固定化生长细胞生物催化的若干动态性质

通过啤酒发酵实验研究了以海藻酸盐为载体的固定化生长细胞生物催化作用的若干动态特性．在分批出料的填充柱循环反应器中，产物抑制型的啤酒发酵、其Ｓ～ｔ，Ｐ～ｔ曲线与批式发酵罐的Ｘｔ／Ｘｓｔ～Ｋｔ理论曲线相似，Ａ（λ＝５２０ｎｍ）的变化可作为简便的检测方法．发酵周期的缩短呈现τｎ＝ｎ－１（τ１－Ｃ）关系，式中ｎ＝２～４．这对生产上前期的成功应用是有力的支持．但在ｎ≥５时出现载体的崩解和起泡现象，因相催化剂不能继续使用．     

INTERACTION OF RESTORATION PROCESSES IN UV IRRADIATED ESCHERICHIA COLZ CELLS

Abstract— An attempt was performed to estimate survival and course of DNA synthesis in Escherichia coli B/r hcr' and hcr^- cells in relation to the amount of unexcised dimers.
In exponential growing hcr⁺ cells irradiated with 30 Jm^-2, dimers were almost completely excised and survival of cells was equal to about 3%. In the hcr⁺ cells prestarved for amino acid and thymine and irradiated by the same fluence, survival of cells was almost equal while two thirds of dimers remained unexcised and could be detected in the hybrid DNA consisting of parental and daughter chains. In exponentially growing hcr^+- cells irradiated with 20Jm^-, the same amount of dimers was produced which remained unexcised in the prestarved hcr⁺ cells. However, their survival was equal to about 0.02%.
Despite the great differences in dimer contents, about one third of DNA was replicated after UV in both exponentially growing and prestarved hcr+ cells producing well defined HL-hybrid peak, and the newly synthesized DNA was normal-sized. In hcr⁺ cells which contained approximately the same amount of dimers as in hcr⁺ prestarved cells, the amount of replicated DNA was too low to form a detectable density labelled hybrid peak, and the newly synthesized DNA was in short pieces.
Thus, when hcr⁺ and hcr^+- cells contain the same number of residual dimers, they have different levels of tolerance to these dimers.     

KINETICS OF PHOTOREACTIVATION AND LIQUID-HOLDING RECOVERY IN YEAST CELLS

Abstract— A method is presented for analyzing data on the kinetics of photoreactivation and liquid-holding recovery in microorganisms. The method extracts, from measurements on survival, the number of repairable lethal hits per cell remaining after a period of photoreactivation or liquid-holding recovery. A semilogarithmic plot of this quantity as a function of the duration of the survival-enhancing treatment reveals the nature of the kinetics of inactivation of the lethal hits.
The method has been applied to photoreactivation and liquid-holding recovery in yeast cells with wild-type radiation resistance. In the case of photoreactivation of ultraviolet-damaged cells, the number of lethal hits per cell is found to decrease exponentially with the length of exposure to a continuous source of photoreactivating light. Liquid-holding recovery in cells exposed to ultraviolet light or X-rays also results in exponential decrease of lethal hits with increasing time.
The method is compared with the fluence-decrement method and Dulbecco's treatment of photoreactivation data.     

METABOLISM OF tRNAs IN GROWING CELLS OF Escherichia coli ILLUMINATED WITH NEAR-ULTRAVIOLET LIGHT

Abstract— The tRNA metabolism which accompanies illumination of growing E. coli cells has been examined in conditions that led to growth delay. (i) The in vivo formation of the 8–13 link was followed by a fluorimetric procedure and revealed pseudo-first order kinetics very close to those obtained in vim under the same illumination conditions. The yield of 8–13 link appears to be quantitative (± 10%). Comparison of these kinetics with the radiochromatographic data of Blanchetot et al . (1984) suggests the transient formation during illumination of a new RNase-T,-resistant dinucleotide in tRNA distinct from the 8–13 link. (ii) Evidence is provided that under illumination some tRNA molecules lack one or more bases in a specific position in the sequence, thus yielding discrete fragments after aniline treatment. (iii) During the growth lag, uracil incorporation into nucleic acids occurs at an apparent rate between 4–8% of that normally observed during exponential growth. Evidence is provided however that the pyrimidine ribonucleoside triphosphate pools are strongly perturbed after illumination. Comparison of exogenous [³H]uracil incorporation into two strains proficient or deficient in uracil biosynthesis suggests a derepression of the endogenous path after light treatment. In addition, the UTP-to-CTP conversion is inhibited. In spite of preferential incorporation of exogenously labelled uracil in tRNA after illumination, a possible pyrimidine base turnover cannot be proved. These data are compatible with tRNA repair (Blanchetot et al ., 1984) involving a few tRNA species.     

MAINTENANCE OF DNA REPAIR CAPACITY IN DIFFERENTIATING RAT MUSCLE CELLS IN VITRO

Abstract— Unscheduled DNA synthesis has been measured at several times during the differentiation of cultured rat skeletal muscle cells in response to exposures to 254nm UV light. There is no change in the amount of repair DNA synthesis as the cells fuse and differentiate from postmitotic prefusion myoblasts to multinucleated contracting myotubes.     

PHOTOREACTIVATION OF ULTRAVIOLET IRRADIATED NON-DIVIDING POPULATIONS OF ICR 2A FROG CELLS

Abstract— Ultraviolet (UV) irradiation of non-dividing populations of ICR 2A frog cells led to their detachment from the surface of the culture dish and eventual lysis. Exposure of the cells to photoreactivating light after UV irradiation prevented cell killing and was accompanied by a loss of endonuclease sensitive sites from DNA. This photoreversal did not take place when the cells were exposed at 4°C to photoreactivating light indicating that the reversal was the result of photoenzymatic repair. As the action of photoreactivating enzyme is specific for the repair of pyrimidine dimers in DNA, these results suggest that pyrimidine dimers in DNA are the critical lesions leading to the death of non-dividing populations of UV irradiated cells.     

PHOTOMIMETIC EFFECT OF SEROTONIN ON YEAST CELLS IRRADIATED BY FAR-UV RADIATION

Abstract— The effect of serotonin on the survival of far-UV irradiated cells of the yeast Candida guillier-mondii was studied. Serotonin was found to have a photomimetic property. Preincubation of cells with serotonin results in protection against far-UV inactivation, whereas the post-radiation treatment with serotonin causes a potentiation of far-UV lethality. Both effects are similar to those produced by near-UV (334 nm) radiation. The observations provide support to the idea advanced by us previously that photosynthesized serotonin is the underlying cause of the two effects of near-UV radiation, photo-protection and potentiation of far-UV lethality. Experiments with an excision-deficient strain of the yeast Saccharomyces cerevisiae suggest that the effect of serotonin is by its binding to DNA.     

OVERPRODUCTION OF SSB PROTEIN ENHANCES THE CAPACITY FOR PHOTOREPAIR IN Escherichia coli recA CELLS

We studied photoreactivation in cells carrying the multicopy ssb+ plasmid. These cells overproduce single-stranded DNA-binding protein (SSB). Overproduction of SSB enhances the capacity for photoreactivation in recA bacteria but not in the recA+ background. It is suggested that, in recA cells, SSB binds to the dimer region of DNA and that this binding stimulates the process of photoreactivation. In recA+ cells, the same stimulation might be achieved by RecA protein.     

METABOLISM OF NUCLEIC ACIDS IN U.V. IRRADIATED L-STRAIN CELLS TREATED WITH NATIVE DEOXYRIBONUCLEIC ACID

Abstract— In our earlier papers we described the recovery of U.V. irradiated L-strain cells by means of nucleic acids. In an attempt to get better insight in the mechanism(s) leading to this recovery we studied the effect of native DNA on the metabolism of nucleic acids of recipient cells. In these experiments we have found that the label isologous DNA-C¹⁴ is incorporated in DNA of normal and irradiated recipient cells. The rate of DNA-C¹⁴ label incorporation in the DNA of irradiated recipient cells is slower than in the unirradiated controls. The presence of thymidine in the incubation media reduces the incorporation rate of DNA-C¹⁴. When the irradiated cells have been growing in a medium containing isologous DNA in presence of thymidine or heterologous DNA the recovery effect was not observed.     

SITES SENSITIVE TO S1 NUCLEASE and DISCONTINUITIES IN DNA NASCENT STRANDS OF ULTRAVIOLET IRRADIATED MOUSE CELLS

Abstract Mouse 3T3 cells irradiated with ultraviolet light synthesize DNA containing sites sensitive to the single-strand specific SI nuclease. The appearance of these sites correlates well with the presence of discontinuities in nascent strands, detected by the methodology of sedimentation in alkaline sucrose gradient. Thus, both the sites sensitive to SI nuclease and the discontinuities in nascent strands (i) are stabilized by caffeine; (ii) are no longer formed late after irradiation and (iii) disappear faster when a certain UV fluence is split into two fluences whose sum equals the single fluence. Moreover, the recovery in synthesizing DNA without SI sensitive sites is not dependent on excision repair of pyrimidine dimers or on continuous DNA synthesis. These SI sensitive sites are exclusive of replicative structures of irradiated cells and should correspond to stretches of single-strand DNA (gaps) formed during replication.     

RECOVERY FROM INHIBITION BY UV-IRRADIATION OF ORNITHINE DECARBOXYLASE INDUCTION IN HUMAN CELLS: IMPLICATION OF EXCISION REPAIR

Abstract— Exposure of stationary-phase human breast carcinoma(T–47D) cells to far-UV light (254 nm) inhibited the appearance of induced ornithine decarboxylase (ODC) activity. The fluence response curve had a shoulder (D₄= 2 J m_-2) followed by an exponential decline (D₀= 4.2 Jm_-2). The cells could recover from this inhibition when the stimulus of induction of ODC was delayed for20–24 h after irradiation. Hydroxyurea (HU) when present at 3 mM during the recovery period eliminated completely the ability of the cells to recover. This effect of HU on ODC induction was partially reversed by 50 nM of the four deoxyribonucleosides required for DNA synthesis. Neither HU nor the deoxyribonucleosides by themselves affected ODC induction in unirradiated cells. Since HU inhibited the recovery from potentially lethal UV damage and is a known inhibitor of excision repair, we interpret the above results to mean that recovery from UV-induced inhibition of ODC induction depends on excision-repair of DNA damage. This interpretation is strongly supported by the finding that specific photolysis of 5-bromodeoxyuridine, incorporated into DNA during the recovery period, inhibited recovery of ODC induction from inhibition by UV light.     

RECOVERY OF SUBCHROMOSOMAL DNA SYNTHESIS IN SYNCHRONOUS V-79 CHINESE HAMSTER CELLS AFTER ULTRAVIOLET LIGHT EXPOSURE

Abstract— Previous work obtained from Chinese hamster V-79 cells indicated that, immediately following exposure, UV-induced lesions acted as blocks to elongation of nascent strands, but gradually lost that ability over a 10 h period after exposure to 10 J/m². The work reported herein attempted to examine possible cell cycle mediated alterations in the recovery of DNA synthesis. Kinetic incorporation of radiolabeled thymidine studies indicated that there may have been a more rapid recovery of DNA synthesis in cells irradiated in G₁ or G₂ vs cells irradiated in S phase. DNA fiber autoradiograms prepared from synchronous cells indicated that after irradiation in any phase of the cell cycle, the length of newly synthesized DNA was equal to control lengths 1 h after exposure to 5.0 J/m² (or 1 h after entering S phase for cells irradiated in G₁ or G₂). This observed recovery was not solely due to an excision process. No cell cycle mediated difference in the number of dimers induced or removed as a function of cell cycle position was observed. These results appear to be consistent with a continuum of effects, with initiation effects dominating the response at low fluences, gapped synthesis at intermediate fluences and elongation inhibition at high fluences. The fluences at which each event dominates may be cell-line specific.     

COMPARATIVE POTENCY OF DIFFERENT UV SOURCES IN REDUCING THE DENSITY AND ANTIGEN-PRESENTING CAPACITY OF LANGERHANS CELLS IN C3H MICE

Abstract Although broadband UV-B irradiation has been shown to induce selective immunosuppression in a variety of experimental systems, the wavelength dependence of the immunomodulation and the initial events in the skin remain unclear. In the present study three UV lamps were used at suberythermal doses on C3H mice: a conventional broadband UV-B source (270–350 nm), a narrowband UV-B source (311–312 nm) and a UV-A source (320400 nm). Their effects on the photoisomerization of the naturally occumng trans- isomer of urocanic acid (UCA) to cis- UCA, on the density of Langerhans cells and on the ability of epidermal cells to stimulate allogeneic lymphocytes in the mixed skin lymphocyte reaction (MSLR) were ascertained. Broadband UV-B irradiation was more efficient than narrowband UV-B at reducing the density and function of Langerhans cells, while UV-A irradiation was least effective. These changes were most pronounced immediately following irradiation, were dose dependent and were only detected in UV-exposed areas of skin. There was a close correlation between the UV-induced reduction in Langerhans cell density and the formation of cis -UCA in the epidermis. This correlation was not detected between the reduction in the MSLR response following UV irradiation in vivo and cis-UCA formation.     

DNA REPAIR IN ULTRAVIOLET LIGHT IRRADIATED HeLa CELLS AND ITS REVERSIBLE INHIBITION BY HYDROXYUREA

Abstract— –The repair of u.v. damaged DNA in HeLa cells can be detected using the alkaline sucrose gradient technique. As a result of pyrimidine dimer excision single strand breaks are produced in DNA of irradiated cells. Rejoining of these breaks occurs during an 8 hr post-irradiation incubation period and is prevented by hydroxyurea and acriflavine. The inhibition of repair by hydroxyurea can be reversed by a mixture of all 4 deoxyribonucleosides at a concentration that does not reverse the inhibition of total DNA synthesis.     

TWO-PHOTON INACTIVATION, PHOTOREACTIVATION AND PHOTOPROTECTION IN YEAST CELLS IRRADIATED BY 266 NM-LASER RADIATION

Abstract— The quadratic dependence of inactivation of yeast cells on laser fluence rate at 266 nm suggests the occurrence of two-photon photochemical reactions in DNA, producing photoproducts similar to those found with ionizing radiation. Observations of a significant diminution of photoreacti-vation and the disappearance of photoprotection at high laser fluence rates provide additional evidence for this.     

辐照肉制品中2-十二烷基环丁酮的测定

采用GB/T 21926-2008《辐照含脂食品中2-十二烷基环丁酮测定气相色谱/质谱法》对辐照肉制品中的2-十二烷基环丁酮进行测定,鉴别辐照肉制品。样品用索氏抽提,经冷冻离心和弗罗里硅土层析净化,用气相色谱质谱联用法测定。2-十二烷基环丁酮的浓度在0.01~0.5 mg/L与色谱峰面积呈良好的线性关系,r=0.999 6。对未经辐照的样品进行加标回收试验,加标回收率为83.1%~94.0%,测定结果的相对标准偏差小于6%(n=6)。     

DECAY KINETICS OF FREE RADICALS IN IRRADIATED PROTEINS

Abstract— A detailed analysis is presented of the decay kinetics of free radicals in irradiated biopolymers. Available data in the literature are critically evaluated and new data for irradiated keratin are presented. It is shown that a rigorous analysis of the data available in the literature leads to more detailed information on the decay mechanisms than the conclusions which have been previously given; in some cases the analysis leads to mechanisms contradicting those claimed in the literature.
Our results suggest that the free radicals in illuminated keratin consist of three distinct species: one of the species decays with a half life of approximately 20 hr, the second has a half life of about 4600 hr, while the half life of the third is so long that the free radical concentration is practically constant. A similar behaviour is obtained on assumption of second order decay.
The consistency of the kinetic constants obtained has been verified in kinetic experiments using samples prepared under very different experimental conditions.     

ESR STUDY OF FREE RADICALS IN IRRADIATED POLYAMIDE-1010

Polyamide-1010 samples were irradiated in vacuum at room temperature by Cobalt-60 γ-rays. The free radicals formed in irradiation were studied by means of electron spin resonance (ESR)techniques.The ESRspectra consisted of a quartet and a superimposed singlet which were attributed to radical -CO-NH-CH-CH_2 and -CH_2-C=O, respectively. The effects of temperature and crystaUinity on the radicals were discussed and the mechanism for the production and decay of the radicals was also proposed.     

SIMULTANEOUS GROWTH AND DECAY OF FREE RADICALS IN IRRADIATED PROTEINS

Abstract— The electron paramagnetic resonance signals due to free radicals created in illuminated keratin have been studied during illumination and also during the decay of the signal in the dark. A number of mechanisms suggested to account for the simultaneous growth and decay of radicals during illumination is considered but only one of these is capable of accounting for the experimental results.     

CONSECUTIVE FORMATION OF SENSORY PHOTOSYSTEMS IN GROWING Halobacterium halobium

Abstract— The sensory photosystems PS 370 and PS 565 of Halobacierium halobium are actively degraded in the early growth phase and later resynthesized. Neither degradation nor resynthesis is correlated to the rate of cell division. The reappearance of photosensory activity requires de novo synthesis of proteins which are most likely directly involved in the sensory pathway. PS 370 appears earlier than PS 565 and thus may be studied independently of PS 565, before the latter is synthesized, or by blocking the synthesis of PS 565 with puromycin after PS 370 has appeared. The action spectrum of PS 370 alone shows the same maximum as the spectrum obtained when PS 565 is present. Carotenoids, which act as accessory pigments of PS 370, do not shift its activity peak. Also the maximum of PS 565 is not influenced by PS 370. We conclude that the maxima of the action spectra of PS 370 and PS 56.5 truely reflect the absorption maxima of the sensory retinal pigmentsP–370 andP–565.