Effect of continuous irradiation with a very low dose of gamma rays on life span and the immune system in SJL mice prone to B-cell lymphoma.

Ionizing radiation has been shown to have dose- and dose-rate-dependent carcinogenic effects on the hematopoietic and lymphoreticular systems. We report here that continuous exposure to a low dose of gamma rays influences the course of spontaneous B-cell lymphoma in SJL mice. We studied the biological effects of 10 cGy year(-1) gamma rays on the life span of 560 4-week-old SJL/J female mice and on various parameters of the cell-mediated immune response. Life span was slightly prolonged. The mean survival was 397 days for controls and 417 days for irradiated mice that died with lymphoma (P = 0.34). In lymph nodes and spleen, lower percentages of CD4+ and CD8+ T cells were observed in irradiated mice before 32 weeks. Interestingly, the percentages of CD49+ NK cells were increased in the spleens of irradiated mice at 28 weeks (0.61 +/- 0.08% compared to 0.43 +/- 0.12% in controls, P = 0.01) and at 32 weeks (0.62 +/- 0.24% compared to 0.33 +/- 0.09%, P = 0.02), while NK cell activity remained unchanged in exposed mice. These results provide further support for the absence of harmful effects of a continuous very low dose of radiation on life span and incidence of lymphoma in SJL mice.

Life span, cancer and non-cancer diseases in mouse exposed to a continuous very low dose of gamma-irradiation.

PURPOSE: To analyse the life-span and pathologies of mice living under a continuous very low-dose gamma-irradiation. MATERIAL AND METHODS: We exposed 300 C57B1/6J female mice, 3 weeks old, to 10 cGy year(-1) gamma-rays while 300 control mice lived in the same room. Irradiation was delivered continuously by thorium nitrate. We kept all the animals until natural death and performed autopsy. RESULTS: No difference was observed in life-span (mean lifespan +/-SE: 805.2 +/- 9.62 days for controls and 815 +/- 9.57 days for irradiated mice), weight curves or food intake. At autopsy, cancer was present in 40.9% of controls and 37.9% of irradiated mice. They were mainly represented by lymphomas (23.7 and 24.9%) and histiocytic sarcomas (12.6 and 8.7%, respectively, for controls and irradiated mice). Vascular diseases occurred in 24.1% of controls and 23% of irradiated mice. Infections were present at autopsy in 14.1 and 12.3%, respectively, of controls and irradiated animals. No statistical difference was observed at the end of the experiment for cancer or non-cancer diseases between the two groups. CONCLUSION: Continuous 10 cGy year(-1) gamma-irradiation had no adverse effect on malignant or non-malignant diseases in this strain of mouse.

Influence of continuous, very low-dose gamma-irradiation on the mouse immune system.

PURPOSE: To investigate whether continuous, very low-dose gamma-irradiation (10 cGy/year) modifies immune parameters in mice. MATERIAL AND METHODS: C57BL/6 female mice, 4 weeks old, were irradiated for 24 months and compared with control mice living in the same room. B- and T-cell subsets were evaluated by flow cytometry before and after stimulation with lectins; subclasses of immunoglobulins were determined by ELISA 2, 4, 6, 8, 12, 18 and 24 months after the beginning of the irradiation. RESULTS: No difference was found in the percentage of CD4(+) and CD8(+) cells in the thymus and the spleen, or in the reactivity of T-cells to lectins. While the number of B-cells in the spleen remained unchanged, a significant decrease of IgG1, IgG2b and IgG2a was observed after respectively 12, 18 and 24 months of irradiation. CONCLUSION: The parameters of cellular immunity studied were not affected by this chronic low-dose of irradiation, but this dose rate is probably too low to induce the hormetic effect previously described. Further investigations are necessary to assess whether the decline of immunoglobulin secretion is indicative of a lower rate of infectious diseases or a defect in B-cell function.

Reduced expression of retinoic acid receptor beta protein (RAR beta) in human papillary thyroid carcinoma: immunohistochemical and western blot study.

AIMS: To investigate the role of retinoic acid receptor beta (RAR beta) in thyroid carcinogenesis, we have investigated its expression in human thyroid samples by combined immunohistochemistry and Western blotting. METHODS AND RESULTS: Fifty-eight paraffin-embedded thyroid samples (40 normal or benign tissues, 16 papillary and two follicular carcinomas) were analysed by immunohistochemistry using a specific monoclonal antibody. Western blotting was also carried out on 11 selected samples (seven normal or benign tissues, three papillary carcinomas and one follicular carcinoma) and two human ovarian carcinomas as controls. RAR beta immunostaining was nuclear and limited to the normal epithelial thyroid tissue. A dramatic decrease in RAR beta immunostaining was observed in all the papillary carcinomas and in one follicular carcinoma. The other follicular carcinoma exhibited strong RAR beta immunostaining. By immunoblotting, a 51 kDa signal corresponding to the RAR beta was observed in nuclear extracts from normal thyroids and for one follicular carcinoma. However, this signal was lacking in the papillary carcinomas. These results were in complete agreement with the observations obtained by immunohistochemistry on the same samples. CONCLUSION: We present here the first demonstration of RAR beta protein in normal human thyroid follicular cells. In addition, we found that its expression is decreased in papillary thyroid carcinoma.

Clinical characteristics and light and transmission electron microscopic sperm defects of infertile men with persistent unexplained asthenozoospermia.

OBJECTIVE: To determine, with the use of transmission electron microscopy (TEM), the proportion of patients with permanent unexplained asthenozoospermia (<30% motility) who have an abnormality of sperm axonemal and periaxonemal structures. DESIGN: Retrospective study. SETTING: A university-affiliated public hospital. PATIENT(S): Sixty-one infertile men whose semen was submitted to TEM analysis because of persistent unexplained asthenozoospermia. MAIN OUTCOME MEASURE(S): The results of quantitative TEM analysis of the tails of the spermatozoa. INTERVENTION(S): None. RESULT(S): Based on a comparison with the axonemal anomalies observed in nine fertile control patients, the infertile population was divided into three groups: group I, with no detectable axonemal defects (26.2%); group II, with axonemal anomalies in either the midpiece or the principal piece (29.5%); and group III, with axonemal anomalies in both the midpiece and the principal piece (44.3%). However, defects in the mitochondrial sheath, fibrous sheath, and sperm head (acrosomic and postacrosomic cap) were observed in at least 50%, 30%, and 50%, respectively, of the patients in each group. The proportion of dense fiber anomalies of the midpiece increased significantly from group I to group III. No differences were observed between the three groups in sperm characteristics, anamnesis information, or clinical data. CONCLUSION(S): In patients with persistent unexplained asthenozoospermia, the frequent association of periaxonemal anomalies with axonemal deficiencies strongly suggests that axonemal deficiencies are not the unique cause of decreased motility.

Effect of a continuous gamma irradiation at a very low dose on the life span of mice.

BACKGROUND: There is epidemiological evidence that suggests there are beneficial effects of ionizing radiation at low doses. Some experimental studies confirmed this hormetic effect with doses of about 1 cGy/day, but no data concerning very low dose rates are available. OBJECTIVE: The aim of this study was to determine the life span of mice exposed to very low doses of ionizing radiation. METHODS: Six hundred female C57BL/6 mice, 1 month old, were exposed to chronic gamma irradiation at very low dose rates of 7 or 14 cGy/year. These doses are about 25 or 50 times higher than background, but much lower than the doses of about 1 cGy/day used in previous experiments. Three hundred mice living in the same room were used as controls. RESULTS: The life span, after the beginning of the experiment, determined by the survival time of 50% of each population, is increased in irradiated mice: 549 days in controls, 673 days in both irradiated groups. The differences are significant between the control and the irradiation mice. Differences between mice irradiated with 7 or 14 cGy are not significant. CONCLUSIONS: These results confirm the possibility of a nonharmful effect (hormesis) of ionizing radiation. They demonstrate that the paradigm, which states that low-dose effects can be predicted high-dose effects, cannot be systematically applied in radiation biology in general and gerontology in particular.

One-year chromaffin cell allograft survival in cancer patients with chronic pain: morphological and functional evidence.

The control of chronic pain through transplantation of chromaffin cells has been reported over the past few years. Analgesic effects are principally due to the production of opioid peptides and catecholamines by chromaffin cells. Clinical trials have been reported with allografts consisting of whole-tissue fragments implanted into the subarachnoid space of the lumbar spinal cord (14,19,36). In the present study, allogeneic grafts were successfully used to control chronic pain in two patients over a period of 1 yr based on patient reported pain scores, morphine intake, and CSF levels of Met-enkephalin. Macroscopic examination at autopsy located the transplanted tissue fragments in the form of multilobulated nodules at the level of the spinal axis and cauda equina. Immunocytochemical microscopy showed neuroendocrine cells are positive for chromagranin A (CGA), and enzymes tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DbetaH). The results suggest that there is a relationship between analgesic effect, Met-enkephalin levels in CSF, and the presence of chromaffin cells surviving in spinal subarachnoid space.

[Excretory azoospermia: contribution of exploratory and curative surgery. Apropos of a follow-up of 33 patients]. / Les azoospermies excrétoires: apport de la chirurgie exploratrice et curative. A propos du suivi de 33 patients.

Thirty-three patients with azoospermia of apparently excretory origin underwent surgery for epididymis-deferens anastomosis and/or epididymal sperm puncture. Pathology examinations of the epididymal fluid and biopsies of the testicles or epididymis were performed at surgery. Based on the clinical presentation, sperm results and per-operative findings, patients were divided into six groups by etiology: idiopathic azoospermia (n = 5), post-infectious azoospermia (n = 15), agenesia of the excretory (n = 6) or secretory (n = 3) ducts, vasectomy (n = 2), and obstruction of the ejaculatory ducts (n = 2). Peroperative identification of spermatozoa at epididymal puncture or biopsy was statistically more frequent in patients with agenesis of the excretory ducts than in patients with post-infectious or idiopathic azoospermia. Biopsies of the testicle led to the diagnosis of secretory azoospermia in 3 cases and revealed a functional parenchyma in all the other groups of patients. Epididymis-deferens anastomosis was performed in 45% of the cases and was successful in 13%. Rate of fertility with the intracytoplasmic sperm injection was 33%; there was no difficulty in using fresh or frozen sperm. Clinical pregnancy was continued to term with frozen sperm. This study confirms that testicular function is preserved in excretory azoospermia. With or without epididymis-deferens anastomosis, epididymal spermatozoa can generally be preserved for later use. Couples should however be counselled on the delays to contraception which may vary from months to years.

[Alveolar lavage cytological data in lung transplants free of bronchial superinfection. Toulouse Lung Transplantation Group]. / Données cytologiques du lavage alvéolaire chez le transplanté pulmonaire indemne de surinfection bronchique. Groupe de Transplantation Pulmonaire de Toulouse.

Total and differential cells counts from 173 bronchoalveolar lavages (BAL) carried out in 19 lung transplanted recipients have been analysed. The patients were separated into seven groups: (a) those without detectable complications (86 BAL); (b) those with acute rejection (AR) (26 BAL); (c) those with bronchiolitis obliterans (BO) (21 BAL); (d) those with intra-alveolar cytomegalovirus (CMV) but asymptomatic (11 BAL); (e) those with an associated intra-alveolar CMV and AR (7 BAL); (f) those with CMV pneumonia (11 BAL); (g) those with infectious pneumonia excluding CMV (11 BAL). The alveolar cytological data were compared to those from control subjects who were smokers (7 BAL) or non-smokers (8 BAL). Excluding the BO group, the total cell count from the BAL of transplanted patients was significantly higher than in the non-smoking controls and was approaching that of the smoking controls. The lymphocytes count was significantly higher in transplanted patients without complication and in the AR group than in the controls, but there was no significant difference between the two groups. There was a significantly greater eosinophilia at the time of AR than either before or after. Eosinophilia was also higher in grade 3 AR than in grade 1 or 2 AR. These data stress the absence of any correlation between the lymphocyte count in the BAL and the existence of AR. Alternatively, the alveolar eosinophilia observed at the time of AR does not allow the possibility of an infection to be elimitated with certainty.

Transplantation of human chromaffin cells for control of intractable cancer pain.

Adrenal medullary chromaffin cells produce high levels of endogenous opioid peptides. Recent data suggest that transplantation injected locally into the spinal subarachnoid space reduced intractable malignant pain. In order to determine the feasibility, the efficacy and the risks of using adrenal medullary tissue for control of irreducible pain, we have developed a transplantation protocol on cancer pain patients selected when they required chronic intrathecal injection of morphine and progressively increasing doses to maintain the level of analgesic effects. At the present time, our clinical trial involves 8 patients. We report here our initial results (mean follow-up: 5 months). The various data collected before and after the intrathecal administration of chromaffin cells included: 1) Pain evaluation over time, with concomitant narcotic intake, 2) CSF sampling through an implanted access port to determine the following biological parameters: biochemical assay for opioid peptides, cell count and phenotyping of lymphocytes, 3) peripheral blood samples for lymphocyte typing. The results confirm the efficacy of adrenal medullary transplantation into spinal CSF for controlling irreducible cancer pain. Complementary intrathecal and oral morphine were totally stopped in 2 cases and stabilized in 5 others. It seems essential to have an important volume of grafted tissue to achieve analgesia with high levels of metenkephalin in CSF. A progressive decrease in metenkephalin release was observed from 2 to 4 months after the transplantation. Two patients with a long-term follow-up (8 and 12 months) needed another intrathecal chromaffin cell graft.

Antibodies to bladder-tumor-associated antigens as prognosis probes in the flow-cytometric analysis.

Monoclonal antibodies directed against bladder tumor cells (10D1, 7C12, 6D1, 3C6, G4 and E7) and human leukocyte antigen (HLe1) were tested by flow cytometry on 68 bladder tumors involving 10 grade I, 29 grade II and 29 grade III tumors (WHO classification). According to their evolution stage, these tumors can be subdivided into 17 stage Pa, 34 stage P1, 7 stage P2 and 10 stage P3. Fifteen normal bladder samples were used as a control. Analysis of DNA content revealed a first group of 31 tumors with a unimodal DNA profile. In the second groups of 37 tumors, the DNA profile was bimodal. Cells from grade I tumors were labelled with 10D1 and 6D1 antibodies; all these cells showed a unimodal DNA profile. Grade III tumor cells were labelled with antibodies G4 and E7; most of these cells showed a bimodal DNA profile. The percentage of HLe1-positive cells decreased with the pathological grade and stage of tumor. The composition of infiltrating leukocytes was different in unimodal and bimodal tumors. In conclusion, cells of low-grade tumors can be identified with 10D1 and 6D1 antibodies, and antigens recognized by G4 and E7 antibodies are mostly expressed by aneuploid cells. HLe1 antibody demonstrates the importance of the inflammatory reaction in bladder tumors. Moreover, in flow cytometry, leukocytes within a tumor could be used as internal reference for precise measurement of the DNA content of tumor cells.

Bronchoalveolar lavage in liquid paraffin pneumonitis.

We evaluated cells and lipids recovered in the bronchoalveolar lavage fluid from seven patients with liquid paraffin pneumonitis. For each patient, the BALF was whitish with oil droplets on the surface. Alveolar macrophages contained numerous, large vacuoles that did not react with May-Grunwald-Giemsa, Papanicolaou, or periodic acid-Schiff but were stained in black with Sudan B, orange with Sudan III and red with oil Red O. Liquid paraffin was identified on thin layer chromatography of BALF-extracted lipids as a very hydrophobic compound migrating on the solvent front as control liquid paraffin. This abnormal spot was definitely identified as liquid paraffin by infrared spectroscopy and gas liquid chromatography for the first patient. The number and percentage of AMs were largely decreased in the BALF of each patient, whereas the number of neutrophils, eosinophils and lymphocytes was increased. These findings suggest that this cell-mediated inflammatory response plays a role in the development of interstitial fibrosis at late stages of liquid paraffin pneumonitis.

Effects of angular speed in responses of Paramecium tetraurelia to hypergravity.

The paper shows the results of investigations carried out in a single cell organism. Paramecium tetraurelia exposed to different gravitational levels. Hypergravity resulted in a decrease in cell growth rate. The responses depend on g level and angular speed of the centrifuge; furthermore they depend also on small short fluctuations in g levels, delta g, due to the swimming of the cells inside the culture tubes. Delta g depends on angular speed and size of the holding device. The inhibitory effect of hypergravity, for the same angular speed, increases with respect of the diameter of the culture tubes.

Flow cytometry analysis of urothelial cell DNA content according to pathological and clinical data on 100 bladder tumors.

DNA content of 100 bladder tumors (34 grade I, 42 grade II and 24 grade III, WHO classification) were studied by flow cytometry. Ten normal bladder samples were used as control. The 100 bladder tumors could then be separated into two groups. A first group of 60 tumors (60%) had a unimodal distribution with a diploid peak and a DNA index close to 1.0, 32 grade I, 22 grade II and 6 grade III tumors displayed this pattern as did the 10 normal bladders. The second group (40%) had a bimodal distribution with two peaks, the first one (diploid peak) with a DNA index of 1.0, the second (aneuploid peak) with a DNA index greater than 1.0. Two grade I, 20 grade II and 18 grade III tumors belonged to this group. Frequency of the aneuploid peak increased with tumor grade and infiltration progression. Hence 6% of grade I, 48% of grade II and 75% of grade III tumors showed an aneuploid peak as well as 8% of Pa, 46% of P1, 73% of P2 and 87.5% of P3 stage tumors. This study showed that a good correlation exists between flow-cytometric, pathological and clinical data.

Influence on cell proliferation of background radiation or exposure to very low, chronic gamma radiation.

Investigations carried out on the protozoan Paramecium tetraurelia and the cyanobacteria Synechococcus lividus, which were shielded against background radiation or exposed to very low doses of gamma radiation, demonstrated that radiation can stimulate the proliferation of these two single-cell organisms. Radiation hormesis depends on internal factors (age of starting cells) and external factors (lighting conditions). The stimulatory effect occurred only in a limited range of doses and disappeared for dose rates higher than 50 mGy/y.

Some ultrastructural aspects of the vitreoretinal juncture.

The ultrastructural study of the vitreoretinal juncture was carried out by comparing two groups of patients: 6 young subjects under 25 years and 6 older subjects over 70. Retinal samples were taken from the posterior pole, the equator and the extreme periphery. Our work shows that the morphological modifications of the vitreoretinal juncture during the aging process vary following location. The most original alteration, never observed before, appears in the equatorial region: it consists of a widening of the intercell space filled with fibrils adhering to Müller's cells. It builds up an adhesion zone between the vitreous and the retina which is not visible during clinical investigations; it can create a tear during the posterior vitreous detachment.

Ultrastructural investigations on two lymphoid cell lines from Niemann-Pick disease type B.

Lymphoid cell lines (LCL) were established by Epstein-Barr Virus (EBV) transformation of blood B-lymphocytes from two different patients affected with Niemann-Pick disease (NPD) type B. Those lines were severely deficient in sphingomyelinase activity (8% and 10% residual activity). Ultrastructural investigations showed in both these lines the presence of numerous osmiophilic, dense and pleiomorphic inclusions characteristic of lysosomal storage (due to the accumulation of amphiphilic lipids) similar to those observed in tissues from NPD.

[Ultrastructural aspects of the dormant Artemia embryo]. / Aspects ultrastructuraux de l'embryon dormant Artemia.

The ultrastructure of the Artemia cyst and larval organism was investigated. The morphology of the blastomere and of the cell organelles was described. Investigations showed the presence of two mitochondria populations: one located in the cytoplasm, the other one embedded inside the yolk platelets. Relations between yolk and mitochondria are more obvious when considering the first steps of embryonic development.