Extracellular matrix educates an immunoregulatory tumor macrophage phenotype found in ovarian cancer metastasis.

Recent studies have shown that the tumor extracellular matrix (ECM) associates with immunosuppression, and that targeting the ECM can improve immune infiltration and responsiveness to immunotherapy. A question that remains unresolved is whether the ECM directly educates the immune phenotypes seen in tumors. Here, we identify a tumor-associated macrophage (TAM) population associated with poor prognosis, interruption of the cancer immunity cycle, and tumor ECM composition. To investigate whether the ECM was capable of generating this TAM phenotype, we developed a decellularized tissue model that retains the native ECM architecture and composition. Macrophages cultured on decellularized ovarian metastasis shared transcriptional profiles with the TAMs found in human tissue. ECM-educated macrophages have a tissue-remodeling and immunoregulatory phenotype, inducing altered T cell marker expression and proliferation. We conclude that the tumor ECM directly educates this macrophage population found in cancer tissues. Therefore, current and emerging cancer therapies that target the tumor ECM may be tailored to improve macrophage phenotype and their downstream regulation of immunity.

Neutrophil gelatinase-associated lipocalin in dogs with chronic kidney disease, carcinoma, lymphoma and endotoxaemia.

OBJECTIVES: To measure serum and urine neutrophil gelatinase-associated lipocalin (NGAL) concentrations in healthy dogs and dogs with chronic kidney disease, neoplasia and endotoxaemia. METHODS: Serum and urine NGAL concentrations were measured in 42 healthy dogs, 11 dogs with chronic kidney disease, 12 dogs with carcinoma, 20 dogs with lymphoma and 15 dogs with lipopolysaccharide-induced endotoxaemia. In dogs with chronic kidney disease, NGAL was measured 3 and 6 months later. RESULTS: Compared with healthy controls, dogs with chronic kidney disease (PÄ0·0008), carcinoma (PÄ0·0072) and lymphoma (PÄ0·0008) had elevated serum and urine NGAL and urine NGAL-to-creatinine ratio. Serum and urine NGAL was not significantly different between dogs with chronic kidney disease, carcinoma or lymphoma (Pê0·12). In dogs with non-progressive chronic kidney disease, NGAL concentrations did not change significantly over the 6-month study period. CLINICAL SIGNIFICANCE: NGAL can be elevated by chronic kidney disease and neoplasia, compared with healthy controls. Further research is needed to determine if uNGAL or uNGAL-to-creatinine ratio is more specific than serum levels to detect chronic kidney disease.

Effect of synthetic colloid administration on coagulation in healthy dogs and dogs with systemic inflammation.

BACKGROUND: Synthetic colloids are often used during fluid resuscitation and affect coagulation. OBJECTIVE: To compare the effects of an isotonic crystalloid and synthetic colloid on coagulation in healthy dogs and dogs with systemic inflammation. ANIMALS: Sixteen adult purpose-bred Beagles. METHODS: Randomized, placebo-controlled, blinded study. Dogs were randomized into one of two groups receiving fluid resuscitation with either 40 mL/kg IV 0.9% NaCl or tetrastarch after administration of lipopolysaccharide or an equal volume of placebo. After a 14-day washout period, the study was repeated such that dogs received the opposite treatment (LPS or placebo) but the same resuscitation fluid. Blood samples were collected at 0, 1, 2, 4, and 24 hours for measurement of coagulation variables. RESULTS: Administration of either fluid to healthy dogs and dogs with systemic inflammation resulted in similar increases in prothrombin time and activated clotting time. In comparison to saline administration, tetrastarch administration resulted in significantly decreased R (P = .017) in healthy dogs, as well as significantly increased activated partial thromboplastin time (P ≤ .016), CL30% (P ≤ .016), and K (P < .001) and significantly decreased platelet count (P = .019), α (P ≤ .001), MA (P < .001), and von Willebrand factor antigen (P < .001) and collagen binding activity (P ≤ .003) in both healthy dogs and dogs with systemic inflammation. CONCLUSIONS AND CLINICAL IMPORTANCE: Tetrastarch bolus administration to dogs with systemic inflammation resulted in a transient hypocoagulability characterized by a prolonged activated partial thromboplastin time, decreased clot formation speed and clot strength, and acquired type 1 von Willebrand disease.

Current Zika virus epidemiology and recent epidemics.

The Zika virus (ZIKV) is a mosquito-borne flavivirus (Aedes), similar to other arboviruses, first identified in Uganda in 1947. Few human cases were reported until 2007, when a Zika outbreak occurred in Yap, Micronesia, even though ZIKV activity had been reported in Africa and in Asia through virological surveillance and entomological studies. French Polynesia has recorded a large outbreak since October 2013. A great number of cases and some with neurological and autoimmune complications have been reported in a context of concurrent circulation of dengue viruses. The clinical presentation is a "dengue-like syndrome". Until the epidemic in French Polynesia, no severe ZIKV disease had been described so far. The diagnosis is confirmed by viral genome detection by genomic amplification (RT- PCR) and viral isolation. These two large outbreaks occurred in a previously unaffected area in less than a decade. They should raise awareness as to the potential for ZIKV to spread especially since this emergent disease is not well known and that some questions remain on potential reservoirs and transmission modes as well as on clinical presentations and complications. ZIKV has the potential to spread to new areas where the Aedes mosquito vector is present and could be a risk for Southern Europe. Strategies for the prevention and control of ZIKV disease should include the use of insect repellent and mosquito vector eradication.

Automated radiosynthesis of N-succinimidyl 3-(di-tert-butyl[(18)F]fluorosilyl)benzoate ([(18)F]SiFB) for peptides and proteins radiolabeling for positron emission tomography.

Recently, silicon fluoride building blocks (SiFA) have emerged as valuable and promising tools to overcome challenges in the labeling of peptides and proteins for positron emission tomography (PET). Herein, we report a fully automated synthesis of N-succinimidyl 3-(di-tert-butyl[(18)F]fluorosilyl)benzoate ([(18)F]SiFB) by a commercially available Scintomics Hot Box 3 synthesis module, to be used as a prosthetic group for peptide and protein labeling. The drying of K2.2.2./K (18)F complex was performed according to the Munich method modified by our group (avoiding azeotropic drying) using oxalic acid to neutralize the base from the (18)F(-) containing QMA eluent. This K2.2.2./K (18)F complex was then used for SiFA (18)F-(19)F isotopic exchange followed by a fast purification by a solid-phase-extraction (SPE) to afford [(18)F]SiFB with an average preparative radiochemical yield (RCY) of 24±1% (non-decay corrected (NDC)) within a synthesis time of 30 min. The [(18)F]SiFB produced by automated synthesis was then used for the (18)F-labeling of rat serum albumin (RSA) as a proof of applicability.

The influenza A(H5N1) epidemic at six and a half years: 500 notified human cases and more to come.

Since November 2003, the epidemic intelligence team at the French Institut de Veille Sanitaire has been gathering data on influenza A(H5N1) circulation in poultry and on human cases worldwide. As Indonesia notifies the world's 500th case to the World Health Organization, we discuss the epidemiological situation and trends of A(H5N1) influenza. Although the overall number of cases reported worldwide has decreased, influenza A(H5N1) continues to circulate intensely in some countries and more cases are to be expected, especially in Egypt and Indonesia.

Coliforms and other microbial indicators occurrence in water and biofilm in full-scale distribution systems.

Biofilm and microbial water quality were studied in four middle size full-scale distribution systems (DS) in France serving 5,000-30,000 inhabitants (maximum residence time 23-160h) through three sampling campaigns over 1 year. Three of these DSs were chosen because of a quite high occurrence of bacterial indicators (i.e. total coliforms), the last DS was considered as a reference. Biofilm was studied on cast iron coupons incubated for more than 1 month in devices continuously fed with water from the DS in conditions imitating those met in DS. The devices were located at different points (4-6) along each DS. The abundance of bacteria in biofilm was estimated by heterotrophic plate counts (HPC) after detachment of the biofilm from the support by sonication. Microbiological water quality was estimated in parallel; analysis of total coliforms, E. coli, enterococci and anaerobic sulphide-reducing bacteria spores (ASRB spores) was carried out in biofilm and water. Over the period of the study, 171 water samples and 57 biofilm samples were collected. Over these 171 waters, 19 (11%) were positive for at least one of the measured indicators while two biofilm samples were positive (3.5%). Significant differences were observed in the levels of contamination between the DSs. High residence time in the DS, low disinfectant residual and high temperature increased the risk of indicator occurrence in the water phase. Due to the low number of biofilm samples positive for bacterial indicators, the data collected in the present study did not allow observation of a direct association between biofilm and water contaminations, even if the occurrence of indicators in water appeared on DSs with the highest density of biofilm (HPC).

Release of organic matter in a discontinuously chlorinated drinking water network.

The effects of discontinuous chlorination on the characteristics of the water in a pilot drinking water distribution network were investigated. The release or consumption of organic matter (as dissolved organic carbon, DOC) following chlorination and non-chlorination periods were estimated, as were changes in bacterial cell production. In each unchlorinated network 0.3 mg DOCl(-1) was consumed and the average cell production was approximately 1.3 x 10(5) cells ml(-1). In discontinously chlorinated networks (chlorine treatment: 3.3 mg Cl2l(-1), chlorine residual: 0.1 mg Cl2l(-1)) the DOC release (DOCout-DOCin) was between 0.1 and 0.2 mg Cl(-1). Biomass production (cells(out)-cells(in)) during this chlorination period was lower (approximately 2 x 10(4) cells ml(-1)). The delay before DOC was released in chlorinated networks appeared to be less than 24 h, which corresponds to one hydraulic residence time. Likewise, when chlorination was stopped, 24 h or less were required before an efficient DOC removal was resumed. When chlorination was prolonged the observed release of DOC was progressively reduced from 0.2 mg l(-1) to zero, thus after 6 weeks of continuous chlorination the DOCin was equivalent to the DOCout.

[Histological and clinical forms of the eosinophilic cellulitis]. / Formes anatomocliniques du syndrome de Wells.

BACKGROUND: Wells' syndrome is characterized by clinical features of cellulitis and a histological picture of eosinophilic infiltrate of the dermis with some "flame" figures. PATIENTS AND METHODS: The clinical and histological features of nine patients with Wells' syndrome seen from 1988 to 1998 were retrospectively reviewed. RESULTS: The clinical features of the nine patients (five men and four women) were urticaria (n=1), cellulitis (n=2), annular plaques (n=3), vesiculo-bullous lesions (n=2) and edema of the face with nodules of the conjunctiva (n=1). Histological examination of skin biopsies showed an eosinophilic infiltrate of the dermis associated with some "flame" figures in all cases. The infiltrate was located in the superficial or deep dermis in accordance with the different clinical features. One patient developed a non Hodgkin lymphoma and presented successively: a Wells' syndrome, a leucocytoklastic vasculitis and a Sweet's syndrome. Numerous treatment were used: topical corticosteroids, H1-antihistamines, dapsone and systemic corticosteroids. Two patients relapsed after treatment withdrawal. DISCUSSION: This study demonstrated a wide polymorphism of the clinical and histological features of Wells' syndrome. The clinical features seem to depend on the location of the dermal infiltrate, suggesting the existence of a spectrum of eosinophilic dermatoses, like in neutrophilic dermatoses. The successive occurrence of vasculitis, Wells' syndrome and Sweet'syndrome in a patient suggests an overlap between these diseases. Systemic corticosteroids are the most effective treatment, but may lead to a corticosteroid dependence.

Chlorine inactivation of Sphingomonas cells attached to goethite particles in drinking water.

Bacteria in drinking water, attached or not attached to goethite particles, were disinfected with chlorine. No additional protection was provided to the bacteria by their attachment to particles, and the limited efficiency of inactivation by chlorine was attributed to the presence of bacterial aggregates in both types of suspension.

Posterior approach with Louis plates for fractures of the thoracolumbar and lumbar spine with and without neurologic deficits.

STUDY DESIGN: This is a retrospective revue of the long-term outcome of posterior Louis plate fixation for the treatment of irreducible or ligamentously unstable fractures of the thoracolumbar and lumbar spine with or without neurologic deficit. OBJECTIVES: To determine the clinical, radiologic, and functional status of patients who underwent posterior fracture fixation with Louis plates and to evaluate the effect of instrumentation length on the construct's ability to maintain reduction of the fracture. SUMMARY OF BACKGROUND DATA: Since the development of pedicle screw fixation described by Roy-Camille, there has been a rapid evolution in the number and complexity of systems available for posterior spinal stabilization. Along with this escalation in implant and instrument sophistication, there has been a corresponding increase in implant cost. To date, no series has been reported of the clinical, radiologic, and functional results of posterior instrumentation with semirigid Louis plates for spinal fractures. METHODS: A retrospective review of spinal fractures from T11 to L5 treated since 1985 by posterior plate fixation showed that 56 patients (37 men and 19 women) with an average age of 34 years and a minimum follow-up of 2 years (average, 41 months) were available for review. There were 36 burst fractures, 4 Chance fractures and 16 fracture-dislocations treated. Forty-three patients had neurologic injury. RESULTS: Before surgery, vertebral kyphosis averaged 15 degrees, improved to 5 degrees with reduction, and reached a steady 10 degrees at final follow-up. Similarly, corrected kyphosis initially averaged 12 degrees but improved to 0.5 degree with reduction, and was 10 degrees at final follow-up. Vertebral canal compromise initially averaged 50% but was reduced to 13% with surgery and 6% at 1 year. There was no significant difference between the ability of short and long constructs to maintain reduction. Eighty-eight percent of patients with neurologic injury improved at least one Frankel grade with treatment. Functional and pain evaluation by the Denis scale showed 25 patients rated P1, 25 rated P2, and 6 rated P3. Twenty-eight were rated W1, 16 were W3, and 12 were W5 at last follow-up. CONCLUSIONS: Fractures of the thoracolumbar spine can be treated effectively with the semirigid Louis plating system. Because of its low cost and ease of insertion, the Louis system is an excellent choice for short arthrodesis and instrumentation of these fractures. Although there is some loss of reduction when compared with more rigid systems, there is no functional compromise in these patients.

Blood clearance kinetics and liver uptake of mononucleosomes in mice.

Nucleosomes generated by apoptosis have become of considerable interest in relation to pathogenesis of systemic lupus erythematosus in mice and humans. Therefore, the fate of circulating mononucleosomes was examined in normal C57Bl/6J mice. The mononucleosomes were prepared from chicken erythrocytes and radiolabeled on the histone component. The removal of nucleosomes from circulation at doses less than 11 micrograms of injected mononucleosomes was rapid, but with increasing doses of injected nucleosomes, the slopes of the removal curves decreased. Liver was the major organ for removal of circulating nucleosomes, accounting for 71.0 to 84.7% of nucleosomes removed from circulation at 10 min. After i.v. injection of nucleosomes, 0.52 +/- 0.15% localized in kidneys. With prior i.v. injection of histones, the glomerular localization of mononucleosomes increased threefold. The clearance of mononucleosomes was decreased sixfold by concurrent injection of ssDNA. These studies show that in mice, circulating mononucleosomes are handled similar to DNA, and they do not avidly localize in glomeruli unless histones have already bound to renal glomeruli.

Bone pain that responds to calcium channel blockers. A retrospective and prospective study of transplant recipients.

A severe episodic bone pain syndrome of unknown cause was first described in renal transplant recipients in France and associated with the use of cyclosporine. We have retrospectively and prospectively evaluated this pain syndrome in our transplant patients. This pain is bilateral, of acute onset and episodic, primarily involving the knees and/or ankles. It usually occurs at night or with recumbency, and is often relieved with elevation or walking. Physical examination of the affected areas is unrevealing. Kidney, liver, pancreas, heart, lung and combined organ transplant recipients on cyclosporine were available from the University of Washington Transplant Services and were retrospectively evaluated by chart review (n = 351) or prospective clinical evaluation (n = 38) for evidence of this clinical syndrome. In the retrospective chart review, 19.1% of patients had episodes of bone pain. The highest prevalence occurred in renal transplant recipients (27.7%). The bone pain syndrome was documented in the charts of 14 patients who subsequently developed significant osteonecrosis. Prospectively, 21 of 22 patients with the bone pain syndrome experienced complete relief of this pain upon treatment with calcium channel blockers (95.4% response rate). The pathophysiology of this bone pain syndrome is unknown, although its response to the vasodilatory effects of calcium channel blockers suggests a vascular etiology.

OmniMed II: a new system for use with the emphasis erodible mask.

Clinical experience has shown the emphasis erodible mask to be an effective method for performing photorefractive keratectomy (PRK) using an eyecup placed in contact with the corneal surface. A new system (OmniMed II) which incorporates the erodible mask as an element in the optical delivery system has been developed for performing photorefractive keratectomy. With this new configuration the eyecup is no longer used. We describe in detail the advantages of the erodible mask, the associated hardware of the optical delivery system, and the mask shape transfer process.

Antigens of varying size persist longer in subepithelial than in subendothelial immune deposits in murine glomeruli.

The kinetics of removal of immune deposits from the subendothelial and subepithelial areas of glomeruli were analyzed in mice. Radiolabeled, cationized Ag of different molecular size, including human serum albumin, bovine thyroglobulin, and human IgM, were used to form the immune deposits in mouse glomeruli with specific, purified rabbit antibodies to these proteins. The disappearance curves of the radiolabeled Ag from glomeruli consisted of two exponential components. The immune deposits and their location in glomeruli were identified by immunofluorescence and electron microscopy. The t1/2 of disappearance of immune deposits were assigned to subendothelial or subepithelial deposits on the basis of ultrastructural observations. The t1/2 of subendothelial immune deposits ranged from 0.6 to 1.9 days with the three different Ag-antibody systems. In contrast, the t1/2 of the subepithelial immune deposits ranged from 9.32 to 231 days. The cationized human serum albumin in subepithelial areas had the longest t1/2, and this was not altered by the endogenous immune response to the injected materials, as determined in studies with nude mice. The results constitute formal documentation of the prolonged t1/2 of an exogenous Ag in glomerular immune deposits. The described approach can serve to examine variables that alter this prolonged presence of subepithelial immune deposits in glomeruli.

Mapping of activation, recovery and activation-recovery intervals in the human heart.

Peroperative epicardial mapping of activation, recovery and activation-recovery intervals in the human heart has been performed in a group of 12 patients. These patients had the coronary disease but electrocardiograms with normal characteristics. For this mapping, 240 unipolar electrograms were simultaneously recorded with the system SATAPEC built in our laboratory. The results confirm the classical data obtained on the dog heart. In particular, it was well established that the duration of activation corresponding to activation-recovery intervals is shorter at the base than at the apex of the posterior surface of the heart. With SATAPEC it is very easy to obtain epicardial mapping of electrical activity in a few minutes during open heart surgery.

Autoantibodies to the collagen-like region of C1Q deposit in glomeruli via C1Q in immune deposits.

The autoantibodies to the collagen-like region of C1q (CLR), purified from two patients with systemic lupus erythematosus, deposited in mouse glomeruli when human C1q was present in antigen-antibody complexes in glomeruli. The immune deposits with C1q in mouse glomeruli were achieved by the administration of cationized immune complexes containing human C1q. The presented data suggest that the autoantibodies to CLR could enhance the pathogenic role of immune complexes deposited in glomeruli by binding to C1q in immune deposits. These findings may explain the association of autoantibodies to CLR with proliferative lupus nephritis.

Assessment with potential mapping of the cardiac protective effect of a drug. Example of trimetazidine.

SATAPEC is a computerized system built in the Laboratory for recording over 256 multiplexed channels and for automatic processing of cardiac electrical potentials. The results are printed out in the form of maps (either in black and white or in color) showing potential distributions, depolarization, repolarization, duration of activation, or various other analog-digital data. The cardioprotective ability of a drug may be assessed with SATAPEC. As an example, the effect of trimetazidine (TMZ) is examined using 2 groups of 12 rabbits (one group serving as a control and the second group pretreated with TMZ). An elastomer mesh with 240 regularly spaced chlorided silver wire electrode is placed around the ventricles following thoracotomy. A ligature is made starting from the anterior interventricular artery (AIV). Recordings are taken from the 240 unipolar epicardial electrograms (reference potential taken at Wilson terminal) 1 min before ligation and then every min for 8 min following ligation. Once the electrograms are plotted and any aberrant tracings eliminated, ST variation is calculated automatically. Files containing ST variations at different instants are stored in the computer memory and the mean sigma ST/240 curves of the two groups of rabbits are plotted versus time. Mean electric potential maps, obtained by aligning all of the individual maps, are then printed. With these maps the location and extent of the epicardial injured area can be visualized. Pretreatment with TMZ (2.5 mg/kg) has shown a beneficial effect on ischemia injury.