Mycoplasma genitalium screening in a specialized French unit: A retrospective study.

OBJECTIVE: Mycoplasma genitalium (MG) infection accounts for 10-35% of non-gonococcal non-chlamydial (NGNC) urethritis. However, given that most people infected with MG do not develop symptoms and that antimicrobial resistance is increasing worldwide, there is no evidence of any benefits of screening asymptomatic individuals. We conducted this study to describe MG screening practices and outcomes at a French Sexually Transmitted Infections (STI) center in which MG testing was performed selectively and multiplex assays were not carried out [i.e., simultaneous screening for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and MG]. METHODS: A retrospective, observational, single-center study was conducted at the STI unit of Saint-Louis Hospital in Paris. The records of all patients undergoing MG testing from January 1st, 2017, to December 31st, 2018, were reviewed. The primary aim of the study was to describe and evaluate the proportion of MG-positive (MG+) patients among those tested. Secondary objectives were determination of the prevalence of MG+ status among symptomatic patients, risk factors associated with MG infection, and therapeutic modalities and efficacy. RESULTS: Two hundred and forty-nine patients underwent MG testing, 28 (11%) of whom were positive (MG+). The prevalence of MG+ status among symptomatic NGNC patients was 12%. HIV-positive (HIV+) status was significantly associated with MG+ status in univariate and multivariate analyses (Odds Ratio=7.3, 95% Confidence Interval 1.3-41.7; P=0.02). Twenty-three patients (85%) received antibiotics. Eighteen (67%) received azithromycin for 5 days, but 7 had clinical resistance. No quinolone resistance was reported. CONCLUSION: Despite unavailability of multiplex testing at our facility, which led to targeted-only screening for MG, its relatively high local prevalence is in keeping with what is generally observed at similar facilities across the world, where use of multiplex tests enables systematic screening for MG alongside NG and CT. This reinforces the current recommendations in Europe, France and the US against systematic MG testing or treatment in asymptomatic patients.

Detection and localization of a Ca2+-ATPase activity in Toxoplasma gondii.

Toxoplasma gondii, the agent causing toxoplasmosis, is an obligate intracellular protozoan parasite. A calcium signal appears to be essential for intracellular transduction during the active process of host cell invasion. We have looked for a Ca2+-transport ATPase in tachyzoites and found Ca2+-ATPase activity (11-22 nmol Pi liberated/mg protein/min) in the tachyzoite membrane fraction. This ATP-dependent activity was stimulated by Ca2+ and Mg2+ ions and by calmodulin, and was inhibited by pump inhibitors (sodium orthovanadate or thapsigargin). We used cytochemistry and X-ray microanalysis of cerium phosphate precipitates and immunolabelling to find the Ca2+, Mg2+-ATPase. It was located mainly in the membrane complex, the conoid, nucleus, secretory organelles (rhoptries, dense granules) and in vesicles with a high calcium concentration. Thus, Toxoplasma gondii possesses Ca2+-pump ATPase (Ca2+, Mg2+-ATPase) as do eukaryotic cells.

Multianalysis of trace elements in mosses with inductively coupled plasma-mass spectrometry.

As a part of an air-pollution biomonitoring survey, a procedure using inductively coupled plasma-mass spectrometry (ICP-MS) and microwave digestion was developed to achieve a high sample throughput and guarantee the accuracy of the results. This article presents an analytical method to measure 22 trace elements. As, Ba, Cd, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mo, Ni, Pb, Rb, Sb, Sr, Th, Tl, U, V, W were analyzed in 563 mosses collected in France. The digestion was performed in polytetrafluoroethylene (PTFE) vessel using the mixture HNO3-H2O2-HF. The data were reprocessed taking into account the drift curve calculated for each element. The detection limits (DL) calculation was based on the standard deviations of the reagent blanks concentrations. The DL varied from one batch to another, because of the heterogeneity of the mosses' elemental contents. The DL ranged between 0.001 microg/g (Cs, Tl) and 70 microg/g (Fe) and were mainly around 0.01 microg/g (As, Cd, Ce, Co, Hg, La, Mo, Sb, Sr, U, V, W). The detection limits obtained were in agreement with the concentrations observed in the samples, except for Hg and Ni. The reproducibility between duplicates and the analytical precision were near 10%. The procedure was tested with the mosses' reference materials.

Effect of hydroxyapatite sintering temperature on intracellular ionic concentrations of monocytes: a TEM-cryo-X-ray microanalysis study.

Hydroxyapatite used as bone replacement can lead to particle release in the implantation site. These particles interact with monocytes, which are the first immune cells to colonize the implant and an inflammatory site. Thanks to cryo-X-ray microanalysis, we can observe cells in a state close to the physiological one and we have access to diffusible ions. We paid particular attention to the potassium-to-sodium ratio, which is one of the best viability criteria. We used this method to study the interaction between three hydroxyapatite particles treated at three different temperatures (not treated, treated at 600 degrees C and 1180 degrees C), and monocytes. In the culture condition, the hydroxyapatite treated at 1180 degrees C underwent the least dissolution. We demonstrate that monocytes were altered by the three hydroxyapatite particles. The hydroxyapatite particules treated at 600 degrees C were found to be more toxic.

X-ray microanalysis of organic thin sections in TEM using an UTW Si(Li) detector: comparison of quantification methods.

We compared Hall's peak to continuum ratio method with a peak ratio method in order to quantify light elements (C, N, and O) in organic specimens as a model for biological thin sections. X-ray spectra were recorded by an energy dispersive X-ray spectrometer equipped with an ultra thin window detector. Spectra were processed by means of a top-hat filter adapted to peak full-width half maximum. The peak intensities were measured by multiple least square fitting to reference spectra. For most elements of biological interest, theoretical and experimental k-factors were determined. Absorption correction was found to be important for quantitation of carbon, nitrogen, and oxygen. Boron was efficiently detected; however, quantitative analysis was not possible. We conclude from our experiments that the peak ratio method is more suitable for quantitation of elemental concentrations in biological thin sections than the peak to continuum method.

Dissolution Kinetics, Selective Leaching, and Interfacial Reactions of a Bioglass Coating Enriched in Alumina.

Bioglass coatings are interesting for developing a direct bond between prostheses and bone. But the high solubility of these materials limits their application. The addition of alumina can be used to control their solubility, but may inhibit the bonding mechanisms. In this paper, we study a bioglass in the SiO(2)-Na(2)O-CaO-P(2)O(5)-K(2)O-Al(2)O(3)-MgO system. After delays of implantation from 2 to 12 months, the bioglass/bone interface is characterized by energy-dispersive X-ray spectroscopy coupled with scanning transmission electron microscopy. Bioglass dissolution can be decomposed into three steps with selective leaching. Results show that, at 2 months after implantation, the bioglass is composed of Al, Si, Ca, and P. Alumina addition increases the coating stability without inhibiting the bonding process. Complex physicochemical reactions take place at the bioglass periphery. The coating bonds to bone through a Ca-P layer on top of a pure Si-rich layer. These phenomena are associated with bioactivity properties, which occur for up to 6 months. After 12 months, the bioglass is composed of silicon. Copyright 2001 Academic Press.

In vitro effects of zirconia and alumina particles on human blood monocyte-derived macrophages: X-ray microanalysis and flow cytometric studies.

The cytocompatibility of two particulate bioceramics, zirconia and alumina, was studied using human blood monocytes driven to differentiate into mature macrophages with granulocyte macrophage-colony-stimulating factor. Changes in individual cell elemental composition, particularly sodium and potassium content, were assessed by X-ray microanalysis of ultrathin freeze-dried sections. Phagocytosis and respiratory burst of macrophages exposed to biomaterial for 7 days were analyzed under flow cytometry using uptake of fluorescent latex beads and 2'7'-dichlorofluorescien diacetate oxidation, respectively. Zirconia and alumina particles were found to decrease the intracellular potassium/sodium ratio (an index of cell vitality) significantly (p<.01) in 7-day-cultured macrophages compared to control cells cultured out of material. Phagocytosis of both ceramic particles by macrophages was followed by a concomitant decrease in cell phagocytic ability (27%) and a marked altered oxidative metabolism (>2 times reduced by zirconia and >5 times reduced by alumina). The present study clearly demonstrates that reduction of the phagocytic capacity of macrophages associated with altered oxidative metabolism caused by biomaterial particles is characterized by changes in intracellular elemental content. Thus, investigation of cellular homeostasis by electron probe microanalysis together with analysis of functional changes may improve estimation of biomaterial cytocompatibility.

Integration of dense HA rods into cortical bone.

HA ceramics are daily used in human surgery for bone healing partly due to their ability to integrate into bone. They are generally used under a macroporous form. The behaviour of dense HA after implantation is not so well known. We implanted within cortical sheep femurs dense pure HA-ceramics cylinders for periods from 2 weeks to 18 months. The samples were then sectioned and examined using back-scattered and secondary SEM and the interface was analysed using EDS. Histomorphometry measurement was also performed using an image analysis device coupled to a light microscope. It appeared that the cylinders were in direct contact with immature bone after three weeks. The bone maturated within three months. The implant surface showed moderate signs of resorption and some grains were released from the surface. The resorption zone was only a few microm thick after 18 months. The bulk ceramic contained default zones of increased porosity. They can constitute fragile zone when located close to the surface in which the resorption rate is increased. We conclude that dense pure HA is poorly degraded when implanted in cortical bone. Degradation depends on the defaults found on the ceramic structure and the remodelling of bone surrounding the material.

Tachyzoite calcium changes during cell invasion by Toxoplasma gondii.

The invasion of host cells by the obligate intracellular protozoan parasite Toxoplasma gondii is calcium dependent. We have identified two calcium storage areas in tachyzoites, the endoplasmic reticulum and vesicles that contain high concentrations of calcium as amorphous calcium phosphate precipitates. Our data indicate that these vesicles slowly lose their calcium during the intracellular development of the tachyzoite as their nucleus phosphorus content increases. We found fluctuations in the sulfur content of the tachyzoite during invasion following the exocytosis of protein from the secretory organelles, with a loss of sodium and chlorine, and the uptake of potassium from the host cell cytoplasm. We demonstrated that penetration of the tachyzoite into the host cell was accompanied by increases in the concentrations of phosphorus and sulfur in the host cell nucleus, probably due to increased transcription. The cytosol sodium concentrations decreased, while the potassium content increased. Thus, the subcellular element distribution of tachyzoites and host cells changes during invasion and intracellular growth of the parasites. In addition, our results indicate that tachyzoite calcium might be involved in the egress of the parasite from the host cell.

X-ray microanalysis of native airway surface liquid collected by cryotechnique.

The airway surface liquid (ASL) that lines the surface epithelium of the tracheobronchial tree is of vital importance to the airway defence against microbial invasion and damage due to environmental factors. Little is known about the ASL collected in situ in native conditions, owing to difficulties in collecting ASL without causing damage to the airway mucosa. We have developed a method to collect and analyse the elemental composition of tracheal ASL in pathogen-free mice. A specially designed cryoprobe, adapted to the internal curvature of the mouse trachea, was used to collect the native ASL from the tracheal surface. The complete ASL elemental composition including [Na] = 5.5 +/- 0.3, [Cl] = 1.3 +/- 0.3, [K] = 1.1 +/- 0.2, [Ca] = 1.2 +/- 0.3, [P] = 1.5 +/- 0.8, [S] = 1.7 +/- 0.4 and [Mg] = 1.3 +/- 0.4 mmol L-1 was determined by X-ray micro-analysis. We demonstrate here that the technique that we used for ASL collection maintained perfectly the airway epithelial integrity and functionality.

X-ray microanalysis of native airway surface liquid collected by cryotechnique.

The airway surface liquid (ASL) that lines the surface epithelium of the tracheobronchial tree is of vital importance to the airway defence against microbial invasion and damage due to environmental factors. Little is known about the ASL collected in situ in native conditions, owing to difficulties in collecting ASL without causing damage to the airway mucosa. We have developed a method to collect and analyse the elemental composition of tracheal ASL in pathogen-free mice. A specially designed cryoprobe, adapted to the internal curvature of the mouse trachea, was used to collect the native ASL from the tracheal surface. The complete ASL elemental composition including [Na] = 5.5 +/- 0.3, [Cl] = 1.3 +/- 0.3, [K] = 1.1 +/- 0.2, [Ca] = 1.2 +/- 0.3, [P] = 1.5 +/- 0.8, [S] = 1.7 +/- 0.4 and [Mg] = 1.3 +/- 0.4 mmol L-1 was determined by X-ray micro analysis. We demonstrate here that the technique that we used for ASL collection maintained perfectly the airway epithelial integrity and functionality.

Involvement of calcium and calmodulin in Toxoplasma gondii tachyzoite invasion.

The tachyzoite of Toxoplasma gondii must successfully invade a host cell before it can replicate. Depletion of the Ca2+ in the external medium (EGTA) reduced tachyzoite invasion, suggesting that the initial tachyzoite-host cell interaction is Ca2+ dependent. The interaction of tachyzoites with host cells was also inhibited by Ca2+ channel blockers (verapamil) and calmodulin antagonists (trifluoperazine, calmidazolium). The calmodulin concentrated at the apical end of the tachyzoite could be involved in cytoskeleton movement and conoid extrusion. Invasion also depends on changes in tachyzoite cytosolic calcium. Depletion of Ca2+ with A23187+EGTA and release of Ca2+ from intratachyzoite pools (nuclear and perinuclear areas) inhibited invasion. In contrast, Ca-ionophore and thapsigargin which increase host cell cytosolic Ca2+, significantly decreased tachyzoite invasion. We therefore suggest that the effect of the drug is significantly different from the localized Ca2+ signal that is produced after parasite attachment to its host cell receptors and leads to its internalization into the host cell.

[Femoropatellar osteoarthritis treated by prosthesis. Retrospective study of 50 implants]. / Traitement de l'arthrose fémoro-patellaire par prothèse sphéro-centrique. Etude rétrospective de 50 implants.

MATERIAL AND METHODS: 51 prosthesis were performed in 47 patients and were studied in order to assess long term results. One patient was lost for follow up. The diagnosis was: primitive arthritis in 48 knees and post traumatic arthritis in 3 knees. 25 knees had patellofemoral subluxation, 16 patients had been previously operated on patellofemoral joint. The average age at the time of surgery was 60.5 years and follow up time averaged 3 years. 22 prosthesis had more than 4 years follow-up. The operations were performed using a lateral approach with tibial tubercle osteotomy and lateral patellar retinaculum release. The trochlear component was asymetric and made of chrome cobalt alloy; the patellar component was shaped in polyethylene. Both components were cemented. RESULTS: Using the Guepar scoring system, 41 (82 per cent) PFA were rated excellent or good and 9 poor. Post operative roentgenograms demonstrated patellofemoral alignment in all knees, even in cases of preoperative patellar subluxation, without tilting of the patella. Follow up roentgenograms demonstrated progressive but moderate deterioration of the tibio femoral joint in 9 cases. In 3 cases, more severe tibio femoral arthritis were treated by T.K.R one to three years after P.F.A. In one of these knees, we discovered an asymptomatic loosening of trochlear component. The last 6 poor results were always due to persistent pain. DISCUSSION: The purpose of this study was to assess the results of P.F.A. and to clarify the indications. It appears that the 82 per cent success rate of this study may be compared with the results of literature. All the authors have found P.F.A. to be a viable solution, preferable to patellectomy (even in older patients) or isolated patellofemoral realignment. The clinical and radiological results did not deteriorate with time. Persistent patellofemoral malalignment was not observed in this study thanks to the use of a lateral approach. The presence of tibio femoral arthritis adversely affected the outcome, but accelerated changes on the tibio femoral joint after P.F.A. were not observed.

A single photon emission computed tomography study of cerebral regional perfusion changes induced by a learning task in subjects with mild memory impairment.

Fifteen nondemented subjects with memory complaints underwent serial single photon emission computed tomography (SPECT) studies with technetium-99m-d, l-hexamethyl-propylene amine oxime (HMPAO) as tracer. Scans were carried out under a baseline conditions and during the learning phase of the Memory Efficiency Profile (MEP), a combined visual perception and memory task developed by Rey. Results indicate a positive correlation between activation, as indexed by HMPAO uptake, and neuropsychological assessment. Right temporal activation was correlated with MEP immediate recall. The right cerebellum was correlated with both MEP immediate and delayed recall. This study suggests that SPECT can show cortical activation during cognitive performance in subjects with mild memory impairment.

Subcellular calcium localization in Toxoplasma gondii by electron microscopy and by X-ray and electron energy loss spectroscopies.

The localization of calcium in Toxoplasma gondii tachyzoites was studied at the ultrastructural level, with a cytochemical pyroantimonate precipitation method (PA) and controlled by EGTA chelating and EDX and EELS microanalyses. Appropriate conditions for material preparation, fixation and embedding, were defined. The proportion of precipitates that were either free or inside vacuoles and their distribution inside Toxoplasma appeared to be PA dose-dependent. Precipitation mainly occurred in the anterior pole of the Epon-embedded tachyzoites. EDX and EELS analyses showed that out of 30 PA precipitates inside tachyzoites, 78% contained Ca. In Melamine sections, 96% of the tachyzoites had intracellular precipitates and the membrane complex was stained; 25% of the tachyzoites inside host cells contained PA-Ca precipitates, but most of them were retained in the reticular network of the parasitophorous vacuole. Melamine embedding appeared to improve the preservation of calcium pyroantimonate precipitates.

Toxoplasma gondii: immunocytochemistry of four immunodominant antigens with monoclonal antibodies.

Four monoclonal antibodies in which diagnostic usefulness has been observed, concerning congenital, acquired, and reactivated toxoplasmosis, were raised against Toxoplasma gondii tachyzoïtes in order to localize immunodominant antigens. On immunoblots, it appears that McAb IV47, McAB GII9, McAb II38, and McAb IE10 identify families of proteins with estimated molecular weights of 28-30 kDa, 30 kDa, 45-50 kDa, and 66-70 kDa, respectively. By immunogold preembedding techniques one can observe an homogeneous labeling of the outer pellicle of the tachyzoïtes with the McAb GII9 and IV47 and a light labeling with the McAb II38 and IE10. The three-dimensional observation of cell surface antigens is performed by applying a modified metal extraction replica method, i.e., A plasma polymerization method of glow discharge by Tanaka (1979). By immunogold preembedding techniques [with saponin permeabilization (0.1%)], and by immunogold postembedding techniques, a labeling of the rhoptries is observed with McAb GII9 and McAb IV47 but essentially all label is found with McAb II38 and IE10. With McAb GII9 a uniform labeling is observed on the cell surface. By immunoenzymatic techniques (peroxidase) a cell surface labeling is observed with the four McAb. Intracellular Toxoplasma, the outer pellicle, and the vesicles of the network (elaborated by Toxoplasma in parasitophorous vacuole) are also labeled with McAb IE10. These results indicate that McAb GII9 recognizes antigens of the antigen family (P 30) located on the cell surface and in the rhoptries. The antigen recognized by McAb IV47 is essentially located on and beneath the Toxoplasma cell surface membrane, and McAb II38 and IE10 identify preferentially rhoptry proteins.

[Treatment of Crohn's disease with azathioprine or 6-mercaptopurine. Retrospective study of 126 cases]. / Traitement de la maladie de Crohn par l'azathioprine ou la 6-mercaptopurine. Etude rétrospective chez 126 malades.

Over the past 15 years (1974-1989), 126 patients with Crohn's disease received azathioprine (n = 123) or 6-mercaptopurine (n = 3). Seven patients were lost of follow-up during the first month and were not analyzed. Among the 119 patients analyzed (52 men, 67 women, mean age 31.6 years), the median duration of treatment was 9.1 months (range: 8 days to 15 years). Most of the 109 patients with active disease at the beginning of the immunosuppressive therapy were either steroid-dependent (n = 63) or steroid-resistant (n = 23); 19 had frequent relapses and/or extensive involvement of the gastrointestinal tract; 4 had severe perianal disease. Among these 109 patients, 25.4, 51, 60.4 and 64.4 percent were in clinical remission at 3, 6, 9 and 12 months (life-table analysis) respectively. There was no difference in outcome of patients with colonic, ileocolonic or small intestinal involvement. Steroid-resistant patients fared better than steroid-dependent patients (81 percent vs 59 percent of remission within the first year; p less than 0.001). In the 62 patients with quiescent disease treated with azathioprine or 6-mercaptopurine, previous remission had been achieved with immunosuppressive therapy in 52, 8 were treated after a bowel resection which was assumed to be curative; 2 had achieved remission after total parenteral nutrition. In these patients, the percentages of relapse were 15.3 at the end of the first year and 20.3 at the end of the second year. Among the 8 patients treated with azathioprine after bowel resection, only one relapse occurred 1.5 months after surgery. Nineteen percent of the patients had adverse reactions that required discontinuation of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)