account the multiple stressor effects.

Hormetic effect induced by alpha-particle-induced stress communicated in vivo between zebrafish embryos.

We report data showing that embryos of the zebrafish, Danio rerio, at 1.5 h post fertilization (hpf) subjected to a low-dose alpha-particle irradiation can release a stress signal into the water, which can be communicated to unirradiated bystander zebrafish embryos sharing the same water medium to induce a hormetic effect in the bystander embryos. Hormetic responses are characterized as biphasic dose-response relationships exhibiting a low-dose stimulation and a high-dose inhibition. The effects on the whole embryos were studied through quantification of apoptotic signals at 24 hpf through staining with the vital dye acridine orange, followed by counting the stained cells under a microscope. The results show that, for low alpha-particle dose, the number of apoptotic signals decreases in the irradiated embryos and also in the unirradiated bystander embryos having partnered with the irradiated embryos. These suggested that alpha-particle-irradiated zebrafish embryos could release a stress signal into the water, which could be communicated to unirradiated bystander zebrafish embryos sharing the same water medium to induce a hormetic effect in the bystander embryos.

α-Particle irradiated zebrafish embryos rescued by bystander unirradiated zebrafish embryos.

We report data demonstrating that zebrafish embryos irradiated by α-particles can release a stress signal into the water, which can be communicated to the unirradiated zebrafish embryos sharing the same water medium, and then these unirradiated zebrafish embryos can release a feedback stress signal back to the irradiated embryos. The effects of radiation on the whole embryos were studied through quantification of apoptotic signals at 24 h post fertilization through staining with the vital dye acridine orange, followed by counting the stained cells under a microscope. We refer to this phenomenon as the "rescue effect", where the unirradiated embryos successfully helped the irradiated embryos mitigate the radiation induced DNA damages. The results showed that the number of apoptotic signals in the irradiated embryos was smaller when they were partnered with bystander unirradiated embryos in the same medium. The results also showed significantly fewer apoptotic signals in the irradiated embryos when the population of bystander embryos increased from 10 to 30, while keeping the population of irradiated embryos at 10. These data suggest that the stress communicated between the unirradiated zebrafish embryos and the irradiated embryos sharing the same medium will help "rescue" the irradiated embryos, and that the strength of the rescue effect depends on the number of rescuing bystander unirradiated embryos.

Radioadaptive response induced by alpha-particle-induced stress communicated in vivo between zebrafish embryos.

We report data demonstrating that zebrafish embryos irradiated by alpha particles can release a stress signal into the water, which can be communicated to the unirradiated zebrafish embryos sharing the same water medium and thereby inducing a radioadaptive response in these unirradiated zebrafish embryos. The effects of radiation on the whole embryos were studied through quantification of apoptotic signals at 24 h post fertilization through staining with the vital dye acridine orange, followed by counting the stained cells under a microscope. In these experiments, dechorionated embryos were irradiated and then partnered with two other groups of unirradiated embryos, namely the bystander group (no more further treatments) and adaptive group (subjected to a further challenging dose) of embryos. The adaptive group of embryos were then separately further irradiated with a challenging dose. The results show that the number of apoptotic signals for the adaptive group is smaller than that for the corresponding control group, while that for the bystander group is larger than that for the corresponding control group. These suggest that the stress communicated in vivo between the irradiated zebrafish embryos and those unirradiated embryos sharing the same medium will induce radioadaptive response in the unirradiated embryos.

Observer agreement of the Manchester Triage System and the Emergency Severity Index: a simulation study.

OBJECTIVES: To compare inter and intra-observer agreement of the Manchester Triage System (MTS) and the Emergency Severity Index (ESI). METHODS: 50 representative emergency department (ED) scenarios derived from actual cases were presented to 18 ED nurses from three different hospitals. Eight of them were familiar with MTS, six with ESI and four were not familiar but trained in both systems. They independently assigned triage scores to each scenario according to the triage system(s) they were familiar with. After 4-6 weeks the same nurses again judged the scenarios in a different order. Unanimity in judgement and unweighted and quadratic-weighted kappas were calculated. RESULTS: Unanimity in judgement for MTS was 90% and for ESI 73%. One-level disagreement was found in 8% and 23% of the cases, respectively. Interobserver unweighted kappas were 0.76 (95% CI 0.68 to 0.83) for MTS and 0.46 (95% CI 0.37 to 0.55) for ESI. Quadratic-weighted kappas were 0.82 (95% CI 0.74 to 0.89) and 0.73 (95% CI 0.64 to 0.83), respectively. At 4-6 weeks, one-level intra-observer disagreements were 10% and 22% and 2-level disagreement 1% and 2%, respectively. Intra-observer unweighted kappas were 0.84 (95% CI 0.73 to 0.94) for MTS and 0.65 (95% CI 0.59 to 0.72) for ESI. CONCLUSION: Using paper-based clinical scenarios, MTS was found to have a greater inter and intra-observer agreement than ESI.

Using formal concept analysis for microarray data comparison.

Microarray technologies, which can measure tens of thousands of gene expression values simultaneously in a single experiment, have become a common research method for biomedical researchers. Computational tools to analyze microarray data for biological discovery are needed. In this paper, we investigate the feasibility of using formal concept analysis (FCA) as a tool for microarray data analysis. The method of FCA builds a (concept) lattice from the experimental data together with additional biological information. For microarray data, each vertex of the lattice corresponds to a subset of genes that are grouped together according to their expression values and some biological information related to gene function. The lattice structure of these gene sets might reflect biological relationships in the dataset. Similarities and differences between experiments can then be investigated by comparing their corresponding lattices according to various graph measures. We apply our method to microarray data derived from influenza-infected mouse lung tissue and healthy controls. Our preliminary results show the promise of our method as a tool for microarray data analysis.

Embryos of the zebrafish Danio rerio in studies of non-targeted effects of ionizing radiation.

The use of embryos of the zebrafish Danio rerio as an in vivo tumor model for studying non-targeted effects of ionizing radiation was reviewed. The zebrafish embryo is an animal model, which enables convenient studies on non-targeted effects of both high-linear-energy-transfer (LET) and low-LET radiation by making use of both broad-beam and microbeam radiation. Zebrafish is also a convenient embryo model for studying radiobiological effects of ionizing radiation on tumors. The embryonic origin of tumors has been gaining ground in the past decades, and efforts to fight cancer from the perspective of developmental biology are underway. Evidence for the involvement of radiation-induced genomic instability (RIGI) and the radiation-induced bystander effect (RIBE) in zebrafish embryos were subsequently given. The results of RIGI were obtained for the irradiation of all two-cell stage cells, as well as 1.5 hpf zebrafish embryos by microbeam protons and broad-beam alpha particles, respectively. In contrast, the RIBE was observed through the radioadaptive response (RAR), which was developed against a subsequent challenging dose that was applied at 10 hpf when <0.2% and <0.3% of the cells of 5 hpf zebrafish embryos were exposed to a priming dose, which was provided by microbeam protons and broad-beam alpha particles, respectively. Finally, a perspective on the field, the need for future studies and the significance of such studies were discussed.

Genetic association analysis of serotonin system genes in bipolar affective disorder.

OBJECTIVE: This study examined the putative role of serotonin genes in the etiology of bipolar affective disorder. METHOD: Genetic association analysis was performed for individuals with bipolar affective disorder and unaffected subjects closely matched in age, sex, and ethnic background (N=103 in each group). The allele and genotype frequencies of polymorphisms at the genes for serotonin receptors HTR1A, HTR1Dalpha, HTR1Dbeta, HTR2A, HTR2C, HTR7, tryptophan hydroxylase (TPH), and the serotonin transporter (hSERT) were compared in the two groups of subjects. RESULTS: Statistically significant positive associations were found for HTR2A and hSERT polymorphisms. However, results from an independent replication group of over 100 patients with bipolar affective disorder and their matched comparison subjects failed to confirm these associations. CONCLUSIONS: These results suggest that the serotonin genes studied are not associated with bipolar affective disorder, although transmission disequilibrium studies are required in order to confirm this conclusion.

Occurrence sampling in a residence hall foodservice: entrée production times.

Occurrence sampling was used in the main production area of a university residence hall foodservice to measure and analyze work functions involved in entrée production and to determine time requirements for entrée categories. Sixteen entrées were investigated: seven single-item, six combination, and three roast. Data collection took place in a period of 5 days over a 2-week span. A total of 3,891 observations was recorded during 297 labor hours. Results of the study indicated 59.2% of the total labor time was spent in direct work, 23.3% in indirect work, and 17.5% in delays. The preparation work function represented the highest proportion of labor time for both single-item and roast entrées, but prepreparation consumed the greatest amount of time for combination entrées. Mean labor minutes per serving for the 16 entrées were 0.9, with a range of 0.4 to 3.5. Mean labor minutes per serving for combination entrées were 1.4, which was the highest among the three groups; range was from 0.4 to 3.5 minutes per serving. For single item entrées, mean minutes per serving were 0.6, with a range of 0.4 to 0.8. Roast entrées, had a mean time requirement per serving of 0.5 minute, with a range of 0.4 to 0.6.

Assessment of the protein quality of nine northern adapted yellow and brown seed coated soybean cultivars by amino acid analysis.

Accurate and detailed amino acid determinations were carried out on nine northern adapted soybean cultivars to ascertain whether their amino acid profiles could be used as potentially useful indices for assessing their protein quality. The cultivars were Maple Amber, Maple Donovan, Maple Glen, Maple Isle, Maple Presto, Maple Ridge, and three brown seed coat near-isogenic lines, Maple Presto Brown, Maple Ridge Brown, and Maple Arrow Brown. Their total protein and amino acid composition were compared with those of an established cultivar, Maple Arrow. Mean protein values for the new cultivars ranged from 30.1 to 33.1% compared to Maple Arrow, which was 33.2%. The total nitrogen content was also variable among these cultivars, ranging from 5.0 to 5.4%. All nine Maple series soybean cultivars were higher in their essential amino acid (EAA) content, that is, EAA(9) = 45.2-46.5%, than the FAO/WHO reference protein pattern value of EAA(9) = 33.9%, for a 2-5-year-old child. Each of the nine new soybean cultivars was limited only in methionine and to a lesser extent in valine and isoleucine and had a protein digestibility corrected amino acid score of 91% for all cultivars, compared to the value of egg protein (97%). These results suggest that the most accurate evaluation of protein quality in soybeans, and possibly other legumes and cereals, is by the protein digestibility-corrected amino acid score.

Assessment of the protein quality of 15 new northern adapted cultivars of quality protein maize using amino acid analysis.

Amino acid determinations were carried out on 15 new northern adapted cultivars of quality protein maize (QPM) containing opaque-2 modifier genes to ascertain whether their amino acid scoring patterns could be used to select high-lysine QPM genotypes and to assess their protein quality. Total protein in these cultivars ranged from 8.0 to 10.2% compared to two commercial maize varieties, Dekalb DK435 (7.9%) and Pioneer 3925 (10.3%). Four of these QPM genotypes, QPM-C26, QPM-C21, QPM-C79, and QPM-C59, contained high levels of lysine (4.43-4.58 g of lysine/100 g of protein), whereas the remaining varied from 3.43 to 4.21 g of lysine/100 g of protein, compared to Dekalb DK435 and Pioneer 3925, which contained 2.9 and 3. 1 g of lysine/100 g of protein, respectively. Although lysine is the first limiting amino acid in QPM inbreds, the high-lysine QPM genotypes may supply approximately 70.2-72.6% of human protein requirements, compared to 46.2% for Dekalb DK435 and 50.1% for Pioneer 3925, 55-63% for oats, and 59-60.3% for barley. Northern adapted QPM genotypes may have the potential to increase their lysine content even further, either by an increase in specific high-lysine-containing nonzein proteins, such as the synthesis of factor EF-1a, or by a further reduction in the 19 and 22 kDa alpha-zein in the endosperm or both. This knowledge could assist maize breeders in the selection of new high-performance QPM genotypes with improved protein quality and quantity.

The statistical variability of blood alcohol concentration measurements in drink-driving cases.

Like many other places in the world, Hong Kong has drink-driving legislation which prohibits a driver from having in his blood alcohol exceeding a prescribed limit while in control of a motor vehicle. The accuracy of measuring this alcohol concentration is obviously of prime concern as an erroneous result can avert the administration of justice. The common practice is to deduct all errors from the measured value and compare the deducted value with the prescribed limit, so that the benefit of all errors of the measurement is given to the driver. It is therefore important for any laboratory responsible for measuring blood alcohol concentrations to identify and quantify all errors associated with the measurement. The present study examined 900 blood alcohol determinations carried out by the Hong Kong Government Laboratory (HKGL) on cases of suspected drink driving. The determinations were performed by 5 different analysts with two different sets of instruments during 1995-1997. Statistical analysis indicated that the instruments had no bearing on the random error or variability and that even though analyst was a significant factor on variability, the deviation from the mean so caused was only 0.3% and of no practical significance. When the systematic error introduced by the tolerance limits of the certified alcohol standards (purchased from the Laboratory of Government Chemists, UK) was taken into account, the total uncertainty (random plus systematic errors) of an alcohol determination at 99.5% confidence level was found to be 4%. It is recommended that laboratories engaged in blood alcohol determination should adopt similar statistical treatment of their analytical results to find out the error and to ensure that the results are independent of analyst and instrument used.

Some properties of the signals involved in unirradiated zebrafish embryos rescuing α-particle irradiated zebrafish embryos.

PURPOSE: The in vivo radiation-induced bystander effect (RIBE) and radiation-induced rescue effect (RIRE) induced between embryos of the zebrafish (Danio rerio) by alpha-particle irradiation were studied through the number of apoptotic signals revealed at 24 h post fertilization (hpf) through vital dye acridine orange staining. MATERIALS AND METHODS: RIBE and RIRE were verified through the significant increase and decrease in apoptotic signals in the partnered bystander and irradiated embryos, respectively. RESULTS: The medium transfer experiment where irradiated zebrafish embryos were rescued through immersion in the medium previously conditioned by a larger number of irradiated zebrafish embryos showed (a) the involvement of a released stress signal in the induction of RIRE, and (b) RIBE and RIRE signals had the same function. With the help of 500 µM of the specific nitric oxide (NO) scavenger cPTIO (2-(4carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide), NO was confirmed as an essential signaling molecule for inducing both the RIBE and RIRE. On the other hand, the treatment with 20 µM of the carbon monoxide (CO) releasing chemical CORM-3 (tricarbonylchloro(glycinato)ruthenium (II)) suppressed the manifestations of RIBE but did not suppress RIRE. CONCLUSIONS: In conclusion, unirradiated zebrafish embryos need NO but not NO-induced damages to rescue α-particle irradiated zebrafish embryos.

Development and evaluation of a multiplex PCR for eight plasmid-mediated quinolone-resistance determinants.

The objective of this study was to develop and validate an expanded multiplex PCR assay for the simultaneous detection of eight plasmid-mediated quinolone-resistance determinants in Enterobacteriaceae. Primers were designed to amplify conserved fragments of qnrABCDS, qepA, oqxAB and aac(6')-Ib-cr genes and were optimized in uniplex and multiplex PCR assays with control template DNA. The assay was used to determine the prevalence of plasmid-mediated quinolone resistance (PMQR) genes in 174 ciprofloxacin-resistant and 43 ciprofloxacin-susceptible extraintestinal pathogenic Escherichia coli isolates. Each resistance gene could be detected alone and in combination. PMQR determinants were detected in 65 ciprofloxacin-resistant isolates (37 %) and one ciprofloxacin-susceptible isolate (2 %). Prevalences of the identified determinants were: aac(6')-Ib-cr, 34.5 %; qnrS, 1.1 %; qepA, 1.1 %; and oqxAB, 0.6 %. In conclusion, we developed an eight-target multiplex PCR for the accurate detection of PMQR genes and confirmed that PMQR prevalence remains low among human Escherichia coli clinical isolates in the UK.

A diamond nanocone array for improved osteoblastic differentiation.

Efficient delivery of biomolecules to cells is of great importance in biology and medicine. To achieve this, we designed a novel type of densely packed diamond nanocone array to conveniently transport molecules to the cytoplasm of a great number of cells. The nanocone array was fabricated by depositing a thin layer of diamond film on a silicon substrate followed by bias-assisted reactive ion etching. The height of the diamond nanocones varied from 200 nm to 1 µm with tip radii of approximately 10 nm. Our fluorescein and propidium iodide staining results clearly demonstrated that dramatically enhanced delivery of fluorescein into cells was realized without leading to noticeable cell death with the aid of nanocone treatment. As a test case of the drug delivery application of the device, MC-3T3 cells in differentiation medium were applied to the nanocone array for enhanced intracellular delivery of the medium. This was confirmed by the fact that nanocone treated cells experienced much higher differentiation ability at an early stage in comparison with untreated cells. Overall, the results indicate that the diamond nanocone array provides a very simple but yet very effective approach to achieve delivery of molecules to a large number of cells.

Alpha radiation exposure decreases apoptotic cells in zebrafish embryos subsequently exposed to the chemical stressor, Cd.

The aim of this study was to demonstrate that zebrafish embryos subjected to a priming exposure provided by one environmental stressor (low-dose alpha particles) can induce an adaptive response against a subsequent challenging exposure provided by another environmental stressor (heavy metal Cd). The effect thus identified would be an antagonistic multiple stressor effect. The effects of alpha particle radiation and/or Cd on whole embryos were studied through quantification of apoptotic signals at 24 h post-fertilization (hpf). Embryos were stained with the vital dye acridine orange, followed by counting the stained cells. For each set of experiments, 30 dechorionated embryos were divided into three groups, each having ten embryos. The three groups of embryos were referred to as (A) the control group, which received no more further treatments after dechorionation, (B) Cd-treated group, which did not receive any priming exposure and would receive a challenging exposure at 10 hpf and (C) (alpha + Cd)-treated group, which would receive both priming and challenging exposures. We defined the normalized net number of apoptotic signals in the (alpha + Cd)-treated group as N (C) * = [(apoptotic signals for (alpha + Cd)-treated group - average apoptotic signals for the corresponding control group)/average apoptotic signals for the corresponding control group] and that in the Cd-treated group as N (B)* = [(apoptotic signals for Cd-treated group - average apoptotic signals for the corresponding control group)/ average apoptotic signals for the corresponding control group]. By using the non-parametric Mann-Whitney U statistic, we were able to show that N (C) * was significantly smaller than N (B) *(p = 0.006). These demonstrated an antagonistic multiple stressor effect between ionizing radiation and Cd through the induction of an adaptive response by the ionizing radiation against subsequent exposures to Cd.