[Neonatal onset of Menkes disease: diagnosis interest of cupremia and microscopic examination of the hairs]. / Maladie de Menkes néonatale: intérêts diagnostiques de la cuprémie et de l'examen microscopique des cheveux.

BACKGROUND: Menkes disease is a rare X-linked disorder due to a defect in intracellular copper transport. Clinical symptoms appear during the first months of life, with a progressive developmental delay leading to death within a few years. Diagnosis is confirmed by the demonstration of copper retention in fibroblasts and/or DNA testing. However, these investigations are complexes and time consuming. CASE REPORT: We report 1 case of Menkes disease with neonatal onset, diagnosed on multiple organ failure, hypothermia, and major central nervous system damage, leading to death in a few weeks. The diagnosis, suggested by the clinical features, was rapidly supported by the microscopic examination of the hairs, showing pili torti, and the demonstration of severely decreased levels of plasma copper and ceruloplasmin. Diagnosis was further confirmed by the demonstration of an increased copper uptake and retention in fibroblasts. CONCLUSION: This report highlights the clinical variability of Menkes disease with the possibility of a neonatal onset. Microscopic examination of the hairs and the determination of copper and ceruloplasmin plasma levels are simple and inexpensive investigations, which can provide rapidly valuable information to support this diagnosis.

Oxidative stress involvement in chemically induced differentiation of K562 cells.

The erythroid differentiation of K562 cells could be achieved by exposure to several pharmacologic agents, including hemin, butyric acid (BA), and anthracycline antitumor drugs such as aclarubicin (ACLA) and doxorubicin (DOX). When used at subtoxic concentrations, these drugs induce the overexpression of erythroid genes, leading to hemoglobinization of cells. Because anthracyclines are known to generate oxidative damage, we intended to demonstrate the involvement of an oxidative stress in the chemically induced differentiation process. The addition of antioxidants to anthracycline- and BA-induced cells decreased their growth and dramatically reduced the percentage of differentiated cells at day 3. Northern blot analysis showed that antioxidants also decrease the expression of erythroid genes and related transcription factors in induced cells. Moreover, analyses of oxidative stress markers showed that treatment with BA, ACLA, and DOX lead to a decrease in reduced glutathione and antioxidant enzymes (glutathione peroxidase [GPx], glutathione reductase [GRase], CuZn superoxide dismutase [SOD], and catalase [CAT]). In addition, DOX increased thiobarbituric acid reactants (TBARs), and MnSOD activity was decreased by BA and DOX. Finally, the production of reactive oxygen species (ROS) by differentiating agents was demonstrated using the dihydroethidium probe in a microspectrofluorometric assay. Altogether, these results strongly suggest the involvement of an oxidative stress generated by BA or anthracyclines as the first step in the irreversible differentiation process. Additionally, these results underline the differences between BA, ACLA, and DOX molecular mechanisms.

Value of nocturnal penile tumescence and rigidity (NPTR) recording in impotent patients with multiple sclerosis.

The etiology of impotence in patients with multiple sclerosis (MS) is difficult to assess due to the possibility of normal nocturnal penile tumescence and rigidity (NPTR) recording despite neurologic involvement. Sixteen patients with MS and impotence were studied with Rigiscan, cavernous artery doppler, neurophysiological tests and intracavernous injection of PGE1. When more restrictive criteria of normality than usual are used for Rigiscan (rigidity > or = 80% and/or duration > 30 minutes), an inverse relationship between NPTR recording and sexuality score or PGE1 dose is reported. No significant difference is noted for neurophysiological tests. With such criteria, Rigiscan alone or combined with intracavernous PGE1 is a valuable means to differentiate neurogenic from psychogenic impotence in MS patients. Neurophysiological tests are of limited clinical value.

Menkes disease: study of the mitochondrial respiratory chain in three cases.

Mitochondrial oxidative metabolism in three patients with typical Menkes disease was studied. In two cases, a general decrease in all of the respiratory chain complex activities (I, II, III and IV) was observed. However, in the most severe case, these activities were entirely normal. Our results emphasize the diversity of the cellular expression of Menkes disease which can, in some cases, be associated with a mitochondrial encephalomyopathy.

Modulation of natural killer cell functional activity in athymic mice by beta-carotene, oestrone and their association.

In athymic mice, Natural Killer (NK) cells influence the take rate and growth of human malignant tissue xenografts. To confirm preliminary results, comparative experiments were conducted to study the effects of beta-carotene, oestrone and their association on the cytolysis mediated by spleen NK cells from athymic mice receiving these different treatments. Target cells consisted of YAC-1 malignant cells. With a 65% increase of cytolysis (ratio effector/target 50:1), beta-carotene induced a significant activation of NK cells (p < 0.002). This effect could be attributed to its antioxidant properties and confirmed by a moderate increase in erythrocyte glutathione peroxidase activity. On the contrary, oestrone resulted in a significant decrease of cytolysis (p < 0.001). In this case, the prooxidant properties of oestrone could explain its effect on NK cells and agree with the increase of intracellular reduced glutathione level observed. When mice received the combination beta-carotene-oestrone, their opposite effects on NK cell activity were counterbalanced, leading to a moderate change of cytolysis.

Effect of oestrone on the natural killer (NK) cell activity, antioxidant status and tumour growth in athymic mice xenografted with human tumours.

Natural killer (NK) cells have been described as being very sensitive to oxidative stress. Thus it has been previously shown that chronic administration of oestrone in drinking water of athymic mice xenografted with a wide variety of human tumours, increases their growth and development. In this study an investigation was made to see whether oestrone supplementation could influence the NK cell activity by changes in the antioxidant defences which result in an oxidative stress and influence the proliferation of tumours. Supplementary oestrone was administered in drinking water of athymic mice xenografted with two different human tumours which lack oestrogen receptors: a bladder carcinoma and a small-cell lung carcinoma. The growth of the urothelial carcinoma was poorly affected by oestrone, but oestrone significantly (p<0.01) increased the proliferation of the small-cell lung carcinoma. The average uterus weight was increased by 62% in oestrone treated mice with no modifications in plasma zinc and selenium status, nor in erythrocyte copper zinc superoxide dismutase level. Nevertheless a slight decrease in erythrocyte glutathione peroxidase activity was noted. Trace elements and antioxidant enzymes in liver homogenates remained unchanged. Oestrone treatment also had no effect on plasma and liver lipid peroxides. The immune response was evaluated by measuring NK activity of splenocytes against 51Cr labelled YAC-I target cells. A 35.5% decrease in the NK activity (p<0.001) was observed after oestrone treatment and may be responsible for graft tolerance. However, the results of these experiments seem to exclude the role of oxidative stress in the modulation of NK activity.

Recurrent pyelonephritis without vesicoureteral reflux: is there a role for an antireflux procedure?

PURPOSE: To evaluate the results of an endoscopic antireflux procedure in women with recurrent acute pyelonephritis and no evidence of vesicoureteral reflux (VUR) on voiding cystograms. PATIENTS AND METHODS: From 1989 to 1999, 603 female patients were hospitalized for acute pyelonephritis with unilateral loin pain, chills, fever, and a positive urine culture. Of these patients, 48 (8%) had recurrent episodes of acute pyelonephritis and underwent a thorough diagnostic work-up including intravenous urography or renal CT scan, cystoscopy, and voiding cystourethrography (VCUG). Vesicoureteral reflux was demonstrated in 21 patients, who were then offered an antireflux procedure, either surgical or endoscopic. Another 27 patients had no reflux on VCUG; in 15 cases, the upper urinary tract was normal, and the ureteral orifices did not show any abnormality on cystoscopy. The other 12 patients in this group with a normal VCUG had one or more abnormal findings normally associated with VUR: renal scarring in five and ureteral duplication in two. Golf-hole ureteral orifices were noted in two patients. The intravesical ureter was short (< 5 mm) in five patients. In spite of the normal VCU, we offered these patients endoscopic treatment of VUR by submeatal injection of Teflon or microparticulate silicone (Macroplastic). The median follow-up before treatment was 4 years (range 1-15.3 years); 0.3 episodes of acute pyelonephritis per patient-month of follow-up were noted. The frequence of preoperative and postoperative episodes of acute pyelonephritis was compared with Wilcoxon's paired analysis. The median postoperative follow-up was 3.9 years (range 1.1 months-10.2 years). RESULTS: There were no significant postoperative complications. One patient had two episodes of acute pyelonephritis during pregnancy. On the whole, 11 patients (91%) were free of recurrent pyelonephritis after treatment. Overall, 0.003 episodes of acute pyelonephritis per patient-month of postoperative follow-up were observed. The result was statistically significant (P < 0.01). CONCLUSION: Recurrent acute pyelonephritis is frequently related to VUR. Intermittent reflux can be difficult to demonstrate on voiding conventional or nuclear cystograms but can be suspected in the presence of ureteral duplication, renal scarring, or abnormal ureteral orifices. Adult patients with recurrent episodes of upper urinary tract infection and normal cystograms should be considered for an endoscopic antireflux procedure in the presence of anatomic abnormalities commonly associated with reflux.

Biodegradation of anthracene and fluoranthene by fungi isolated from an experimental constructed wetland for wastewater treatment.

Pilot-scale constructed wetlands were used to treat water contaminated by polycyclic aromatic hydrocarbons (PAHs), particularly fluoranthene, and the possible role of fungi present in these ecosystems was investigated. A total of 40 fungal species (24 genera) were isolated and identified from samples (gravel and sediments) from a contaminated wetland and a control wetland. All of them were assayed for their ability to remove anthracene (AC) and fluoranthene (FA) from liquid medium. FA was degraded efficiently by 33 species while only 2 species were able to remove AC over 70%. A selection of 10 strains of micromycetes belonging to various taxonomic groups was further investigated for FA and AC degradation, toxicity assays and phenoloxidases (POx) detection. Interesting and not previously reported species were revealed (Absidia cylindrospora, Cladosporium sphaerospermum, and Ulocladium chartarum). They were all able to highly degrade the PAH-model compounds chosen. An interesting inducibility was noted for Ulocladium chartarum. Degradative ability of fungi was not related to their extracellular POx activity. This study may contribute to the improvement of constructed wetlands for water treatment, which may be enriched in efficient fungi.

Beta-carotene enhances natural killer cell activity in athymic mice.

To study the effects of beta-carotene on Natural Killer (NK) cells, we chose athymic mice whose spleens have a higher percentage of NK cells than conventional mice. Preliminary studies conducted with beta-carotene given intraperitoneally to athymic mice xenografted with a small-cell lung carcinoma resulted in a slight but significant antiproliferative effect (unpublished observations). We speculated that such an activity of beta-carotene was related to its immunostimulating properties. NK cell activity in ungrafted athymic mice as influenced by beta-carotene was studied. Mice received beta-carotene intraperitoneally. Splenic NK cells were labelled with monoclonal antibody and numeration was completed by measurement of their functional activity against YAC-1 malignant cells with a 51Cr release assay. In addition, splenic lymphocytes were evaluated for their reduced glutathione (GSH) content. There was a non-significant increase in the number of NK cells in the spleen, however their killing capacity was significantly (p < 0.01) enhanced after beta-carotene treatment. Also the GSH content of splenic lymphocytes was significantly higher in beta-carotene treated mice. Comparison of the average body weights of treated animals and of their respective controls showed that treatment had no adverse effects.

Characterisation of the acoustic cavitation cloud by two laser techniques.

An experimental investigation of the size and volumetric concentration of acoustic cavitation bubbles is presented. The cavitation bubble cloud is generated at 20 kHz by an immersed horn in a rectangular glass vessel containing bi-distilled water. Two laser techniques, laser diffraction and phase Doppler interferometry, are implemented and compared. These two techniques are based on different measuring principles. The laser diffraction technique analyses the light pattern scattered by the bubbles along a line-of-sight of the experimental vessel (spatial average). The phase Doppler technique is based on the analysis of the light scattered from single bubbles passing through a set of interference fringes formed by the intersection of two laser beams: bubble size and velocity distributions are extracted from a great number of single-bubble events (local and temporal average) but only size distributions are discussed here. Difficulties arising in the application of the laser diffraction technique are discussed: in particular, the fact that the acoustic wave disturbs the light scattering patterns even when there are no cavitation bubbles along the measurement volume. As a consequence, a procedure has been developed to correct the raw data in order to get a significant bubble size distribution. After this data treatment has been applied the results from the two measurement techniques show good agreement. Under the emitter surface, the Sauter mean diameter D(3, 2) is approximately 10 microm by phase Doppler measurement and 7.5 microm by laser diffraction measurement at 179 W. Note that the mean measured diameter is much smaller than the resonance diameter predicted by the linear theory (about 280 microm). The influence of the acoustic power is investigated. Axial and radial profiles of mean bubble diameters and void fraction are also presented.

Biodegradation capability of Absidia fusca Linnemann towards environmental pollutants.

The purpose of this work was to study the bioremediation capability of Absidia fusca Linnemann (Zygomycete) towards different classes of xenobiotics (lignin-derived compounds, chloroaromatic compounds, polycyclic aromatic hydrocarbons) the presence of which in contaminated soils, water and sediments poses a significant risk to the environment and human health. Two strains from different origins were compared. One was from an official collection and grown in non-inducing conditions, while the other was isolated during the course of the survey of fungal flora in a polluted soil from Annaba (Algeria). All data were analyzed and results validated via a statistical treatment. We showed the effect of the factors studied (origin of the strain, xenobiotic) but also the interactions between these factors. The strain of A. fusca isolated from a polluted soil was able to efficiently degrade most of the xenobiotics tested, particularly: pentachlorophenol, phenol, catechol, guaiacol and ferulic acid. This property also existed in the other strain but at a very low level.

Degradation of phenolic and chloroaromatic compounds by Coprinus spp.

Three species of Coprinus: C. sp, C. cinereus and C. micaceus were compared on solid media for some aspects of their physiological behaviour and cultural requirements (temperature, pH, substrate). Constitutive extracellular enzymatic activities were also determined. The Coprinus spp. exhibited different physiological and cultural features. Cultures of the 3 Coprinus species in synthetic liquid medium showed an efficient degradation of phenolic lignin model compounds (catechol, ferulic acid, guaiacol, phenol, protocatechuic acid syringic acid and vanillic acid) and pentachloronitrobenzene, while pentachlorophenol was not metabolized after 5 days perhaps because of a strong adsorption on mycelial biomass. It was suggested that phenoloxidases were not necessarily required for the metabolization of these compounds. Coprinus species may share a common degrading system for monomeric phenolic and chloroaromatic compounds.

Removal of herbicides from liquid media by fungi isolated from a contaminated soil.

Fungi were isolated from soil samples corresponding to pesticide-contaminated soil (CS) and noncontaminated soil (NCS) in the Annaba vicinity (Algeria) and identified. The number of isolates obtained from CS and NCS were 263 and 288, respectively. The most frequent species (Aspergillus fumigatus, A. niger, A. terreus, Absidia corymbifera, and Rhizopus microsporus var microsporus) were not sensitive to the pesticides. The growth of the genus Trichoderma was inhibited by the pesticides, while genera Absidia and Fusarium were stimulated. The 53 species isolated were assayed for their ability to remove metribuzin from liquid medium. Only Botrytis cinerea from NCS and Sordaria superba and Absidia fusca from CS removed more than 50% of the compound after 5 d. Metamitron was very resistant. Among the 21 species tested, only Alternaria solani (from NCS), Drechslera australiensis (from CS and NCS), and Absidia fusca (from CS) reduced the concentration in the medium more than 10% (10-16%). Twelve species were grown with linuron, seven of them were inefficient in removing this compound. The two strains of Sordaria macrospora yielded 22 to 25% depletion, while Botrytis cinerea depleted linuron almost completely. Among the 31 species assayed for their ability to eliminate metobromuron, Botrytis cinerea (from CS and NCS) depleted almost completely the chemical from the medium. Rhizopus oryzae and Absidia fusca from CS removed 40 and 47% of the compound, respectively. No systematic relationships were observed between the soil contamination and herbicide elimination capacities of soil fungi. Absidia fusca and Botrytis cinerea were particularly interesting for bioremediation purposes because they were able to transform efficiently three of the four compounds assayed.

[An uncommon tumor of the urinary bladder: the small cell carcinoma]. / Une tumeur vésicale rare: le carcinome à petites cellules.

Primary small cell carcinoma of the urinary bladder is an uncommon tumor, compared to the frequency of urothelial tumors. Fifty percent of cases are combined with a non endocrine carcinomatous component. We report six new cases of this tumor, three of pure, and three associated with an urothelial carcinoma. Diagnosis is easy established by the immunohistochemical study which show the neuro-endocrine differentiation of these aggressive tumors. Pathologist needs to look for a neuro-endocrine part in all bladder cancer, as its presence modify the treatment. Chemotherapy is used in these cancers, due to their high metastatic power. Places of radical surgery and radiotherapy need to be specified.

Effect of zinc supplementation on resistance of cultured human skin fibroblasts toward oxidant stress.

In purified system zinc has been shown to have an antioxidant role. Its effects on the resistance of cultured cells towards oxidative stress in vitro were examined. Diploid human skin fibroblasts were grown for 21 d in culture media (RPMI 1640 containing 15% fetal calf serum) added with different zinc (Zn) concentrations (100, 125, and 150 microM as Zinc chlorur ZnCl2). In comparison, cell controls were grown in standard culture media (6.5 microM Zn). The intracellular zinc levels of treated fibroblasts increased from 3- to 7-fold (2330 +/- 120 ng/mg protein in 150-microM Zn-treated cells versus 331 +/- 21 ng/mg protein in control cells). The intracellular copper increased 3- fold whereas the iron content slightly but not significantly decreased. The index of basal lipid peroxidation measured as thiobarbituric acid reactants (TBARs) of zinc-supplemented cells was lower than that of non zinc supplemented controls (0.89 mumol/g protein in 150 microM Zn-treated cells versus 1.59 mumol/g protein in controls). At these high doses of zinc, fibroblasts expressed lower antioxidant metalloenzymes activities. Diminished TBARs in Zn treated cells tends to support that Zn acts protectively against free radical mediated damage. However when the cells were challenged with extracellular oxidant stresses mediated by hypoxanthine/xanthine oxidase or hydrogen peroxide (H2O2), an increased toxicity in Zn-supplemented cells was observed. When we applied an intracellular oxidative stress as UV-B or UV-A radiation, Zn-treated fibroblasts were more resistant than cells grown in normal medium. If Zn has shown antioxidant effect in some in vitro or in vivo systems our observations clearly demonstrate that this role is not mediated by antioxidant metalloenzymes.

Cultured human skin fibroblasts absorb 65Zn. Optimization of the method and study of the mechanisms involved.

The radioactive isotope 65Zn was used to study the incorporation of zinc by cultured human skin fibroblasts. The development of the method for studying cell uptake of 65Zn in a minimal synthetic medium is presented. Kinetics carried out on control cultures up to 240 min indicated that zinc uptake occurred in three phases, the first being the most rapid. Temperature and pH affect zinc uptake, in favor of an active transport process. In addition, the rate of incorporation is considerably decreased during the first phases after adding potassium cyanide, during the last phases after adding sodium iodoacetate, and during all the phases if dithioerythritol is used. A hypothesis is therefore proposed according to which several types of mechanisms would be involved in zinc uptake by fibroblasts. At least a part of these mechanisms is energy-dependent.

Evaluation of acrodontiolamide, a chlorinated compound produced by Acrodontium salmoneum de Hoog for cytotoxicity and antimicrobial activity.

A new antifungal compound has been isolated from the culture medium of Acrodontium salmoneum de Hoog. Its structure was previously elucidated and was named acrodontiolamide. However, this compound is not characteristically produced by the genus Acrodontium, it is rather a feature of one isolate of A. Salmoneum coming from the soil of the grotto of La Pierre Saint Martin (France). Production, purification, cytotoxicity and antimicrobial activities of acrodontiolamide are described. Concerning microorganisms, inhibitory activity seems to be specifically restricted to phytopathogenic and entomapathogenic fungi. Acrodontiolamide is not cytotoxic to either normal human cultured cells or tumor cells.

Antimicrobial and antitumoral activities of 6-allyl-5,6-dihydro-5-hydroxypyran-2-one, a lactone produced by a new Drechslera species.

Antifungal, antibacterial and antitumoral properties of 6-allyl-5,6-dihydro-5-hydroxypyran-2-one were researched. This compound was isolated from culture medium of a new Drechslera species from the area of the Dead Sea. The product exhibited a large activity spectrum against microorganisms, with interesting IC 50 values close to those obtained with reference compounds (kanamycin and ketoconazole). Antitumoral potentiality was 10 to 58 times less important than with doxorubicin, however IC 50 obtained were below 4 micrograms/ml, which is the threshold value proposed by National Cancer Institute for preliminary screening of active molecules.

[Diagnostic value of recording nocturnal erections]. / Valeur diagnostique de l'enregistrement des érections nocturnes.

The recording of tumescence and rigidity during nocturnal erections is considered to be the reference examination to differentiate between the organic or psychogenic origin of erectile dysfunction. However, its diagnostic value is more doubtful in the absence of normal rigidity values. 170 consecutive patients were submitted to this type of recording (RigiScan, Dacomed, USA). With the exception of patients suffering from multiple sclerosis, all patients with erectile dysfunction considered to be psychogenic obtained a rigidity > or = 60%. 59% of these patients were detected by this method (sensitivity: 59%, specificity: 87%). In contrast, no conclusion can be drawn from a pathological nocturnal recording (32% of psychogenic patients) and finally, in view of the aetiologies of this series, this examination was considered to be useful in 29% of patients.

[Testicular microlithiasis: ultrasonographic diagnosis and semiologic value]. / Microlithiases testiculaires: diagnostic échographique et valeur sémiologique.

Testicular microlithiasis corresponds to calcifications in the lumen of seminiferous tubules. This rare condition, primarily diagnosed histologically, is usually discovered on scrotal ultrasonography. The characteristic appearance combines multiple microscopic hyperechoic areas disseminated bilaterally throughout the testicular parenchyma. The coexistence of microlithiasis and testicular neoplasms has been described and raises the problem of the therapeutic approach following incidental discovery on scrotal ultrasonography performed for other reasons. The authors report a case of testicular microlithiasis discovered on ultrasonography performed after testicular trauma.