Phthalate metabolites in the European eel (Anguilla anguilla) from Mediterranean coastal lagoons.

The levels and fate of phthalate metabolites have been poorly evaluated in fish, despite their potential ecotoxicological impacts. The present study aims to characterize the levels of phthalate metabolites in muscle tissue of yellow eels (Anguilla anguilla) from two coastal Mediterranean lagoons, during three sampling periods. Nine phthalate metabolites were detected in >70% of the samples. Slightly higher levels of phthalate metabolites were detected in March and June compared to October, suggesting possible seasonal variations in environmental release and/or phthalate metabolization process by eels. The large sample size (N=117) made it possible to explore correlations between phthalate metabolites' levels and individual parameters, such as body length, age, body condition and hepatic histo-pathologies. Body length and estimated age poorly correlated with phthalate metabolites, suggesting that eels did not accumulate phthalates during growth, contrary to persistent compounds. Eels presented different grades of hepatic fibrosis and lipidosis. A negative correlation was found between the severity of these pathologies in the liver and the sum of phthalate metabolites levels, supporting the hypothesis that eels with damaged liver are less able to metabolize xenobiotics.

Determination of phthalates and their by-products in tissues of roach (Rutilus rutilus) from the Orge river (France).

Seven phthalate by-products were investigated for the first time, in target tissues of roach from a contaminated river of the Ile-de-France district. All parent phthalates were bioaccumulated in liver and muscle and liver contents were correlated with river concentrations (p < 0.01). All metabolites were found in liver, plasma and bile. The mono-iso-butyl phthalate (MiBP; 1.6 µg g(-1) dw of liver), followed by mono-n-butyl phthalate (MnBP; 1.5 µg g(-1) dw of liver) were the most abundant ones. Among the three metabolites of di-ethylhexyl phthalate (DEHP), mono (2-ethylhexyl) phthalate (MEHP) predominated in bile (15.5 ng ml(-1)) and liver (0.237 µg g(-1) dw), whereas in plasma, it was mono (2-ethyl-5-hydroxyhexyl) phthalate - MEHHP (214 ng ml(-1)). In liver, MEHP/DEHP ratios ranged from 0.04 to 0.2. Among the oxidized metabolites, only mono (2-ethyl-5-oxohexyl) phthalate (MEOHP) was correlated (p < 0.05) with parent DEHP and appeared to be a more reliable marker of DEHP impact than the monoester.

Persistent organic pollutants in benthic and pelagic organisms off Adélie Land, Antarctica.

The concentrations of polychlorinated biphenyls (PCB), hexachlorobenzene (HCB), pentachlorobenzene (PeCB) and polybrominated diphenylethers (PBDE) were described in benthic and pelagic species collected off Adélie Land, Antarctica. Strong differences were observed among species, with reduced PeCB and HCB levels in benthic species, and elevated PCB levels in the Antarctic yellowbelly rockcod, the Antarctic sea urchin and the snow petrel. Lower-chlorinated congeners were predominant in krill; penta-PCBs in benthic organisms; hexa- and hepta-PCBs in seabirds and cryopelagic fish. This segregation may result from sedimentation process, specific accumulation and excretion, and/or biotransformation processes. The presence of PBDEs in Antarctic coastal organisms may originate from atmospheric transport and partly from a contamination by local sources. Although POP levels in Antarctic marine organisms were substantially lower than in Arctic and temperate organisms, very little is known about their toxic effects on these cold-adapted species, with high degree of endemism.

[Lymphedema of the lower limbs: CT staging]. / Lymphoedème des membres inférieurs: apport du scanner.

Routinely performed, CT is useful and reliable for staging lower limb lymphedema. We describe methods we utilized. We found in frequency order: skin thickening, subcutaneous tissues area increase in regard the safe limb, perimuscular aponevrosis thickening, fat infiltration: lines parallel to the skin, edematous areas along perimuscular aponevrosis, lines perpendicular to the skin. The lowest fat density is increased on the pathologic side. Subfascial compartment is slightly fattened. We found huge differences between primary and secondary lymphedema for the thigh. Same images may be generated by old or young lymphedema. Rarely useful for positive diagnosis, CT is indispensable for secondary lymphedema staging (initial staging or after a recent increase). It seems us indispensable for any pretherapeutic staging (whole objectively disorders, exact upper limit, infraclinic bilaterality).

[Lymphedema and quality of life]. / Lymphoedème et qualité de vie.

Quality of life measurement shows interesting parameters in chronic lymphedema, related to the illness consequences and therapeutic strains. The ULL27 specific scale is an evaluation instrument which adds to the information about the patient and enables adaptation of the therapeutic strategy and also measurement of the treatment's impact of lymphedema.

The myelin basic protein gene is expressed in differentiated blood cell lineages and in hemopoietic progenitors.

Myelin basic proteins (MBP) are major constituents of the myelin sheath of oligodendrocytes and Schwann cells in the central nervous system and the peripheral nervous system, respectively. We previously showed that MBP-related transcripts are present in the bone marrow and the immune system. These mRNAs are transcribed from a region called 0', consisting of three exons, located upstream of the classical MBP exons; these three exons belong to the long MBP gene otherwise called "Golli-MBP." The most abundant of these mRNAs, now called HMBP (hemopoietic MBP), encompasses the sequence encoded by the region 0' plus exon 1 and part of intron 1 of the classic MBP gene. Antisera to recombinant HMBP proteins are immunoreactive with proteins of about 26-28 kDa in brain, thymus, and spleen. This report demonstrates that HMBP proteins are present in the vast majority (>95%) of thymic T cells, which express the corresponding transcripts, as do mature T cells from lymph nodes and spleen. HMBP mRNAs and proteins are also manifest in the majority of spleen B lymphocytes and in B cell lines. In addition to lymphoid cells, HMBP proteins are in all types of myeloid lineage cells, i.e., macrophages, dendritic cells, and granulocytes, as well as in megakaryocytes and erythroblasts. Finally, HMBP proteins are present in CD34+ bone marrow cells, and, furthermore, in highly proliferative cultures, these CD34+ cells express HMBP RNAs and proteins. Thus, MBP gene products are present both in the nervous system and in the entire hemopoietic system.

[Microglia: origin and development]. / Microglie: origine et développement.

As suggested by Del Rio Ortega a long time ago, it is now widely accepted that microglia are the resident macrophages of the central nervous system. Microglia represent about 10% of the adult brain cell population. We have previously shown that the late embryonic and adult mouse brain contain potential microglial progenitors. We report here that microglial progenitors can be detected in neural folds from embryonic day 8. They originate from the yolk sac in which macrophage progenitors are found from embryonic day 7. We also report that the bulk of microglial cells (about 95%) appear during post-natal development. A major finding is that microglia arise by an intense in situ proliferation comparable to that of neural cells. Taken together, these results show that adult mouse microglia originate from cells migrating from the yolk sac and whose progeny actively proliferates in the brain during development.

Expression of gamma-glutamyl transpeptidase in mouse perivascular astrocytes and in a protoplasmic-like astroglial cell clone.

Gamma-glutamyl transpeptidase (GGT) is known to be present in the central nervous system (CNS) but its cellular localization is still subject to controversy. In this report, we have investigated, with a specific antiserum, the immunolabelling pattern of GGT in the adult mouse CNS at the light and electron microscopic (EM) levels. At the optical level, GGT immunoreactivity ensheathes the majority of vessels in the grey matter. Immunoelectron microscopy shows that labelling is essentially due to the presence of GGT in the astrocytic endfeet which surround vessels. In addition, some pericytes and periendothelial cells are also clearly labelled. We then investigated GGT activity in astroglial cell clones which may represent the in vitro counterpart of the main astroglial cell types. The striking result is that a protoplasmic-like astroglial cell clone shows a noticeable GGT activity, while, in contrast, no activity was detected in the fibrous and the Golgi-Bergmann-like astroglial clones. Taken together, these data indicate that, in the mouse CNS, GGT is essentially present in protoplasmic astrocytes.

[Results of a neurophysiologic consultation in patients with secondary lymphedema of the arm after breast cancer associated with neurological symptoms]. / Résultats d'une consultation neurophysiologique chez des patientes atteintes d'un lymphoedème secondaire du membre supérieur après cancer du sein et présentant des troubles neurologiques.

The aim of this study was to identify the nature of the nerve lesions found in patients with neurological complains and with lymphedema occurring after breast cancer treated by surgery and radiotherapy. Twelve patients treated in a specialised centre for lymphology had clinical and neurophysiological examinations. This study found 9 radiation-induced plexopathies, 1 neoplasic plexopathy, 2 carpal tunnel syndromes (one isolated), 1 cervical root disease and 1 normal examination. The radiation-induced plexopathy is characterised by a chronic neurogenic involvement and the presence of (motor and sensory) proximal persistent conduction blocks. This study demonstrated that it is essential to identify with electrodiagnosis the exact nature of the nerve lesion to determine the most appropriate and effective treatment.

Microglia derive from progenitors, originating from the yolk sac, and which proliferate in the brain.

Microglia, the resident CNS macrophages, represent about 10% of the adult brain cell population. Although described a long time ago, their origin and developmental lineage is still debated. While del Rio-Hortega suggested that microglia originate from meningeal macrophages penetrating the brain during embryonic development, many authors claim that brain parenchymal microglia derive from circulating blood monocytes originating from bone marrow. We have previously reported that the late embryonic and adult mouse brain parenchyma contains potential microglial progenitors [F. Alliot, E. Lecain, B. Grima, B. Pessac, Microglial progenitors with a high proliferative capacity in the embryonic and the adult mouse brain, Proc. Natl. Acad. Sci. U.S.A. 88 (1991) 1541-1545]. We now report that they can be detected in the brain rudiment from embryonic day 8, after their appearance in the yolk sac and that their number increases until late gestation. We also show that microglia appear during embryonic development and that their number increases steadily during the first two postnatal weeks, when about 95% of microglia are born. Finally, the main finding of this study is that microglia is the result of in situ proliferation, as shown by the high proportion of parenchymal microglial cells that express PCNA, a marker of cell multiplication, in embryonic and postnatal brain. Taken together, our data support the hypothesis that terminally differentiated brain parenchymal microglia are derived from cells originating from the yolk sac whose progeny actively proliferates in situ during development.

Pericytes and periendothelial cells of brain parenchyma vessels co-express aminopeptidase N, aminopeptidase A, and nestin.

Within the parenchyma of the CNS, the endothelium of all vessels is surrounded by a layer of cells, pericytes in capillaries and periendothelial or intima smooth muscle cells in other vessels. The origin of these cell types, their relationship, and their role are unclear. However, it has been recently shown that genetically engineered mice that lack pericytes develop microaneurysms at late gestation and die before birth (Lindahl et al. [1997] Science 277:242-245). The goal of this study was to identify in situ molecular markers that would be common to pericytes and periendothelial cells of adult mouse brain. Immunocytochemistry experiments were carried out at the optical and electron-microscopic levels on mouse brain sections with antibodies specific for aminopeptidase N, aminopeptidase A, and the intermediate filament nestin. The results of our experiments show that in all brain parenchyma vessels of all sizes, pericytes and periendothelial cells are immunoreactive for aminopeptidase N, essentially at the plasma membrane level, and are also labeled by nestin specific antibodies, which decorate typical intermediate filaments. In addition, brain pericytes and periendothelial cells are also immunoreactive to monoclonal antibodies to aminopeptidase A. In contrast, pericytes and periendothelial cells do not express microglial markers. Taken together these data show that pericytes and periendothelial intima smooth muscle cells share common markers, suggesting a common origin or function, and are distinct from microglia.

Brain parenchyma vessels and the angiotensin system.

It is now recognized that the brain contains an autonomous angiotensin (AG) system, including the aminopeptidases A and N required for angiotensin metabolism. Using immunohistochemical techniques, we show that capillary pericytes and periendothelial cells of other vessels express aminopeptidase A (APA) and aminopeptidase N (APN) at their plasma membrane in adult mouse brain parenchyma. We therefore investigated the localization of angiotensin II(III), known as putative substrates for these enzymes, as well as that of their precursor angiotensin I. We report here the presence of immunoreactivity to angiotensin I and II(III) around most brain vessels. Angiotensins are present at the plasma membrane of brain parenchymal cells, presumably perivascular astrocytes which are also immunoreactive to AT1-receptor antibodies. The very close relationship between AGII(III) and their metabolizing enzymes APA and APN suggests a specific functional role for brain perivascular angiotensins.

[Computerized tomography of 150 cases of lymphedema of the leg]. / Tomodensitométrie de 150 cas de lymphoedèmes des membres inférieurs.

The aim of this work was to evaluate the usefulness of CT imaging to stage lower limb lymphedemas. Between 1992 and 1997, we studied 150 cases of lymphedema, half idiopathic and half secondary. Methods used are described. In decreasing order of frequency, we found: skin thickening, increased subcutaneous tissue surface area compared with the healthy limb, thickening of the perimuscular aponevrosis, fat infiltration: lines parallel to the skin (parallel), edematous areas along the perimuscular aponevrosis, lines perpendicular to the skin (perpendicular). The lowest fat density was increased on the diseased side. The subfascial tissue showed some fat accumulation. These results were compared with findings reported in the literature. There were very major differences between idiopathic lymphedema and secondary lymphedema of the thigh. Similar images were generally generated by new and long-standing lymphedema. Rarely useful for positive diagnosis, CT is indispensable for establishing stage initially or after recent increase and, in our opinion, is essential for pretherapeutic assessment. The CT-scan gives objective evidence of overall disorders, the exact upper limit of the lymphedema, and sometimes reveals infraclinical bilateral involvement.

Ly-6C is expressed in brain vessels endothelial cells but not in microglia of the mouse.

The Ly-6C antigen is expressed in various cell types of the immune system, including macrophages. Using the monoclonal antibody ER-MP20 which specifically recognises Ly-6C, we have investigated whether brain parenchymatous microglia express Ly-6C in vivo as well as in vitro. In brain sections from developing and adult C57/BI mice, all vessels were strongly immunolabelled. Electron microscopic immunohistochemistry showed that the endothelial cells are the cell type expressing Ly-6C. In contrast, we never observed immunoreactivity on microglia; however, microglial cells proliferating in vitro were strongly ER-MP20 positive. These data show that Ly-6C is not a marker for microglia in vivo.

Lymphedematous skin and subcutis: in vivo high resolution magnetic resonance imaging evaluation.

Physico-chemical and morphologic parameters of skin layers and subcutaneous tissue in lymphedematous limb were studied in vivo using magnetic resonance imaging. High resolution images were obtained with a depth resolution of about 70 microm, using a specific surface gradient coil specially designed for skin imaging and connected to a standard whole-body imager at 1.5 T. Twenty-one patients with unilateral lower extremity lymphedema (11 primary and 10 secondary) were examined. Skin thickness, relaxation times, and relative proton density were calculated in lymphedematous limbs and in contralateral extremities. In diseased limbs, the average skin thickness (2.17 mm) was significantly larger (p = 1.5 x 10(-4)) than that of contralateral limb (1.14 mm). Major cutaneous alterations due to lymphedema took place in dermis. In lymphedematous dermis, the significant increase of relaxation time values could be due to a shift in the equilibrium of water inside this tissue in relation to the interactions between macromolecules and water molecules. In lymphedematous epidermis our results showed an increase in the number of free water protons. Information about water and fat distribution in lymphedema was also obtained using chemical shift weighted images. Our results demonstrated a water retention diffusely spread over the entire dermis, and an important fluid retention located in the interlobular spacing and beside the superficial fascia. Inside the subcutis, the mean thickness of the superficial fat lobules was increased more than that of the deep fat lobules. From all the various measurements we could not distinguish primary from secondary lymphedema.

Transaxial computer tomography of lower extremity lymphedema.

We examined retrospectively 11 patients with isolated unilateral lower limb lymphedema (clinical criteria confirmed by isotope lymphography) using computer tomography. In conjunction with earlier observations, the findings of soft tissue stranding, skin thickening, fat deposition in the epifascial compartment and perimuscular fascial thickening and edema relate to lymph stasis. This noninvasive and relatively simple imaging technique allows analysis of soft tissue changes in leg lymphedema and can be used to evaluate lymphatic insufficiency and its extent as well as document the response to treatment.

The Lck tyrosine kinase is expressed in brain neurons.

The lck gene product, p56lck, is a member of the src-related family of protein tyrosine kinases. It is known as lymphocyte specific and involved in thymocyte development and in the immune response mediated by the T cell receptor. We report that the lck gene is also expressed in adult mouse CNS and that brain p56lck is similar to the thymus protein. In situ hybridization and immunohistochemistry show that the lck gene is expressed in neurons throughout the brain in distinct regions, including hippocampus and cerebellum. In primary cultures from fetal mouse brain, neuronal cells are immunoreactive to Lck antiserum. This suggests that the lck gene product might be involved in a new signal transduction pathway in mouse brain.

A spontaneously immortalized mouse microglial cell line expressing CD4.

We have derived a microglial clone, named C8-B4, from the 8-day mouse cerebellum organ culture which gave rise to distinct astroglial cell lines as previously reported. Indeed, the C8-B4 clone expresses classical microglial markers (MAC1, F4/80, 2-4G2) and appears to be derived from a committed microglial precursor since it does not express differentiation antigens present during the early stage of the monocytic lineage. This microglial clone expresses two characteristics not previously reported for microglial cell lines: it synthesizes the CD4 molecule and produces and releases large amounts of glutamate.

Treatment of secondary lymphedema of the upper limb with CYCLO 3 FORT.

Fifty seven patients with secondary lymphedema of the upper limb after previous treatment for breast cancer were treated for 3 months with an extract of Ruscus + Hesperidin Methyl Chalcone (CYCLO 3 FORT) or placebo according to a double-blind protocol in the context of a controlled clinical trial. All patients also underwent manual lymphatic drainage twice a week for at least one month. With CYCLO 3 FORT, the reduction in volume of arm edema, the main assessment criteria, was 12.9% after 3 months of treatment as compared with a placebo (p=0.009). Decreased edema tended to be more marked in the forearm compared with the upper arm where excess fat deposition seemed to dominate over excess fluid accumulation. CYCLO 3 FORT was well tolerated with minimal adverse reaction.

CD4 expression in neurons of the central nervous system.

CD4 is a member of the Ig gene super family expressed on the surface of many thymocytes and of a subset of T lymphocytes. Human CD4 is the receptor for HIV envelope glycoprotein gp120. Human and mouse CD4 transcripts are expressed in human and mouse central nervous system (CNS), but no corresponding proteins have been reported yet. We have analyzed mRNA expression and carried out immunological experiments on adult mouse brain with probes specific for the long and short CD4 transcripts and with antibodies monospecific for mouse CD4. The main result of these experiments is that the full length CD4 transcript and the CD4 protein are expressed coordinately in neurons throughout the adult mouse brain. CD4 immunoreactivity is also present in brain small vessel walls, ependymal cells, and choroid plexus. The brain mouse CD4 protein is indistinguishable from the thymus protein. In addition, we show that neuronal cells in primary cultures from human fetal CNS are immunoreactive to human CD4 mAbs.