Specific photoisomerization of retinal in squid rhodopsin and metarhodopsin

The composition of retinal isomers in the photosteady-state mixtures formed from squid rhodopsin and metarhodopsin was determined by high-pressure liquid chromatography. A large amount of 9-cis-retinal was obtained at liquid N₂ temperature when rhodopsin was irradiated with orange light, but only small quantities of 9-cis-retinal were obtained at 15°C. Scarcely any 9-cis-retinal was produced from metarhodopsin by irradiation at liquid N₂ temperature. A large quantity of 7-cis-retinal was found in the photoproduct of rhodopsin irradiated at solid carbon dioxide temperature, but not at 15°C and liquid N₂ temperature. 7-cis-Retinal was not produced from metarhodopsin at any temperatures. These results indicate that the photoisomerization of retinal is regulated by the structure of the retinal-binding site of this protein. The formation of 9-cis- and 7-cis-retinals is forbidden in the metarhodopsin protein.     

Temperature Effects on Bacterial Phytochrome

Bacteriophytochromes (BphPs) are light-sensing regulatory proteins encoded in photosynthetic and non-photosynthetic bacteria. This protein class incorporate bilin as their chromophore, with majority of them bearing a light- regulated His kinase or His kinase related module in the C-terminal. We studied the His kinase actives in the temperature range of 5°C to 40°C on two BphPs, Agp1 from Agrobacterium tumefaciens and Cph1 from cyanobacterium Synechocystis PCC 6803. As reported, the phosphorylation activities of the far red (FR) irradiated form of the holoprotein is stronger than that of the red (R) irradiated form in both phytochromes. We observed for the apoprotein and FR irradiated holoprotein of Agp1 an increase in the phosphorylation activities from 5°C to 25°C and a decrease from 25°C to 40°C. At 5°C the activities of the apoprotein were significantly lower than those of the FR irradiated holoprotein, which was opposite at 40°C. A similar temperature pattern was observed for Cph1, but the maximum of the apoprotein was at 20°C while the maximum of the FR irradiated holoprotein was at 10°C. At 40°C, prolonged R irradiation leads to an irreversible bleaching of Cph1, an effect which depends on the C-terminal His kinase module. A more prominent and reversible temperature effect on spectral properties of Agp1, mediated by the His kinase, has been reported before. His kinases in phytochromes could therefore share similar temperature characteristics. We also found that phytochrome B mutants of Arabidopsis have reduced hypocotyl growth at 37°C in darkness, suggesting that this phytochrome senses the temperature or mediates signal transduction of temperature effects.     

The effect of hematoporphyrin and light on human platelets. I. Morphologic, functional, and biochemical changes

The morphologic, functional, and biochemical changes produced by hematoporphyrin and light in human platelets have been characterized. by phase microscopy the cells appeared swollen and resembled signet rings; by electron microscopy they showed considerable loss of cytoplasm and their contour was smoother than normal. irradiated platelets were not aggregable by thrombin and calcium chloride, although they contained clottable protein, and were incapable of supporting clot retraction. a linear relationship was demonstrated between the per cent depletion of serotonin from irradiated platelets and the log dose of hematoporphyrin. the depletion of serotonin from these platelets was related lineraly to the log of time of exposure to light during the initial six minutes of exposure; but thereafter continued at a constant rate. the temperature of incubation influenced directly the rate of depletion of serotonin from irradiated platelets but did not influence the movement of serotonin into these platelets. atp was diminished considerably in irradiated platelets. these changes are attributable to damage to the membrane of the platelet by hematoporphyrin and light. These studies provide additional information about the blood platelet in terms of its response to photodynamic action.     

Inhibition of mitosis in pisum root meristems by continuous gamma radiation: the influence of temperature on the synthesis of DNA, RNA and protein during inhibition

Tritiated precursors of DNA, RNA and protein were used to measure synthesis at 10 and 20C in root meristem cells of Pisum after they were mitotically arrested by continuous irradiation with gamma rays. The experiments were designed to determine if the arrested cells accumulated in a certain part of interphase, to determine the effect on DNA, RNA and protein synthesis, to find out if the effects were temperature dependent, and finally to reveal possible relationships between growth inhibition and altered synthesis. The results showed that the incorporation of DNA and RNA precursors was impaired by irradiation and that decreased temperature further increased radiation impairment of DNA synthesis. Protein synthesis on the other hand was not impaired by irradiation at either temperature. Irradiation at 20C reduced the number of DNA-synthesizing cells; at 10C this number was reduced to near zero. Although irradiated cells synthesizing RNA showed a reduction in grain counts when compared to the controls, they still retained the ability to incorporate tritiated uridine at 10C. It was hypothesized that the combination of reduced DNA and RNA synthesis and unaffected protein synthesis resulted in precocious maturation of the arrested meristem cells. Growth which occurred in the absence of cell division was attributed to meristematic cells which precociously matured and cells which were in the region of elongation.     

Stimulation of alkalothermophilic Aspergillus terreusxylanase by low-intensity laser radiation

In this study, Aspergillus terreus was irradiated by a 7.3 mW He–Ne laser in the presence of crystal violet, toluidine blue O and hematoporphyrin as photosensitizers. Xylanases recovered from non-irradiated and irradiated fungi were purified and characterized. The maximum production of xylanase (42.2 U/ml) was obtained after 5 min of laser irradiation in the absence of the photosensitizer. The irradiation of the sensitized fungus diminished the production of xylanase. On purification using G-100, the specific activity of xylanase recovered from the irradiated fungus was 292 U/mg protein representing a 37-fold purification over the crude extract compared with 95.6 U/mg protein representing the 12.8-fold for the enzyme recovered from the non-irradiated fungus. The enzyme recovered from the irradiated fungus had lower molecular weight as compared with that recovered from the non-irradiated one. Characterization of the purified enzymes revealed that the enzyme recovered from the irradiated fungus was more thermostable and had a wider range of optimum reaction temperature (60–70°C) and pH (4.0–12.0), compared to the non-irradiated one.     

Effect of temperature on the electron paramagnetic resonance spectrum of irradiated alanine.

Polycrystalline samples of the amino acid L-alpha-alanine have been irradiated with X rays at both room temperature and higher temperatures. The electron paramagnetic resonance (EPR) spectra of alanine powder irradiated at room temperature are dominated by the well-known room-temperature-stable alanine radical CH3C*HCOOH. Upon heating of room-temperature-irradiated alanine powder, a strong decay of the signal was observed, and the features of the spectrum recently ascribed to a second stable radical in alanine irradiated at room temperature become more pronounced, providing an experimental isolation of this second alanine radical. In combination with the high-temperature experiments, a multivariate statistical decomposition method, maximum likelihood common factor analysis, was used to determine the number of components in irradiated alanine powder which behave differently as a function of temperature. The EPR components found in the present study are compared with simulations using earlier EPR and ENDOR single-crystal data.     

The Effect of Freezing on the Radiation Sensitivity of Bacterial Spores

S ummary : Bacillus pumilus spores, irradiated under aerobic conditions, were inactivated exponentially at the same rate whether they were at room temperature (10–13°) in phosphate buffer or at -79° in phosphate buffer or in heart infusion broth.
Clostridium welchii spores were irradiated in Robertson's cooked meat medium under anaerobic conditions. With unheated spores, and those subjected to a heat shock before irradiation, the inactivation rate was the same at room temperature and -79°. The same applied to spores heat shocked after irradiation for doses up to 450 Krads, but above this dose level the spores irradiated frozen were more sensitive.
The effect of the heat shock, whether applied before or after irradiation, was to increase the number of survivors, and the proportionate increase appeared to vary with dose.     

造血器官机能障碍后家蚕血淋巴中蛋白质成分的变化

为了研究家蚕Bombyx mori造血器官机能障碍后其血淋巴中蛋白质成分的变化,利用重离子射线局部照射家蚕幼虫的造血器官,检测了照射后家蚕血淋巴中的蛋白质成分及注射大肠杆菌后在体内诱导出现的应急蛋白量的变化。结果表明,照射蚕血淋巴中的蛋白质含量与对照蚕之间没有明显的差异。但在成分分析时发现,5龄起蚕血淋巴中70 kD附近的3条蛋白质谱带比对照蚕的浓度要高,随着个体的发育两者的浓度都上升;5龄后期则相反,对照蚕的浓度比照射蚕高;脂肪体中贮藏蛋白质的含量具有相似的变化趋势。用家蚕贮藏蛋白质SP-1及SP-2的抗血清进行免疫印迹反应的结果显示：70 kD附近的3条蛋白质谱带的最上面的一条为贮藏蛋白质SP-1,下面的二条为贮藏蛋白质SP-2;同时照射蚕血淋巴中分子量约为24 kD的蛋白质成分也发生变化,5龄前期的浓度比对照蚕低,5龄第3天几乎检测不到;全体照射与造血器官局部照射蚕之间的结果相似。照射蚕注射大肠杆菌后在体内诱导出现的应急蛋白量明显比对照蚕要少。由此认为家蚕幼虫造血器官与血淋巴中的蛋白质成分有关,造血器官的机能障碍、血球的数量减少可影响脂肪体中蛋白质的合成,从而使存在于血淋巴中的蛋白质成分发生变化。     

Effect of Temperature on Radiosensitivity of Newcastle Disease Virus

Newcastle disease virus was irradiated at temperatures ranging from 2.2 to 60 C. An interaction between the thermal and ionizing energy was observed in the temperature region of 49 to 60 C. At 2.2 C, the hemagglutinin was considerably more radioresistant than the infectivity property. It is believed that radiation inactivation of Newcastle disease virus infectivity at low temperatures was due to nucleic acid degradation and at higher temperatures was due to protein denaturation.     

EPR studies of 5-bromouracil crystal after irradiation with X rays in the bromine K-edge region

Radicals induced in a single crystal of 5-bromouracil (BrUra) by synchrotron soft X rays in the bromine K-edge region (13.461-13.482 keV) were investigated using the X-band EPR method. The crystal was irradiated at three peak energies of the absorption spectrum at room temperature or at 80 K. A hydrogen abstraction radical derived from N1 of the pyrimidine ring was commonly observed for all of the energies used, though with some variation in quantity. Similar characteristics were also observed in the EPR signal for the off-K-edge low-energy (13.42 keV) and (60)Co gamma rays used for comparison. When irradiated at 80 K, a much larger exposure (roughly 10 times) of soft X rays was needed to obtain the same signal intensity as that observed at room temperature. EPR signals were not detectable with gamma irradiation at liquid nitrogen temperature.     

辐照提高人工湿地美人蕉抗寒性及其越冬效果研究

植物是人工湿地的重要组成部分, 但大多数湿地植物在低温环境下不能生存, 微生物活性受到抑制, 导致人工湿地低温应用受到限制。研究将辐照育种的手段应用于湿地植物, 以提高其抗寒性。研究采用25、45、60和80 Gy的γ射线对美人蕉(Canna indica L.)种子进行辐照, 剂量率为6 Gy/min, 待幼苗长出后进行阶段性降温培养, 并评价各组抗寒性。结果表明: 在低温下, 辐照幼苗的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均高于对照组。对照组丙二醛(MDA)在低温下大量积累, 且含量显著高于辐照组。在低温条件下, 幼苗脯氨酸、可溶性糖、蛋白质含量高于对照组。辐照组抗寒性得分高于对照组, 45 Gy组得分最高。随后将最佳辐照处理的美人蕉种植于人工湿地, 对其冬季净化效果进行评估, 评价指标包括: 总磷(TP)、总氮(TN)、化学需氧量(COD)。结果发现, 45 Gy辐照组人工湿地的冬季TP去除率显著高于对照组(P<0.05)。由此可见, 辐照处理能够有效提高美人蕉的抗寒性, 并提高人工湿地冬季净化效果。     

~(16)O~(8+)离子辐照普通小麦引起幼苗可溶性蛋白质的不同变化

利用不同能量、不同剂量的1 6 O8+ 离子辐照春麦。处理方式为离子贯穿种子 (75MeV u)以及离子注入胚和胚乳等种子的不同部位。然后分析其幼苗的可溶性蛋白质 (solubleprotein) ,根据其分子量大小不同 ,将其划分为五个区段并计算各组分的相对含量 ,结果表明 :1 同对照相比 ,随着辐照剂量的增加 ,所有辐照材料 (包括贯穿和注入 )第二区段 (分子量较高区段 )蛋白质组分的相对含量下降 ;而第五区段 (分子量最小区段 )蛋白质组分的相对含量升高。 2 种子的贯穿处理 ,同时也引起第一区段 (分子量最高区段 )和第三区段蛋白质组分相对含量的下降以及第四区段的升高 ;其中第三、四区段的变化明显区别于注入效应。 3 注入胚和胚乳的区别在于后者第一区段蛋白质组分相对含量的较大下降和第五区段的较大提高。 4 小剂量辐照的材料 ,可溶性蛋白质组分的变化异常 ,可能与低剂量辐射兴奋效应有关。在讨论中提出了注入胚的即时效应与注入胚乳的后期效应。     

Influence of protein nutrition on dose-survival relationship following rat kidney irradiation

Immediately following unilateral nephrectomy the remaining kidney of juvenile male Sprague-Dawley rats was sham irradiated or irradiated to doses of 14-30 Gy. Following irradiation the animals were placed on isocaloric diets of either 20 or 4% protein. Median life spans for the animals on the low protein diet were significantly increased compared to the median life spans on the 20% protein diet. Serum urea nitrogen (SUN) levels were periodically measured in rats from each of the experimental groups. SUN levels in the irradiated rats fed the 20% protein diet increased significantly over unirradiated controls as a function of time. In contrast animals fed the 4% protein diet showed no significant changes in SUN levels irrespective of the size of radiation dose and time post irradiation. Renal protective factors calculated as the ratio of 80% survival times for animals fed the 20% protein diet compared to animals fed the 4% protein diet can be calculated to be 2.3 at 18 Gy and 2.8 at 22 Gy. Likewise, a SUN protective factor calculated as the ratio of percentage of nonirradiated control SUN values for the two diets (SUN 20% irradiated) (SUN 20% nonirradiated) (SUN 4% irradiated) (SUN 4% nonirradiated) is 2.4 for 18 Gy and 3.9 for 22 Gy.     

Photochromic pigments in akinetes and pigment characteristics of akinetes in comparison with vegetative cells of Anabaena variabilis

Since akinete germination is triggered by light and the action spectrum for this process has features in common with the spectra of the two photochromic pigments, phycochromes b and d, a search was made for the presence of these phycochromes in akinetes of the blue-green alga. Anabaena variabilis Kützing. Allophycocyanin-B was also looked for, since the action spectrum for akinete germination points to a possible participation of this pigment too. Isoelectric focusing was used for purification of the pigments. The different fractions were investigated for phycochromes b and d by measuring the absorbance difference spectra: for phycochrome b. 500 nm irradiated minus 570 nm irradiated, and for phycochrome d, 650 nm irradiated minus 610 nm irradiated. For determination of allophycocyanin-B. fourth derivative analysis of absorption spectra was made for some of the fractions from the isoelectric focusing column. Phycochrome b was also assayed for by measuring in vivo absorption difference spectra. The assays were positive for all three pigments. The complete photosynthetic pigment systems were also studied by in vivo fluorescence measurements on both akinetes and vegetative cells of Anabaena variabilis. Fluorescence emission and excitation spectra at selected emission wavelengths were measured at room temperature and liquid nitrogen temperature. The energy transfer from phycoerythrocyanin to phycocyanin is very efficient under all conditions, as is the energy transfer from phycocyanin to allophycocyanin at room temperature. At low temperature, however, phycocyanin is partly decoupled from allophycocyanin, particularly in the akinetes; the energy transfer from allophycocyanin to chlorophyll a is less efficient at low temperature in both types of cells, but especially in akinetes. Delayed light emission was measured for both types of cells and found to be very weak in akinetes compared to vegetative cells. From this study it would seem that akinetes lack an active photosystem II, although the 691 nm peak in the 570 nm excited low temperature fluorescence emission spectrum proves the presence of photosystem II chlorophyll, and also its energetic connection to the phycobilisomes.     

Decreased weight, DNA, RNA and protein content of the brain after neutron irradiation of the 18-day mouse embryo

Pregnant mice were irradiated with 0.5 Gy fission neutrons on the eighteenth day of their gestation. The average litter size at birth was unchanged but mortality increased 5-6 fold in the first 3 days. The irradiated mice were the same weight as control mice at birth but showed a progressively increasing weight deficiency up to at least 36 days as compared to controls. Brain weight was 37, 45 and 25 per cent less in 2-, 3- and 52-week old irradiated animals, respectively, and the ratio of brain weight to body weight was 25, 27 and 13 per cent less. The concentrations of DNA, RNA and protein (mg/g wet tissue) were the same in irradiated and control mice in both brain and liver at all three ages. Total DNA, RNA and protein contents of whole brain after irradiation were 56-75 per cent of the control levels. No definite decrease was observed in liver. Histological study at 6 hours after irradiation showed nuclear pyknosis in the central nervous system from definite to very severe according to the part examined. It is concluded that damage to the central nervous system of the 18-day mouse foetus after neutron irradiation is mainly due to killing and/or inhibition of the differentiation of neuroblasts.     

Reproduction of Japanese quail after microwave irradiation (2.45 GHz CW) during embryogeny

Japanese quail (Coturnix coturnix japonica) embryos were irradiated continuously in ovo with 2.45-GHz continuous wave radiation during the first 12 days of embryogenesis at an incident power of 5 mW/cm2 and a specific absorption rate of 4.03 mW/g. The internal temperature of irradiated and nonirradiated (sham) eggs was 37.5 +/- 0.3 degrees C, which is the optimum temperature for incubating quail eggs. At 35 days after hatching irradiated and sham-irradiated males were paired with irradiated or sham-irradiated females and daily records of reproductive performance were collected through 224 days of age. Progeny were hatched from each of the male-female pairs, and progeny reproductive performance was measured from 35 through 168 days of age. Hatchability was not affected by irradiation during embryogeny. Mortality after hatching, egg production, egg weight, fertility, hatchability of eggs produced, and reproductive performance of the progeny were not affected by irradiation during embryogeny. These observations indicate that irradiation of quail embryos with low-level microwave radiation does not affect the reproductive capacity of the hatchlings or of progeny produced from quail irradiated during incubation.     

Dose-response relationship between body temperature and birth defects in radiofrequency-irradiated rats

Five groups of pregnant Sprague-Dawley rats were irradiated for 10-40 min on gestation day 9 in a 27.12-MHz radiofrequency field at a magnetic field strength of 55 A/m and an electric field strength of 300 V/m. The specific absorption rate was 10.8 +/- 0.3 W/kg. Exposures were terminated after the rat's colonic temperature reached 41.0 degrees C, 41.5 degrees C, 42.0 degrees C, 42.5 degrees C, or 43.0 degrees C. A control group was sham irradiated at 0 A/m and 0 V/m on gestation day 9, whereas a second control group was untreated. The incidence of both birth defects and prenatal death was directly related to maternal body temperature once a temperature threshold was exceeded. The temperature threshold for both types of effects was approximately 41.5 degrees C. A few pregnant rats died after exposure to 43.0 degrees C, and higher temperatures were nearly always lethal.     

Immunological effects of amplitude-modulated radio frequency radiation: B lymphocyte capping

B lymphocytes collected from normal ICR Swiss mouse spleens were exposed in vitro in a Crawford cell to 147-MHz radiofrequency (RF) radiation, amplitude modulated by a 9-, 16-, or 60-Hz sine wave. The power densities ranged between 0.11 and 48 mW/cm2. The irradiated samples and the controls were maintained at 37 degrees C or 42 degrees C, with temperature variations less than 0.1 degrees C. Immediately after a 30-minute exposure, the distribution of antigen-antibody (Ag-Ab) complexes on the cell surface was evaluated at 37 degrees C by immunofluorescence. Under normal conditions (37 degrees C, no RF), Ag-Ab complexes are regrouped into a polar cap by an energy-dependent process. Our results demonstrate that the irradiated cells and the nonirradiated controls capped Ag-Ab complexes equally well after exposure at 37 degrees C. Capping was equally inhibited at 42 degrees C in both the controls and irradiated cells. No statistically significant differences in capping were observed between the RF-exposed and control samples at any of the modulation frequencies and power densities employed as long as both preparations were maintained at the same temperature.     

Relative biological effectiveness of the alpha-particle emitter (211)At for double-strand break induction in human fibroblasts

The purpose of this study was to quantify and to determine the distribution of DNA double-strand breaks (DSBs) in human cells irradiated in vitro and to evaluate the relative biological effectiveness (RBE) of the alpha-particle emitter (211)At for DSB induction. The influence of the irradiation temperature on the induction of DSBs was also investigated. Human fibroblasts were irradiated as intact cells with alpha particles from (211)At, (60)Co gamma rays and X rays. The numbers and distributions of DSBs were determined by pulsed-field gel electrophoresis with fragment analysis for separation of DNA fragments in sizes 10 kbp-5.7 Mbp. A non-random distribution was found for DSB induction after irradiation with alpha particles from (211)At, while irradiation with low-LET radiation led to more random distributions. The RBEs for DSB induction were 2.1 and 3.1 for (60)Co gamma rays and X rays as the reference radiation, respectively. In the experiments studying temperature effects, nuclear monolayers were irradiated with (211)At alpha particles or (60)Co gamma rays at 2 degrees C or 37 degrees C and intact cells were irradiated with (211)At alpha particles at the same temperatures. The dose-modifying factor (DMF(temp)) for irradiation of nuclear monolayers at 37 degrees C compared with 2 degrees C was 1.7 for (211)At alpha particles and 1.6 for (60)Co gamma rays. No temperature effect was observed for intact cells irradiated with (211)At. In conclusion, irradiation with alpha particles from (211)At induced two to three times more DSB than gamma rays and X rays.     

Change in protein synthesis and enzyme activity in avian endocrine glands during ontogenesis

The synthesis of cytoplasmic proteins and the activity of cAMP-dependent protein kinase were studied in the thyroid gland and thymus of intact and irradiated (0.05 Gr) embryos and chicks. The increase in labeled amino acid incorporation into proteins of irradiated chicken endocrine organs during postnatal development was more apparent in thyrocytes. Stimulation of protein kinase activity in the thyroid gland and thymus was observed at the same periods of time. It was supposed that minor doses of ionizing radiation stimulate synthetic processes as well as phosphorylation of proteins responsible for differentiation of avian endocrine organs.