Body X-irradiation Research Articles

Effect of two graded doses of x-irradiation on the rat adrenal gland and its protection by S-phenetyl formamidino 4(N-ethyl isothioamide) morpholine dihydrochloride

In the present study the amount of protection offered by the use of S-phenetyl formamidino 4(N-ethyl isothioamide) morpholine dihydrochloride to adrenal gland was investigated on albino rats against two graded doses of x-rays (1500R each). Total body x-irradiation brought about the hypertrophy and degranulation of adrenal cortical and medullary cells. The extent of hypertrophy and degranulation increased after the 2nd exposure to x-rays. Treatment with S-phenetyl formamidino 4(N-ethyl isothioamide) morpholine dihydrochloride prior to each dose of irradiation precluded the radiation changes caused in the adrenal cortex and medulla of the rats.

Anti-lipopolysaccharide toxin therapy for whole body X-irradiation overdose.

Death in humans from ionising radiation overexposure in the 3-8 Gy (300-800 rad) range is in part due to the toxaemia caused by the entry of gram-negative bacteria and/or their lipopolysaccharide toxin (LPS) into the blood circulation through the walls of partially denuded gut. Anti-LPS hyperimmune equine plasma was evaluated for its ability to lower irradiation-induced lethality. Mice were irradiated with 6.3 Gy (630 rad) and six days later received equine Anti-LPS hyperimmune plasma, control plasma or saline. Mortalities in the three groups were 58%, 92% and 79% (p less than 0.01) respectively. Thus Anti-LPS may prove useful as an adjunct to conventional therapy in treating radiation sickness.

Irradiation effects on the adrenal gland of rats undergoing inanition stress

The effect of total body x-irradiation was studied on rats under inanition stress. In response to irradiation an increase in the activity of cortex and medulla was noted in inanition stress administered rats rather than in the normally fed animals. Similarly, rising levels of urinary catecholamines and 5-hydroxytryptamine were observed in the starved animals after irradiation.

Reconstruction of lymph follicles in the mouse popliteal lymph node after X-irradiation damage.

In order to suppress possible "de novo" formation of follicles in the node after irradiation, animals to be irradiated received surgical operations to block the afferent lymphatics to the popliteal node on one side, the corresponding node on the other side left intact. 600 R whole body X-irradiation severely destroyed lymph follicles in the nodes of both sides, but the number of follicles in the node on either side recovered toward normal by 3 weeks after irradiation, regardless of whether they contained germinal centers or not. In animals exposed to 1,000 R whole body X-irradiation and infused with bone marrow cells, no lymph follicles were observed in the nodes on day 9. If such treated animals were given 1.0 X 10(8) lymph node cells on day 5, lymph follicles were reconstructed in the nodes of both sides on day 9. In animals irradiated with 1,000 R to the lower half of the body, the number of follicles in the node on either side remained comparable to that of the unirradiated control node. The results favor the interpretation that although X-irradiation destroys the lymphoid elements of the follicles, the stromata of follicles persist and preserve the ability to collect small lymphocytes, and that lymph follicles are reconstructed at the sites of the stromata persisting from damaged follicles.

The influence of timeshift on circadian rhythm of sensitivity to X-irradiation in mice.

For two groups of male C3H mice an eastbound transmeridional flight was simulated by inducing a time shift of the L:D schedule of 8 hr. The assumed flight brought about a maximal reduction of the daily light and dark span, respectively. A third group remained unshifted. At seven different times during the following day, subgroups of the time shifted mice as well as of the group with unchange schedule were exposed to whole body X-irradiation. Mortality and body temperature of each animal were registered for 30 days following exposure and were regarded as indicators of radiation response. Radioresistance was found to be highest during the second half of the daily light span, confirming earlier reports by other authors. Well defined effects of the time shift and a corresponding shift of the acrophase of radioresistance could be demonstrated. There was no significant difference between the two time shifted groups, but there was a consistent slight trend towards an advantage for the group whose L:D shift resulted in a maximally reduced dark span.

Haematopoietic role for Patch (Ph) revealed by newWmutant (Wct) in mice

SUMMARYA new mutant (Wct) has been identified at theWlocus of the mouse. The homozygote is poorly viable. Whereas the heterozygote (Wct/ +) is only mildly anaemic likeWυ/ +, the double heterozygoteWct+ / +Phis considerably more anaemic thanWυ+ / +Phand it andWsh+ / +Phhave significantly raised leucocyte counts.Wct+ / +Phis also unduly radiosensitive to whole body X-irradiation, 50% dying from haematopoietic failure at a dose of 4·59 ± 0·14 Gy, whereas the median forWct/ + was 6·49 ± 0·28 Gy. Serial blood counts of mice after low- or sub-lethal doses of X-rays revealed significantly more profound depression of counts of both red cells and leucocytes inWct+, and more notably inWct+ / +Ph, than in + / + orWsh/ + (haematologically normal) iso-dosed mice. We conclude that control of haematopoiesis by chromosome 5 is not confined to theWlocus but is shared by the linked genePh(and perhapsRw) and that expression of the change is not limited to the erythron but involves the pluripotent haematopoietic stem cell.

The action of azimexone on the cells of the hemopoietic system in mice, especially after damage with X-rays.

Mice were exposed to single doses of whole body X-irradiation (1 - 2 - 4 Gy) or were treated with sulphur mustard (15 mg/kg body weight i.p.). This treatment caused a reduction of the pluripotent stem cells in the bone marrow, of the total count of nucleated bone marrow cells in the femora and of the WBC in the peripheral blood. The size distribution of the bone marrow cells showed three separate peaks. From the histological examination of the bone marrow of X-irradiated mice it was deduced that the first peak represents erythrocytes, the second lymphocytes and the third peak the precursors of red and white blood cells. Multiple doses (25 - 50 - 100 mg/kg body weight) of azimexone, an immunomodulating substance, led after moderate doses of X-rays (2 Gy) or sulphur mustard to a more rapid recovery of the various parameters. In particular a stimulant action of azimexone on the pluripotent stem cells of mice not subjected to the injurious agents could also be demonstrated.

The Early Effect of Sublethal X-irradiation of Phagocytic Cells in Mouse Blood and the Influence of Cystamine as Measured by Chemiluminescence

The metabolic burst accompanying phagocytosis of granulocytes (PMN) leads to the generation of activated oxygen species such as O-2, H2O2, 1O2 and OH; which give rise to chemiluminescence (CL) in the presence of luminol. Reliable CL-measurements of stimulated PMN can be carried out in freshly drawn mouse blood, when photon counts are related to the number of PMN. Effects of low dose total body X-irradiation were studied using C57B1/6 mice. It was found that 24 and 48 hours after irradiation (0.24-0.95 Gy) CL of whole blood was slightly decreased. If however CL-counts were related to the number of PMN, an enhanced CL per single granulocyte was recorded. The administration of cystamine leads to an immune stimulating effect of unirradiated animals. In animals, who received 0.95 Gy a distinct radioprotective effect of cystamine can be observed.

Erythropoietin formation during hypoxia in mice with impaired responsiveness to erythropoietin induced by irradiation or 5-fluorouracil injection.

Plasma erythropoietin levels during continuous exposure to hypobaric hypoxia in mice with marrow aplasia induced by whole body X-irradiation of 5-fluorouracil injection were higher than in control mice similarly exposed. There findings give support to the hypothesis that a relationship exists between erythropoietin production rate and erythroid responsiveness to the hormone.

Functional modifications of macrophage activity after sublethal whole-body irradiation.

The effect of whole body x-irradiation on macrophage activity was evaluated by in vitro C3b-receptor-mediated ingestion and in vivo carbon clearance. Peritoneal macrophages from B/sub 6/D/sub 2/F/sub 1/ mice were tested from 20 min to 24 hr postirradiation and at doses up to 1000 rad x-irradiation. When compared to peritoneal macrophages from normal unstimulated mice, whole-body irradiation produced an increase in macrophage activity in both C3b-mediated ingestion and carbon clearance. The enhancement of macrophage activity was optimal at an exposure of 200 rad and waned as the exposure dose increased. Enhanced activity appeared as early as 20 min postirradiation and peaked by 3 hr.

Retrovirus particles in spontaneously occurring and radiation-induced tumors in ddY mice.

Among spontaneously occurring tumors, pulmonary tumor, malignant lymphoma, mammary tumor, and ovarian tumor were the major ones in 232 ddY female mice. The former three tumors appeared significantly earlier and the latter one increased in incidence in 229 mice given 600 R whole or 800 R partial body (tunk) x-irradiation at the age of 10 weeks. Five tumors were examined electronmicroscopically from each tumor type of both the spontaneously occurring and radiation-induced tumors. C type virus particles were present only in the malignant lymphoma and B particles in the mammary, ovarian and pulmonary tumors, which are all of epithelial character. Thus, as far as we were concerned in this mouse strain, B particles were produced preferentially in epithelial cells and C particles in non-epithelial cells. The retrovirus particles were found in the same frequency, namely, 10 of 20 tumors examined in both the tumor groups. From our results, the intervention of virus in radiation carcinogenesis still remains in question.

Investigations on the influence of whole body X-irradiation on the activity of rat lens aldose reductase (E.C. 1.1.1.21).

The influence of whole body X-irradiation on the activity of the enzyme aldose reductase (E.C. 1.1.1.21) in the lenses of young rats was investigated. Immediately after irradiation there was no difference between the activity of the enzyme in irradiated and nonirradiated animals. Within 10 days after irradiation, significant differences developed. There was a constant increase of the activity in the control lenses, while the activity in the irradiated lenses remained nearly constant. There was no remarkable difference between the two groups 10 days after irradiation with respect to the kinetics of the enzyme. This meant that irradiation does not affect the enzyme present at the moment of application, but the influence of 440 rad X-irradiation effects an obvious decrease in the synthesis of the enzyme.

Late effects of whole or partial body x-irradiation on mice: life shortening.

Life table data were obtained on 2400 ddY/SLC female mice exposed at 10 weeks of age to a single dose of X-rays. Doses given to the whole body were 95, 190, 380 and 570 rad, and to head, trunk or lower body, 190, 380, 570 and 760 rad. The mean survival time of unirradiated controls was 73 weeks. The mean survival times after whole body exposure decreased linearly with increasing dose. The estimate of life shortening per 100 rad was 7.2 per cent for whole body exposure. The life shortening after head or lower body exposure to a dose of 190 rad was 6.2 and 7.1 weeks respectively, and almost no further life shortening up to 760 rad. After trunk irradiation with 190 rad, life shortening was 10.4 weeks and beyond that dose life shortening was 1.1 per cent per 100 rad.

Aggregation patterns of fetal rat brain cells following exposure to X-irradiation.

In our search for a simplified in vitro test system to assess the teratogenic effects of physical factors, we studied the effects of total maternal body X-irradiation on aggregation patterns of enzymatically isolated fetal rat brain cells and on ultrastructural aggregate changes. The fetal brain cells were derived from day 14 gestation fetuses of pregnant Sprague-Dawley (CD strain) rats exposed to X-irradiation (25 - 200 R) one hour prior to sacrifice. Notable changes in the cell aggregates following X-irradiation included a reduction in cell aggregate size and an increase in number. The frequency of cell aggregates was higher in the treated than in the control group, and the mean diameter of cell aggregates was inversely related to increasing X-irradiation doses. Transmission electron microscopy revealed in isolated cells features of degenerative process which were similar to those found in intact fetal brain lesions caused by maternal X-irradiation. Furthermore, scanning electron microscopy revealed that inhibition of cell aggregation following X-irradiation could probably be attributed to inhibition of membrane filopodia development and a consequent failure fo cell aggregates to fuse into a greater cell aggregate mass. These results suggest that the membrane factors which influence cell aggregation may be a useful parameter to assess early effects of X-irradiation-induced brain deformity. Presently, the cell aggregation culture system is being further evaluated as a short term test system for environmental teratogens.

Normal serum reduces whole body X-irradiation enhanced growth of a mammary carcinoma in rats

Whole body X-irradiation (WBI) of experimental animals prior to transplantation of strongly immunogenic tumors frequently results in acceleration of tumor growth because of host immunosuppression. The growth of primary tumor ( p < 0.01) and metastatic tumor ( p = 0.006) can be enhanced by WBI (500–600 rad) of W/FU rats one day prior to inoculation with syngeneic ME/H mammary carcinoma cells that previously had been shown to be weakly- or non-immunogenic by usual excision-challenge methods. Spleen cells removed 1 day post WBI contained an increased proportion of neutrophilic granulocytes, increased capacity for in vitro cell mediated cytotoxicity, and caused enhanced growth of primary and metastatic tumor in the Winn assay. 34 Normal serum injections post WBI abrogated irradiation-enhanced tumor growth and irradiation-enhanced neutrophilic granulocytes. We suggest that both enhanced tumor growth in vivo and enhanced cell-mediated cytotoxicity in vitro are mediated by increased levels of neutrophilic granulocytes, presumably because of radiation destruction of a granulopoeisis inhibiting factor present in serum, which is restorable by normal serum.

ROLE OF MOBILE PASSENGER LYMPHOCYTES IN THE REJECTION OF RENAL AND CARDIAC ALLOGRAFTS IN THE RAT A PASSENGER LYMPHOCYTE-MEDIATED GRAFT-VERSUS-HOST REACTION AMPLIFIES THE HOST RESPONSE

It is demonstrated that passenger lymphocytes migrate out of rat renal allografts into host spleens in a radioresistant fashion. These mobile passenger lymphocytes within BN kidney and heart transplants are immunocompetent, since they elicit a graft-versus-host (GVH) reaction in the spleens of (LEW x BN)F2 hybrid hosts. The greater GVH reaction in (LEW x BN)F1 recipients of BN kidneys reflects the greater number of mobile passenger lymphocytes in the kidney when compared to the heart. The mobile passenger lymphocytes within BN renal allografts also cause a proliferative response in the spleens of LEW hosts as well as an accelerated rejection of BN renal allografts when compared to BN cardiac allografts, for the differences between BN kidney and heart, both in terms of splenomegaly elicited in LEW as well as tempo of rejection, are abolished by total body X-irradiation of the donor with 900 rad. Results indicate that a mobile passenger lymphocyte mediated GVH reaction in the central lymphoid organs of the host augments the host response to allogeneic kidneys and contributes materially to first-set renal allograft rejection; this GVH reaction on the other hand is not conspicuously present in LEW recipients of BN cardiac allografts and has therefore little effect on first-set cardiac allograft rejection.

Spleen cells of whole body X-irradiated W/Fu rats enhance tumor growth in vivo and non-specific cytotoxicity in vitro

This study was designed to investigate the influence of spleen cells of normal Wistar/Furth (W/Fu) rats obtained after whole body X-irradiation (WBI) upon mammary carcinoma growth in vivo, and cell mediated cytotoxicity against several target cells in vitro. The ME/H mammary carcinoma employed here originally arose spontaneously in a W/Fu rat, metastasizing to the retroperitoneal lymph node and lungs. It was found that surviving non-adherent spleen cells taken two days after 500R WBI cause enhanced tumor growth and metastases development in a Winn assay compared with nonadherent spleen cells from unirradiated controls. These cells were also enriched in granulocytes compared with controls. While the level of nonspecific cell mediated cytotoxicity was variable, it increased significantly following WBI of the spleen cell donor. Our results indicate that there are apparently two opposing effects shown by non-adherent spleen cells surviving WBI of normal W/Fu rats: enhancement of in vivo tumor growth, and enhancement of in vitro cell mediated cytotoxicity. A possible mechanism to explain these contrasting results is suggested.

Collection, cryopreservation and transplantation of blood stem cells in the treatment of hemopoietic failure.

It is the purpose of this paper to review the experimental studies which have been carried out in our group to explore the possibilities and limitations to treat hemopoietic failure induced by whole body x-irradiation by means of blood derived hemopoietic stem cells under autologous and allogeneic conditions. Some of the results have been published or are accepted for publication [1, 2, 5–17, 21]. The findings can be summarized in 5 statements supported by sufficient experimental evidence.KeywordsBlood Stem CellMononuclear LeukocyteHemopoietic Stem CellBone Marrow RecoveryBlood Derive Stem CellThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Short time observations of morphological changes and cholinesterase distribution in lymphatic organs of the mouse after corticosteroid and x-ray treatment

The changes in thymus, spleen and lymph nodes of the mouse after a single cortisone application or a single whole body x-irradiation were investigated morphologically and histochemically. During 24 h the alterations following the cortisone application are at all stages examined indistinguishable from those following the x-irradiation. The first signs of lymphocyte destruction can be observed already in the first two hours after treatment. Almost at the same time macrophages accumulate at the sites of cell death in the lymphatic organs studied. The eosinophils display a different behaviour. While they accompany the macrophages in the thymus already at the first stages, they appear in the spleen and lymph nodes with a latency of 6 and 8 h, respectively. The highest amount of necrotic cells is found ten hours after both treatments. At the same time the accumulation of macrophages and eosinophils reaches its maximum. The cholinesterase in the lymphatic organs is largely the true cholinesterase. The enzyme-activity increases in the cortex of the thymus gradually 6 h after treatment, showing the highest deposit of reaction product at 10 h. In spleen and lymph nodes the cholinesterase shows only slight variations. The possible role of the cholinesterase-activity in these non-cholinergic tissues is discussed.

Histophysiology of cellular immunity reactions in B-cell deprived rabbits. An X-irradiation model for delineation of an 'isolated T-cell system'.

Three times sublethal total body X-irradiation with thymus shielding--at 2 weeks' intervals--delineated a temporarily B-cell deprived animal model, only reconstituted with recently thymus-derived cells. The thymusdependent areas of peripheral lymphoid tissue-repleted with T-cells--are described. The cellular immune capacity of these animals with an "isolated T-cell system" was analyzed by means of skin allografting. Histological and autoradiographic studies were performed in draining lymph nodes after a variety of antigenic stimuli: skin allografts, S. java vaccin, horse-gamma-globulin, horse spleen ferritin and a contact sensitizer (Oxazolone).