Trehalose accumulation and radiation resistance due to prior heat stress in Saccharomyces cerevisiae.

In this study, we examined the accumulation of trehalose, a stress-responsive substance, upon gamma-ray irradiation by evaluating the cause of trehalose accumulation and the development of gamma-ray resistance through intracellular trehalose accumulation. Saccharomyces cerevisiae cells cultured to the logarithmic growth phase were irradiated with gamma rays, and the intracellular trehalose content was measured. However, trehalose was not detectable. The yeast cells with trehalose accumulation caused by pre-treatment at 40 °C were irradiated with gamma rays, and the resistance of these cells to gamma radiation was compared with that of cells without heat treatment. Trehalose accumulation resulted in gamma-ray resistance and suppressed the increase in reactive oxygen species, lipid peroxidation, and DNA double-strand break production in yeast cells. The tests were also performed with a trehalose-6-phosphate-synthase (TPS1)-deficient mutant strain (Δtps1) unable to synthesize trehalose, and the results revealed that TPS1 was involved in protection against oxidative stress.

Hydrogen-rich bath with nano-sized bubbles improves antioxidant capacity based on oxygen radical absorbing and inflammation levels in human serum.

This study compared the effects of hydrogen-water (HW) bath on the oxygen radical absorption-based antioxidant capacity and the inflammatory indicator, C-reactive protein (CRP), in serum between healthy volunteers and inflammatory/collagen disease-patients. The HW bath apparatus supplied nano-bubbles with a diameter of 110 ± 10 nm and 338-682 µg/L of dissolved hydrogen after 120 minutes electrolysis, and nano-bubbles increased to 9.91 × 107/mL along with the increase of correlative dissolved hydrogen. Ten-minute HW bath increased the oxygen radical absorption-based antioxidant capacity to 110.9 ± 9.2% at post-bathing 120 minutes, although unaltered with 10-minute normal water bath at 40°C in healthy subjects. The CRP level was repressed to 70.2 ± 12.1% at 120 minutes after HW bath, although rather increased for normal water bath. In the patients with connective tissue diseases, the CRP level was repressed to 3-24% upon 9 days to 4 months of HW bathing. In another six patients with diverse autoimmune-related diseases, upon daily HW bathing as long as 2-25 months, the pre-bathing CRP level of 5.31 mg/dL decreased to 0.24 mg/dL being within the standard-range, with relief of visible inflammatory symptoms for some cases. Thus, the HW bath with high-density nano-bubbles has beneficial effects on serum antioxidant capacity, inflammation, and the skin appearance. The study was approved by the Committee of Ethics, Japanese Center of Anti-Aging Medical Sciences (Authorization No. H-15-03-2, on January 15, 2019), which was a non-profitable organization officially authenticated by the Hiroshima Prefecture Government of Japan.

Effects of an environmental endocrine disruptor, para-nonylphenol on the cell growth of Euglena gracilis: association with the cellular oxidative stress.

Effects of an environmental endocrine disruptor, para-nonylphenol (NP) on the cell growth of a photosynthetic eukaryotic microorganism, Euglena gracilis were analysed under different cell culture conditions. Although NP did not show significant inhibitory effects on the cell growth of E. gracilis (Z and SM strains) under light culture condition, NP exhibited significant suppressive effects under dark culture condition. Exogenous supplementation with lipophilic antioxidants (α-tocopherol, ß-carotene or 6-O-palmitoyl-ascorbic acid) to E. gracilis caused strong preventive effects against NP-induced cell growth inhibition under dark culture condition, but hydrophilic antioxidants [ascorbic acid, glutathione and epigallocatechin gallate (EGCG)] did not show significant preventive effects. NP caused significant generation of reactive oxygen species (ROS) in E. gracilis under dark culture condition, but E. gracilis under light culture condition did not show significant increase in ROS generation. Supplementation with lipophilic antioxidants to E. gracilis caused significant suppressive effects against NP-induced cellular ROS generation under dark culture condition, but hydrophilic antioxidants did not show significant suppressive effects. Furthermore, the productivities of typical cellular antioxidants (α-tocopherol, ß-carotene and ascorbic acid) in E. gracilis under light culture conditions were much higher than those under dark culture conditions.

Biological effects of low-dose γ-ray irradiation on chromosomes and DNA of Drosophila melanogaster.

While the damage to chromosomes and genes induced by high-dose radiation (HDR) has been well researched in many organisms, the effects of low-dose radiation (LDR), defined as a radiation dose of ≤100 mSv, are still being debated. Recent research has suggested that the biological effects of LDR differ from those observed in HDR. To detect the effect of LDR on genes, we selected a gene of Drosophila melanogaster, known as the multiple wing hair (mwh) gene. The hatched heterozygous larvae with genotype mwh/+ were irradiated by γ-rays of a 60Co source. After eclosion, the wing hairs of the heterozygous flies were observed. The area of only one or two mwh cells (small spot) and that of more than three mwh cells (large spot) were counted. The ratio of the two kinds of spots were compared between groups irradiated by different doses including a non-irradiated control group. For the small spot in females, the eruption frequency increased in the groups irradiated with 20-75 mGy, indicating hypersensitivity (HRS) to LDR, while in the groups irradiated with 200 and 300 mGy, the frequency decreased, indicating induced radioresistance (IRR), while in males, 50 and 100 mGy conferred HRS and 75 and 200 mGy conferred IRR. For the large spot in females, 75 mGy conferred HRS and 100-800 mGy conferred IRR. In conclusion, HRS and IRR to LDR was found in Drosophila wing cells by delimiting the dose of γ-rays finely, except in the male large spot.

Dependencies of hydrogen-water on mineral-based hardness, temperatures and the container materials, and effects of the oral washing and drinking.

Widely distributed electrolytic-generators for hydrogen-water are not fully considered for the dependencies of post-electrolytic values of the dissolved hydrogen concentration (DH) and the oxidation-reduction potential (ORP) on the properties of the pre-electrolytic water. We investigated the dependencies of DH and ORP on mineral-based hardness, temperatures and the container materials, and effects on the oral cavity by oral washing or drinking. Along with an increase in mineral-based water-hardness, DH decreased from 960 to 870 µg/L and the ORP unexpectedly increased from -460 to -320 mV. Purified water of almost zero hardness, however, caused a post-electrolytic DH as low as 80 µg/L and an ORP as high as +20 mV. Post-electrolytic DHs were not significantly changed (780-900 µg/L) upon electrolysis at 1.5-30°C and decreased at 40-50°C. The diffusion of hydrogen from the inside to the outside of the container was extremely small even after 12 hours for an aluminum- or stainless steel-made container, but not for containers made of diverse plastics. The ORP of the intact saliva was +136 mV, and decreased to +90 mV at 20 minutes after 1-minute oral-cramming of hydrogen-water, but returned to +135 mV after 60-minute leaving, showing a transient ORP-decrease in the saliva. Drinking-pause for 4 weeks after drinking hydrogen-water, however, saliva ORP, gradually but not instantly, increased to +60 to +80 mV, but upon drinking-resumption and 2 weeks thereafter, decreased again to -100 to -110 mV, suggesting that several-week hydrogen-water drinking caused a certain decrease in the saliva ORP. Thus, the present study provided the appropriate conditions such as hardness and temperatures for hydrogen-water production by the electrolytic generator, and the container materials suitable for hydrogen-water preservation. Furthermore, we clarified ORP changes of human saliva, being an indicator for human oxidative stress. The study was approved by the Medical Ethics Committee of the NPO (Non-Profitable Organization)-Corporate Japanese Center for Anti-Aging Medical Sciences (approval No. 09S02) on May 2, 2012.

Effects of hydrogen-occluding-silica microparticles on wound repair and cell migratory behavior of normal human esophageal epitheliocytes.

Many conventional studies on molecular hydrogen have not examined cell migration ability and the relationship between apoptosis and the cytoskeleton. Here we investigated the influence of hydrogen-occluding silica microparticles (H2-silica) on cell migration motility and changes of the cytoskeleton (F-actin) in normal human esophageal epithelial cells (HEEpiCs). As the results, cell migration was promoted, and formation of microvilli was activated in the 100 ppm (low concentration) scratched group. After performing a wound healing assay, cells exhibited migration after 48 hours and 72 hours for both 10 ppm and 100 ppm groups, suggesting that the wound-repairing effects could be attributed to the antioxidant ability of H2-silica. In scratched groups, high levels of activated caspase-3 were relatively expressed and presented a tendency to increase the observed Bax/Bcl-2 ratio at more than 300 ppm groups. The above-mentioned results show that H2-silica induced apoptosis in HEEpiCs, especially in the scratched cells. Toxicity may cause an exaggerated apoptosis. Furthermore, since the ratio of fascin/tubulin in the 100, 300, and 600 ppm groups tended to increase in both the scratched and the non-scratched control groups, H2-silica was thought to be able to promote fascin action on normal cells and may be have a proliferative effect.

Associations of the first occurrence of pathogen-specific clinical mastitis with milk yield and milk composition in dairy cows.

The aim of this study was to estimate the associations of the first occurrence of pathogen-specific clinical mastitis (CM) with milk yield and milk composition (somatic cell count (SCC), lactose, fat, protein content in milk and milk urea nitrogen (MUN)). We studied 3149 dairy cows in 31 Hokkaido dairy farms in Japan. Five pathogen groups were studied: Streptococcus spp.; Staphylococcus aureus (S. aureus); coagulase-negative staphylococci (CNS); coliforms; and fungi. Test-day milk data and clinical records were collected from June 2011 until February 2014. Mixed models with an autoregressive correlation structure were fitted to quantify the effects of CM and several other control variables (herd, calving season, parity, week of lactation, and other diseases). Primipara (first lactation) and multipara (second and later lactations) were analysed separately. All pathogens, particularly S. aureus and fungi, were associated with significant milk losses in multipara. In this study, S. aureus and CNS infections were not associated with significant milk loss in primipara. All pathogens, in particular S. aureus and fungi, significantly increased SCC in both parity groups. All pathogens, especially CNS (in primipara) and S. aureus (in multipara), decreased lactose content. All pathogen groups except for fungi were associated with significant changes in fat, protein and MUN. Some pathogens such as Streptococcus spp. and coliforms seemed to be associated with long-term fat, protein and MUN changes. These findings provide estimates that could be used to calculate precise costs of CM, and also provide better indicators of pathogen-specific mastitis.

Electrolytically generated hydrogen warm water cleanses the keratin-plug-clogged hair-pores and promotes the capillary blood-streams, more markedly than normal warm water does.

Biomedical properties of hydrogen water have been extensively investigated, but the effect of hydrogen on good healthy subjects remains unclear. This study was designed to explore the hygiene improvement by electrolytically generated hydrogen warm water (40°C) on capillary blood streams, skin moisture, and keratin plugs in skin pores in normal good healthy subjects with their informed consents. Fingertip-capillary blood stream was estimated after hand-immersing in hydrogen warm water by videography using a CCD-based microscope, and the blood flow levels increased to about 120% versus normal warm water, after 60 minutes of the hand-immersing termination. Skin moisture of subjects was assessed using an electro-conductivity-based skin moisture meter. Immediately after taking a bath filled with hydrogen warm water, the skin moisture increased by 5-10% as compared to before bathing, which was kept on for the 7-day test, but indistinct, because of lower solubility of hydrogen in "warm" water than in room-temperature water. Cleansing of keratin plugs in skin-pores was assessed by stereoscopic microscopy and scanning electron microscopy. After hydrogen warm water bathing, the numbers of cleansed keratin plugs also increased on cheek of subjects 2.30- to 4.47-fold as many as the control for normal warm water. And areas of cleansed keratin plugs in the cheeks increased about 1.3-fold as much as the control. More marked improvements were observed on cheeks than on nostrils. Hydrogen warm water may thoroughly cleanse even keratin-plugs of residual amounts that could not be cleansed by normal warm water, through its permeability into wide-ranged portions of hair-pores, and promote the fingertip blood streams more markedly than merely through warmness due to normal warm water.

Influence of hydrogen-occluding-silica on migration and apoptosis in human esophageal cells in vitro.

In the last decade, many studies have shown that hydrogen gas or hydrogen water can reduce the levels of reactive oxygen species in the living body. Molecular hydrogen has antioxidant and antiapoptotic effects and a preventive effect on oxidative stress-induced cell death. In the present study, we investigated solidified hydrogen-occluding-silica (H2-silica) that can release molecular hydrogen into cell culture medium because the use of hydrogen gas has strict handling limitations in hospital and medical facilities and laboratories, owing to its physicochemical characteristics. Human esophageal squamous cell carcinoma (KYSE-70) cells and normal human esophageal epithelial cells (HEEpiCs) were used to investigate the effects of H2-silica on cell viability and proliferation. Cell migration was examined with wound healing and culture-insert migration assays. The intracellular levels of reactive oxygen species were evaluated with a nitroblue tetrazolium assay. To assess the apoptotic status of the cells, the Bax/Bcl-2 ratio and cleaved caspase-3 were analyzed by western blot. The results showed that KYSE-70 cells and HEEpiCs were generally inhibited by H2-silica administration, and there was a significant proliferation-inhibitory effect in an H2-silica concentration-dependent manner compared with the control group (P < 0.05) in KYSE-70. Apoptosis-inducing effect on KYSE-70 cells was observed in 10, 300, 600, and 1,200 ppm H2-silica, and only 1,200 ppm H2-silica caused a 2.4-fold increase in apoptosis in HEEpiCs compared with the control group as the index of Bax/Bcl-2. H2 silica inhibited cell migration in KYSE-70 cells, and high concentrations had a cytotoxic effect on normal cells. These findings should provide insights into the mechanism of inhibition of H2-silica on human cancer cells in vitro.

Effects of Platinum Nanocolloid in Combination with Gamma Irradiation on Normal Human Esophageal Epithelial Cells.

Our previous study demonstrated that platinum nanocolloid (Pt-nc), combined with lower-dose gamma irradiation at 3, 5, and 7 Gy significantly decreased proliferation and accelerated apoptosis of the human esophageal squamous cell carcinoma-derived cell line KYSE-70. The aim of the present study was to determine, under the same conditions as our previous study where gamma rays combined with Pt-nc were carcinostatic to KYSE-70 cells, if we could induce a radioprotective or the radiation-sensitizing effect on the human normal esophageal epithelial cells (HEEpiC). HEEpiC were treated with various Pt-nc concentrations and then irradiated with various gamma-ray doses. The proliferative status of HEEpiC was evaluated using trypan blue dye-exclusion and WST-8 assays. The cellular and nucleic morphological features were determined using crystal violet and Hoechst 33342 stainings, respectively. The intracellular level of reactive oxygen species (ROS) in HEEpiC was evaluated with a nitro blue tetrazolium (NBT) assay. The apoptotic status was detected with caspase-3, Bax, and Bcl-2 by Western blotting. Either Pt-nc or gamma irradiation could inhibit the growth of HEEpiC; however, their combined use exerted a significant proliferation-inhibitory effect in a Pt-nc dose-dependent manner than gamma irradiation alone. Pt-nc resulted in radiation sensitization rather than radiation protection on HEEpiC in vitro similar to KYSE-70 cells, when Pt-nc was administrated alone or combined with gamma irradiation. Thus, Pt-nc has an inhibitory effect on cell proliferation, a facilitative effect on apoptosis, and a certain degree of toxicity against HEEpiC.

Three-body system metaphor for the two-slit experiment and Escherichia coli lactose-glucose metabolism.

We compare the contextual probabilistic structures of the seminal two-slit experiment (quantum interference experiment), the system of three interacting bodies andEscherichia colilactose-glucose metabolism. We show that they have the same non-Kolmogorov probabilistic structure resulting from multi-contextuality. There are plenty of statistical data with non-Kolmogorov features; in particular, the probabilistic behaviour of neither quantum nor biological systems can be described classically. Biological systems (even cells and proteins) are macroscopic systems and one may try to present a more detailed model of interactions in such systems that lead to quantum-like probabilistic behaviour. The system of interactions between three bodies is one of the simplest metaphoric examples for such interactions. By proceeding further in this way (by playing withn-body systems) we shall be able to find metaphoric mechanical models for complex bio-interactions, e.g. signalling between cells, leading to non-Kolmogorov probabilistic data.

Carcinostatic effects of platinum nanocolloid combined with gamma irradiation on human esophageal squamous cell carcinoma.

AIMS: To explore the carcinostatic effects of platinum nanocolloid (Pt-nc) combined with gamma rays on human esophageal squamous cell carcinoma (ESCC). MAIN METHODS: ESCC-derived KYSE-70 cells were treated with various concentrations of Pt-nc and/or gamma irradiation, and subsequently cultured in phenol red free DMEM with 10% FBS for 48 h. The proliferative status of the KYSE-70 cells was evaluated using trypan blue dye exclusion and WST-8 assays. Cellular and nucleic morphological aspects were evaluated using crystal violet and Hoechst 33342 stainings, respectively. Radiosensitivity was quantified by a cell viability assay, and the activated form of caspase-3, a characteristic apoptosis-related protein, was detected by Western blotting. KEY FINDINGS: Although single treatment with either Pt-nc or gamma irradiation could slightly inhibit the growth of the KYSE-70 cells, their combination exerted remarkable carcinostatic effects in a manner dependent on either Pt-nc concentrations or gamma ray doses, compared with the effect of each treatment alone (p<0.05). By fluorescence micrographic observation, the KYSE-70 cells that were treated with Pt-nc and subsequently irradiated with gamma rays, were shown to undergo distinct apoptotic morphological changes. The carcinostatic effect of gamma rays at 7 Gy without Pt-nc was approximately equal to that when 3-Gy irradiation was combined with 100 ppm Pt-nc or that 5-Gy irradiation was combined with 50 ppm Pt-nc. SIGNIFICANCE: Pt-nc in combination with gamma rays may exert a cooperative effect through platinum- or gamma ray-induced apoptosis resulting in the inhibition of growth of cancer cells, while concurrently enabling the lowering of the radiative dose.

A model of epigenetic evolution based on theory of open quantum systems.

We present a very general model of epigenetic evolution unifying (neo-)Darwinian and (neo-)Lamarckian viewpoints. The evolution is represented in the form of adaptive dynamics given by the quantum(-like) master equation. This equation describes development of the information state of epigenome under the pressure of an environment. We use the formalism of quantum mechanics in the purely operational framework. (Hence, our model has no direct relation to quantum physical processes inside a cell.) Thus our model is about probabilities for observations which can be done on epigenomes and it does not provide a detailed description of cellular processes. Usage of the operational approach provides a possibility to describe by one model all known types of cellular epigenetic inheritance.

Quantum-like model of diauxie in Escherichia coli: operational description of precultivation effect.

In this paper we apply the quantum-like (QL) approach to microbiology to present an operational description of the complex process of diauxie in Escherichia coli. We take as guaranteed that dynamics in cells is adaptive, i.e., it depends crucially on the microbiological context. This very general assumption is sufficient to appeal to quantum and more general QL probabilistic models. The next step is to find the operational representation - by operators in complex Hilbert space (as in quantum physics). To determine QL operators, we used the statistical data from Inada et al. (1996). To improve the QL-representation, we needed better experimental data. Corresponding experiments were recently done by two of the authors and in this paper we use these new data. In these data we found that bio-chemical context of precultivation of populations of E. coli plays a crucial role in E. coli preferences with respect to sugars. Hence, the form of the QL operator representing lactose operon activation also depends crucially on precultivation. One of our results is decomposition of the lactose operon activation operator to extract the factor determined by precultivation. The QL operational approach developed in this paper can be used not only for description of the process of diauxie in E. coli, but also other processes of gene expression. However, new experimental statistical data are demanded.

Quantum-like model for the adaptive dynamics of the genetic regulation of E. coli's metabolism of glucose/lactose.

We developed a quantum-like model describing the gene regulation of glucose/lactose metabolism in a bacterium, Escherichia coli. Our quantum-like model can be considered as a kind of the operational formalism for microbiology and genetics. Instead of trying to describe processes in a cell in the very detail, we propose a formal operator description. Such a description may be very useful in situation in which the detailed description of processes is impossible or extremely complicated. We analyze statistical data obtained from experiments, and we compute the degree of E. coli's preference within adaptive dynamics. It is known that there are several types of E. coli characterized by the metabolic system. We demonstrate that the same type of E. coli can be described by the well determined operators; we find invariant operator quantities characterizing each type. Such invariant quantities can be calculated from the obtained statistical data.

Quantum-like model of brain's functioning: decision making from decoherence.

We present a quantum-like model of decision making in games of the Prisoner's Dilemma type. By this model the brain processes information by using representation of mental states in a complex Hilbert space. Driven by the master equation the mental state of a player, say Alice, approaches an equilibrium point in the space of density matrices (representing mental states). This equilibrium state determines Alice's mixed (i.e., probabilistic) strategy. We use a master equation in which quantum physics describes the process of decoherence as the result of interaction with environment. Thus our model is a model of thinking through decoherence of the initially pure mental state. Decoherence is induced by the interaction with memory and the external mental environment. We study (numerically) the dynamics of quantum entropy of Alice's mental state in the process of decision making. We also consider classical entropy corresponding to Alice's choices. We introduce a measure of Alice's diffidence as the difference between classical and quantum entropies of Alice's mental state. We see that (at least in our model example) diffidence decreases (approaching zero) in the process of decision making. Finally, we discuss the problem of neuronal realization of quantum-like dynamics in the brain; especially roles played by lateral prefrontal cortex or/and orbitofrontal cortex.

Relationship between general anesthesia and memory in Drosophila involving the cAMP/PKA pathways and adhesion-related molecules.

On undergoing an operation under general anesthesia, we tend to lose consciousness, and on recovering from the anesthetic effect, we realize a memory loss during the operation, but do remember the happenings before the operation. It implies that the anesthesia deprivers us of short-term memory without affecting long-term memory. Drosophila melanogaster is known to be an excellent model for genetic studies related to general anesthesia and memory. The various mutants in the genes related to general anesthesia and memory have been found to influence these mechanisms at the molecular level. In Drosophila, learning and memory are classified into four distinct phases: (1) short-term memory (STM), (2) middle-term memory (MTM), (3) longer-lasting anesthesia-resistant memory (ARM), and (4) long-term memory (LTM). On the other hand, based on the genetic studies of the putative target molecules of general anesthetics in model animals, the anesthetic action is classified into five pathways: (1) presynaptic pathway including action potential production, its transmission, and neurotransmitter release; (2) postsynaptic pathway including inhibitory receptors for sleep and pain; (3) memory pathway coupled with cAMP/PKA signaling; (4) adhesion pathway in neuron; and (5) energy production pathway. Memory and adhesion pathways of the anesthetic action are developed in the Drosophila melanogaster model. Many mutants of general anesthesia and those of memory are overlapped suggesting that common molecules and signal pathways are involved in both phenomena. In this review, we will describe the relation between anesthesia and memory, especially highlighting the interaction between the general anesthetics and STM and MTM processes in Drosophila, especially concentrating on the cAMP/PKA signaling and molecular adhesion pathways.

Mapping of putative ether-anesthesia resistance gene using C57BL/6J and MSM/Ms mouse strains.

PURPOSE: We attempted to identify the locations of major mouse genes responsible for sensitivity to diethylether (ether) anesthesia, using microsatellite linkage analyses including Quantitative Trait Locus (QTL) analysis. METHODS: To determine the locations of ether anesthesia resistance genes on chromosomes, an ether anesthesia-resistant mouse strain, C57BL/6J (C57BL), and an ether anesthesia-sensitive mouse strain, MSM/Ms (MSM), were used. The sensitivity of mice to ether anesthesia was determined from the latency time required to lose the righting reflex during exposure to 4% ether vapor in air. The (C57BL x MSM) F(1) mice were found to be resistant to ether, showing that the resistant phenotype is genetically dominant. Twelve resistant and 12 sensitive mice were then selected from the 196 backcrossed F(2) mice (F(1) x MSM) at 11-16 weeks of age. Genomic DNA samples were extracted from the tails for mapping ether anesthesia-related genes using microsatellite linkage analyses. RESULTS: One major putative gene related to resistance to ether anesthesia was restricted in the region 23 to 37 cM from the centromere in chromosome 7 by primary and secondary linkage analyses. The QTL analysis narrowed the position of the gene to 29.0 cM, with a maximum logarithm of odds (LOD) score of 3.03, and it was termed Etan1 ( ether-anesthesia 1). CONCLUSION: Microsatellite linkage analyses, including QTL analysis, determined the location of the ether-resistance gene, Etan1, within a narrow range. Our findings should be helpful for further experiments, such as cloning of the gene governing the sensitivity to ether anesthesia in mice.

Development of a genotoxicity detection system using a biosensor.

The umu-lux test is a genotoxicity test using the two genetically modified S. typhmurium TA1535 strains (TL210 and TL210ctl) transformed with the luxCDABE (luciferase gene and fatty acid reductase genes) of Vibrio fischeri as a reporter gene. The TL210 strain detects genotoxicants and the TL210ctl strain detects cytotoxicants. In order to develop a highly sensitive, simple and rapid genotoxicity detection system, we constructed a biosensor using these immobilized strains. The biosensor consists of two immobilized microbial membranes, a sample vessel and photodetectors, and the genotoxicity detection system consists of the biosensor, an isothermal box, a photodetector and an air pump. The total measurement time for genotoxicants using this detection system is about 4 h. When 2% (v/v) DMSO was used as a control, the TL210 strain was not emitting light while the TL210ctl strain was. When 0.3 mg/l 4NQO was used as a genotoxicant, TL210 strain and TL210ctl strain were both emitting light. When HgCl2 was used as a cytotoxicant, neither the TL210 strain nor the TL210ctl strain were emitting light. Therefore, the false negative prevention function of a biosensor using the TL210ctl strain has been checked. These results show that our proposed system can correctly detect genotoxicants.

Effects of Shaolin Internal Qigong on Physiological Changes / 针灸推拿医学（英文版）

Objective: The physiological changes of the respiro-circulatory functions between skilled subject and unskilled subjects during the practice of Up-right Standing Posture of Shaolin Internal Qigong were compared. Method: The heart rate (HR), maximum oxygen uptake (VO2), respiratory efficiency (RE), and respiratory rate (RR), blood pressure, and lactic acid in the blood were measured.Results: The high correlations between the HR and the VO2 values obtained from the exhaustion test were observed in all subjects. The higher values than at the rest were observed in the average HR and the average VO2 during practicing the Up-right Standing Posture both in the skilled subject and unskilled subjects. However, both HR and VO2 levels were almost constant during the practice in both the skilled subjectand unskilled subjects. The RE values changed in the time course of the practice, whereas the RR values were almost constant during the practice. The average RE showed different patterns between the skilled subject and unskilled subjects, the former increased and the latter decreased their RE levels.In addition, the average values of systolic and diastolic blood pressure of the skilled subject increased each 10 mmHg approximately at pre- and post- Up-right Standing Posture, and the increasing tendency was also recognized in the unskilled subjects. The values of the lactic acid in the blood of the skilled subject increased slightly, whereas the average values of the unskilled subjects increased by 3.4 mmol/1. Conclusion: Shaolin Internal Qigong could improve respiratory efficiency by the maximum isometric muscle contraction while the skilled subject maintained natural breathing. In addition, Shaolin Internal Qigong was considered to influence the reflex system because it inhibited both blood pressure increase and respiratory rate change. Shaolin Internal Qigong fit very well with Tuina doctor training course for promoting the physical ability and manipulation abilities of them.