Impact of Pulsed Nd:YAG Laser Irradiation on the Growth and Mortality of the Biofilm Forming Marine Bacterium Pseudoalteromonas carrageenovora

The impact of pulsed laser irradiation on the marine biofilm forming bacterium Pseudoalteromonas carrageenovora was investigated in the laboratory by monitoring mortality and the post-irradiation growth pattern. The impact of laser irradiation on bacterial mortality increased with the duration of irradiation. Laser irradiation at 532 nm (0.1 J cm m 2 ) for 15 min resulted in a 53% cell mortality immediately after irradiation. However, the impact after a period of 5 h (delayed impact) was more severe. The growth pattern of irradiated samples showed a prolonged lag phase compared to the reference, due to a reduction in total viable counts (TVC) in the irradiated samples. Nucleic acid staining is suggested to be a promising technique for monitoring laser inflicted bacterial mortality. Thus, the results suggest that laser irradiation could be considered as an alternative technique to reduce the number of biofilm forming bacteria and thereby biofilm formation on hard surfaces.     

Short- and long-term effects of whole-body irradiation with fission neutrons or X rays on the thymus in CBA mice

Young adult (6 weeks old) female CBA mice were exposed to whole-body irradiation with either 2.5-Gy fast fission neutrons of 1 MeV mean energy or 6.0-Gy 300 kVp X rays at centerline dose rates of 0.1 and 0.3 Gy/min, respectively. The weight of spleen and animal and the weight, cellularity, and histological structure of the thymus were studied at different times after irradiation. Thymic recovery after whole-body irradiation showed a biphasic pattern with minima at 5 and 21 days after irradiation and peaks of regeneration at Days 14 and 42 after X irradiation or at Days 14 and 70 after neutron irradiation. After the second phase of recovery, a marked decrease in relative thymus weight and cellularity was observed, which lasted up to at least 250 days after irradiation. Splenic recovery showed a monophasic pattern with an overshoot on Day 21 after irradiation. After neutron irradiation a late decrease in relative spleen and animal weight was observed. The observed late effects on thymus and spleen weight and thymus cellularity are discussed in terms of a persistent defect in the bone marrow.     

UV-induced imbalance of the deoxyribonucleoside triphosphate pool in E. coli

The effect of UV irradiation on the intracellular DNA precursor pool in E. coli was investigated. UV irradiation of E. coli, followed by post-incubation for 1-1.5 h, altered the relative sizes of the deoxyribonucleoside triphosphate (dNTP) pool. The total amount of dNTPs increased: both dATP and dTTP increased several-fold, dCTP about twofold, while dGTP remained almost unchanged. In recA- and umuC- strains, which are defective in UV-induced mutagenesis, the pattern of nucleotide pool alterations was similar to that of wild-type strains.     

The composition and physicochemical properties of the blood lipoproteins in rats exposed to external gamma irradiation]

Content of lipids, character of chemiluminescence of blood plasma and certain classes of lipoproteins have been studied. Geometrical parameters, nature and quantity of charged groups of lipoprotein particles accessible for titration have been determined 1 and 30 days after a single external gamma irradiation of rats in a dose of 3 Gy. The used irradiation dose exerts an expressed hyperlipidemic effect retained for one month after irradiation. The disturbances in the spectrum of blood lipids and lipoproteins are of hyper-beta and hyper-prebeta lipoproteinemia character. Considerable disturbances of physicochemical properties of different classes of lipoproteins have been detected. They are exhibited in changes of the pattern of free-radical processes, state of the charge of surface ionogenic groups and geometrical parameters of lipoprotein particles. Changes registered by the methods of potentiometric titration and correlation spectroscopy are most expressed in lipoproteins of very low density and those of low density.     

The reparative regeneration of the endothelium of the mouse aorta during microsurgical interventions following local gamma irradiation (based on scanning electron microscopic data)

In experiments with rats, abdominal aorta was subjected to microsurgical anastomosis after local irradiation with doses of 40 and 50 Gy. Irrespective of the time interval between the operation and irradiation the iatrogenic defect was restored completely. With the operation performed 24 h after irradiation the platelet adhesion decreased, the proliferation was inhibited depending on radiation dose, and the pattern of the endotheliocyte migration changed. The above effects were absent with the operation performed one month after irradiation.     

Normal development of the skeleton in chick limb buds devoid of dorsal ectoderm

It has been suggested that the ectoderm on the dorsal and ventral faces of the limb bud plays a part in controlling the pattern of cartilage differentiation. To test this, the dorsal wing bud ectoderm in the chick embryo was destroyed by irradiation with ultraviolet light at stage 17-19, at the very beginning of limb bud development, but the apical ectodermal ridge was spared. The irradiated ectoderm disappeared within 24 hr (by stage 23-24) and did not regenerate thereafter; thus the dorsal surface of the limb bud was kept denuded throughout most of the period of skeletal pattern formation. By 6 or 7 days after the irradiation (stage 35), when the rudiments of all the adult skeletal elements are normally present in recognizable form, the irradiated wings could be placed into two categories, those that were approximately normal in shape and those that had curled dorsally. All of these limbs were reduced in size, to varying degrees, when compared to their controls and lacked dorsal soft tissues. The limbs that were normal in shape, however, even though sometimes denuded over practically the whole extent of their dorsal surface, almost always had a complete and normally proportioned cartilage pattern, suggesting that ectoderm (other than the apical ectodermal ridge) does not exert any direct control over the development of the limb cartilage pattern. However, many of those limbs that had curled as a result of the irradiation did have major pattern deformities, suggesting that the topology of cartilage differentiation does depend on the shape of the limb bud.     

Repopulation of the mouse thymus after sublethal fission neutron irradiation. II. Sequential changes in the thymic microenvironment

The stromal cells of the thymus of sham-irradiated and sublethal fission neutron-irradiated CBA/H mice were analyzed with immunohistology, using monoclonal antibodies directed to I-A and H-2K antigens as well as specific determinants for cortical and medullary stromal elements. In the control thymuses, I-A expression in the thymus shows a reticular staining pattern in the cortex and a confluent staining pattern in the medulla. In contrast, H-2K expression is mainly confluently located in the medulla. Whole body irradiation with 2.5 Gy fission neutrons reduces within 24 hr the cortex to a rim of vacuolized "nurse cell-like" epithelial cells, largely depleted of lymphoid cells. The localization of I-A antigens changes in the cortex and I-A determinants are no longer associated with or localized on epithelial reticular cells. Medullary stromal cells, however, are more or less unaffected. A high rate of phagocytosis is observed during the first 3 days after irradiation. About 5 days after irradiation, the thymus becomes highly vascularized and lymphoid cells repopulate the cortex. The repopulation of the thymic cortex coincides with the appearance of a bright H-2K expression in the cortex which is associated with both stromal cells as well as lymphoid blasts. During the regeneration of the thymus, the thymic stromal architecture is restored before the expression of cell surface-associated reticular MHC staining patterns. The observed sequential changes in the thymic microenvironment are related to the lymphoid repopulation of the thymus.     

Influence of a 1400-gauss magnetic field on the radiosensitivity and recovery of EMT6 cells in vitro.

EMT6 mouse mammary tumour cells in vitro were exposed to an almost uniform 1400-gauss magnetic field during or after irradiation with 120 kV X-rays. Exposure of unirradiated control cultures to this field for up to 48 hours did not alter the viability or growth of the cells. Exposure of exponentially-growing cultures to the magnetic field during irradiation did not alter the survival curve. Exposure of exponentially-growing culturesto the magnetic field between two doses of 500 rad of X-rays did not alter significantly the pattern or magnitude of the recovery from sub-lethal damage. Exposure of plateau phase cultures to the magnetic field after irradiation did not alter significantly the pattern or magnitude of recovery from potentially-lethal damage. The effects of a short exposure to an almost uniform magnetic field of this strength on the production and repair of radiation damage appear to be minimal in this system.     

Development of the Xenopus laevis hatching gland and its relationship to surface ectoderm patterning.

An antibody that recognizes tyrosine hydroxylase can be used as a marker for hatching gland cells in Xenopus embryos. Using this marker, we have shown that hatching gland cells are induced at the end of gastrulation and that presumptive hatching gland cells are localized to the anterior neural folds in Xenopus. The movements of neurulation bring the hatching gland cells together to form a characteristic Y pattern on the dorsoanterior surface of the head. The Y pattern delineates several zones of surface ectoderm which can be visualized by the presence or absence of ciliated cells. As development proceeds the hatching gland pattern is altered, demonstrating the active changes involved in forming the face. Lithium, UV irradiation and retinoic acid can be used to alter the hatching gland pattern in specific ways which help to understand the underlying mechanisms of ectodermal patterning.     

Theoretical analysis of mixed irradiation (3).

As the model we proposed last year was contradictory to experimental data, we revised again the models for mixed irradiation by Zaider and Rossi and by Suzuki, substituting a 'reciprocal-time' pattern of repair function for a first-order one in reduction and interaction factors of the models, although we used a second order repair function last year. The reduction factor, which reduces the contribution of the square of a dose to cell killing in the models, and the interaction factor, which also reduces the contribution of the interaction of two or more doses of different types of radiation, were formulated by using the 'reciprocal-time' pattern of repair function. These newly modified models for mixed irradiation could express or predict cell survival more accurately than the older ones, especially when irradiation is prolonged at low dose rates. We present survival curves of cells calculated from the newly and the older models of assumptive simultaneous mixed irradiation with two or three types of radiation.     

Methylation changes of H19 gene in sperms of X-irradiated mouse and maintenance in offspring

The nature of imprinting is just differential methylation of imprinted genes. Unlike the non-imprinted genes, the methylation pattern of imprinted genes established during the period of gametogenesis remains unchangeable after fertilization and during embryo development. It implies that gametogenesis is the key stage for methylation pattern of imprinted genes. The imprinting interfered by exogenous factors during this stage could be inherited to offspring and cause genetic effect. Now many studies have proved that ionizing irradiation could disturb DNA methylation. Here we choose BALB/c mice as a research model and X-ray as interfering source to further clarify it. We discovered that the whole-body irradiation of X-ray to male BALB/c mice could influence the methylation pattern of H19 gene in sperms, which resulted in some cytosines of partial CpG islands in the imprinting control region could not transform to methylated cytosines. Furthermore, by copulating the interfered male mice with normal female, we analyzed the promoter methylation pattern of H19 in offspring fetal liver and compared the same to the pattern of male parent in sperms. We found that the majority of methylation changes in offspring liver were related to the ones in their parent sperms. Our data proved that the changes of the H19 gene methylation pattern interfered by X-ray irradiation could be transmitted and maintained in the first-generation offspring.     

Radiation-induced developmental abnormalities of the brain ventricles in the golden hamster

The dysontogenesis of the brain ventricle system of golden hamster fetuses was studied after X-ray irradiation of the mother at different stages of pregnancy. Groups of gravid hamsters were irradiated with a single dose (200 R) on either the 6th, 8th, 10th or 12th day of gestation. The fetuses were removed 1 day before birth. The most prominent finding in all stages of development was the dilatation of the brain ventricles. In addition to hydrocephalus, hematocephaly was also found in some cases. Furthermore, a widening of the central canal of the spinal cord with hemorrhagic content could sometimes be observed. Ependymal ectopies and subependymal rosettes occurred more frequently when irradiation was given in the late fetal period. The experimental findings demonstrate that after irradiation of the mother animal, malformation of the brain may be produced during the whole intrauterine period of development. The pattern and extent of malformation depend not only upon the dose of irradiation but also upon the stage of development at which irradiation was given.     

The reaction of rat blood plasma to gamma radiation studied by the use of laser photon-correlation spectroscopy]

Using the method of laser photon correlation spectroscopy it was shown that the typical pattern of distribution of blood plasma (BP) particles according to their sizes, characteristic of intact animals, was asymmetric with regard to a mean value. Gamma irradiation with sublethal doses was shown to change essentially the pattern of BP distribution. For instance, in BP of exposed animals, the number of particles of much smaller sizes increased as compared to that in intact animals. The smaller BP particle size was not the result of the direct effect of radiation on the blood. The in vitro irradiation of BP with various doses results usually in a considerable growth of particle sizes.     

Levels of liver glycogen during normal and radiation-induced metamorphosis in Rana cyanophlyctis

This study shows that significant changes in the pattern of glycogen deposition are brought about by irradiation with 2,000 R gamma-rays. The sharp fall during the 1st week can be attributed to its emergent mobilization. However, glycogen stored in the liver during the 2nd and 3rd week prior to the resorptive process substantiates the fact that tadpoles complete metamorphic transformations in spite of sublethal irradiation.     

Effect of irradiation on the size of isolated liver mitochondria

The role of external effects in the time course of changes in hepatocyte mitochondria size was studied. Suspensions of mitochondria isolated from the liver of inbred mice were investigated at various periods after gamma irradiation. Statistical analysis of changes in the size of isolated mitochondria has shown that irradiation leads to an increase in mitochondria size, reflected in the distribution pattern. The changes may be related to the functional activity, and distribution shifts may be regarded as manifestations of hepatocyte adaptation.     

Two types of proliferating cell nuclear antigen (PCNA) complex formation in quiescent normal and xeroderma pigmentosum group A fibroblasts following ultraviolet light (uv) irradiation.

We examined the relationship between the formation of proliferating cell nuclear antigen (PCNA) complex with DNA and nucleotide excision repair in human fibroblasts following ultraviolet light (uv) irradiation. PCNA complex formation was detected by the immunofluorescence method after methanol fixation and nucleotide excision repair activity was detected as the unscheduled DNA synthesis (UDS) by autoradiography labeled with [3H]thymidine. Quiescent normal cells showed a strong punctuated pattern of PCNA staining 5 min to 3 h and UDS 3 h after 10 J/m2 of uv irradiation, but they no longer showed PCNA staining and UDS 24 h after irradiation. In contrast, xeroderma pigmentosum group A (XP-A) cells, which lack UDS activity, did not show PCNA staining up to 30 min after irradiation; however, unexpectedly, they were stained 3 h and even 24 h after irradiation with their staining pattern being different from that in normal cells. Namely, the fluorescence spots in XP-A cells were larger in size and much smaller in number than those in normal cells. When XP-A cells were fused with normal cells with polyethylene glycol treatment, nuclei of XP-A cells showed a PCNA staining pattern similar to that of normal cells at 30 min, which was no longer detected 24 h after irradiation. These results suggest that there exist two types of PCNA complex formation, nucleotide excision repair-related and -unrelated, in human fibroblasts following uv irradiation.     

Expression of c-Fos gene in the rat hypothalamus upon electric painful stimulation and EHF irradiation of the skin

Studies of the activation pattern of hypothalamic cells upon electric painful stimulation have revealed an increase in the counts of c-Fos-positive cells (72.02-98.95%) predominantly of a size ranging from 10 to 50 microm2. EHF irradiation of the skin activates hypothalamic cells of the size ranging from 10 to 70 microm2 in the paraventricular, dorsomedial nuclei, and in perifonical zone of the lateral hypothalamic area. The combined action of EHF skin irradiation and electric painful stimulation leads to a decrease in the counts of cells activated in result of the painful stimulation in the anterior hypothalamic area, ventromedial nucleus, and the basal part of the lateral area. Changes in size distributions of activated cells are found in the ventromedial nucleus and the basal part of the lateral area, occur mainly through decreased counts of smaller cells (10-30 microm2) and increased counts of lager cells. The modulatory effect of EHF irradiation of the skin was found to be expressed as a reduction in hypothalamic cell activation caused by electric painful stimulation.     

Establishment of the dorsal/ventral polarity of the amphibian embryo: Use of ultraviolet irradiation and egg rotation as probes

Ultraviolet irradiation and egg rotations were employed as probes for the study of the establishment of the dorsal/ventral polarity of the amphibian embryo. Ultraviolet irradiation was discovered to alter the natural position of the doral lip and to modify the pigmentation pattern of the early embryo. Rotation of the uncleaved egg was found to succeed in relocating the dorsal lip to a new site in the embryo. Also, rotation of the egg was capable of preventing the characteristic defects associated with irradiation of the uncleaved egg. A combination of these probes was employed, and the results were interpreted in terms of models for the role of the egg surface and the internal cytoplasm in the establishment of the dorsal/ventral polarity of the egg.     

The mechanism of differential sister chromatid fluorescence as studied with the GC-specific DNA-ligand mithramycin

Chromosomes of human blood cells exposed to BUdR for two cell cycles showed an R-band pattern of fluorescence without lateral differentiation after staining with the GC-specific DNA-fluorochrome mithramycin. Differential sister chromatid fluorescence could be induced by a mild near-ultraviolet irradiation pretreatment which was without effect in Feulgen staining. Thus, besides the primary alteration of DNA structure caused by incorporation of BUdR, secondary structural alterations, probably mediated via chromosomal proteins, are required in order to obtain differential mithramycin-fluorescence of sister chromatids. The differential staining pattern was similar to that achieved with the AT-specific DNA-fluorochrome DAPI. Therefore, it may be concluded that the base specificity of fluorochromes does not play any part in the production of differential fluorescence of sister chromatids by this method.     

Cell cycle traverse in NHIK-3025 carcinoma of the uterine cervix after low-dose-rate irradiation

Theoretically, fractionation schemes could be tailored to the individual pattern of radiation-induced synchronization of cells in the radiosensitive G2 phase, leading to more effective radiotherapy. Using a human cervical carcinoma xenografted to nude mice, the effects of low-dose-rate irradiation on the cell cycle distribution were studied. Flow cytometric analysis demonstrated that cells accumulated in G2 + M phase 35 h after a total dose of 10 Gy of 137Cs irradiation. This accumulation time corresponded closely to the cell cycle time (Tc) (31 h) of this tumor, as determined by autoradiography. Further experiments are planned to determine the potential of fractionation schemes adjusted to the Tc-related accumulation in G2 in improving the effectiveness of radiotherapy.