Effect of proteases against biofilms of Staphylococcus aureus and Staphylococcus epidermidis.

UNLABELLED: Biofilms play a key role in bacterial resistance against antibacterial agents-an issue that causes multiple problems in medical fields, particularly with Staphylococcus biofilms that colonize medical indwelling devices. The literature reports several anti-biofilm strategies that have been applied in medicine. Disrupting the biofilm formation process creates new sites open to colonization by treatment-generated planktonic bacteria, so efforts have turned to focus on strategies to prevent and control the initial Staphylococci adhesion. Here, we investigated the preventive activities of three commercial proteases (Flavourzyme, Neutrase and Alcalase) against biofilm formation by two Staphylococcus strains. Some proteolytic extracts revealed interesting results with Staphylococcus epidermidis and Staphylococcus aureus aureus biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: Three proteases were tested against Staphylococcus aureus and Staphylococcus epidermidis biofilms in standard conditions. The Flavourzyme containing a mix of Aspergillus orizae endo- and exoproteases demonstrated significant efficacy against Staph. epidermidis biofilm formation. These results could prove valuable in the effort to develop simple anti-biofilm methods.

[Severe cytomegalovirus-associated protein losing gastropathy in an immunocompetent adult]. / Entéropathie exsudative sévère compliquant une primo-infection à cytomégalovirus chez un adulte immunocompétent.

INTRODUCTION: Clinical expression of cytomegalovirus (CMV) infection is numerous and depends on the immune status of the host. In immunocompromised patients, CMV disease corresponds to reactivation of the virus with tissue damage responsible for significant morbidity and mortality. In immunocompetent adults, primary CMV infection is usually asymptomatic but may rarely result in severe organ damage. OBSERVATION: We report the case of a 28-year-old man, considered immunocompetent and without medical history, presenting with a primary CMV infection revealed by a severe protein-losing gastroenteropathy (serum albumin level: 18.9g/L). The CT scan showed a gastro-duodeno-jejuno-ileitis pattern. Upper intestinal endoscopy revealed diffuse ulcerated and congestive gastritis predominantly in the fundus. Histological analysis of the biopsies showed no viral inclusion and no immunohistochemical staining reaction, but a high viral load (5.61 log). Treatment was symptomatic associated with anticoagulation due to the major hypoalbuminemia. The patient slowly recovered, and normal clinical examination and blood tests were observed two months after the onset of symptoms. CONCLUSION: Severe CMV organ involvement in the immunocompetent patient is very rare but potentially serious. Digestive involvement complicated by exudative enteropathy is possible. Evolution is usually favorable without the need for antiviral treatment.

Plants arabinogalactans: From structures to physico-chemical and biological properties.

Arabinogalactans (AGs) are plant heteropolysaccharides with complex structures occasionally attached to proteins (AGPs). AGs in cell matrix of different parts of plant are freely available or chemically bound to pectin rhamnogalactan. Type I with predominantly ß-d-(1 â†’ 4)-galactan and type II with ß-d-(1 â†’ 3) and/or (1 â†’ 6)-galactan structural backbones construct the two main groups of AGs. In the current review, the chemical structure of AGs is firstly discussed focusing on non-traditional plant sources and not including well known industrial gums. After that, processes for their extraction and purification are considered and finally their techno-functional and biological properties are highlighted. The role of AG structure and function on health advantages such as anti-tumor, antioxidant, anti-ulcer- anti-diabetic and other activites and also the immunomodulatory effects on in-vivo model systems are overviewed.

RGK GTPase-dependent CaV2.1 Ca2+ channel inhibition is independent of CaVbeta-subunit-induced current potentiation.

RGK (Rad-Gem-Rem) GTPases have been described as potent negative regulators of the Ca(2+) influx via high-threshold voltage-activated Ca(2+) channels. Recent work, mostly performed on Ca(V)1.2 Ca(2+) channels, has highlighted the crucial role played by the channel auxiliary Ca(V)beta subunits and identified several GTPase and beta-subunit protein domains involved in this regulation. We now extend these conclusions by producing the first complete characterization of the effects of Gem, Rem, and Rem2 on the neuronal Ca(V)2.1 Ca(2+) channels expressed with Ca(V)beta(1) or Ca(V)beta(2) subunits. Current inhibition is limited to a decrease in amplitude with no modification in the voltage dependence or kinetics of the current. We demonstrate that this inhibition can occur for Ca(V)beta constructs with impaired capacity to induce current potentiation, but that it is lost for Ca(V)beta constructs deleted for their beta-interaction domain. The RGK C-terminal last approximately 80 amino acids are sufficient to allow potent current inhibition and in vivo beta-subunit/Gem interaction. Interestingly, although Gem and Gem carboxy-terminus induce a completely different pattern of beta-subunit cellular localization, they both potently inhibit Ca(V)2.1 channels. These data therefore set the status of neuronal Ca(V)2.1 Ca(2+) channel inhibition by RGK GTPases, emphasizing the role of short amino acid sequences of both proteins in beta-subunit binding and channel inhibition and revealing a new mechanism for channel inhibition.

Purification and Valorization of Waste Cotton Seed Oil as an Alternative Feedstock for Biodiesel Production.

This article is focused on the production of biodiesel from the waste cotton seed oil (WCSO), after purification, as an alternative to fossil fuels. Waste oil was collected from Sodecoton, a factory producing cotton seed oil in the Far North Cameroon. The WCSO was subjected to purification using activated coal, followed by transesterification under basic conditions (potassium hydroxide (KOH)), using methanol and ethanol. Some physico-chemical properties of biodiesel, such as absorbance of waste and purified oil, density, viscosity, water content, acid value, and its energy content were determined. The result of treating the WCSO with activated coal indicated that purification efficiency of activated coal increased with the contact time and the mass of the absorbent. Absorbance results directly proved that activated coal removed unwanted components. In the same way, activated coal concentration and exposure time influenced the level of free fatty acids of WCSO. The yield of methyl ester was 97%, while that of ethyl ester was 98%. The specific gravity at 25 °C was 0.945 ± 0.0601. An evaluation of the lower calorific value (PCI) was done in order to study the energy content of biodiesel. This was found to be a value of 37.02 ± 3.05 MJ/kg for methyl ester and 36.92 ± 7.20 MJ/kg for ethyl ester. WCSO constitutes feedstock for high volume, good quality, and sustainable production of biodiesel, as well as a realistic means of eliminating the pollution resulting from the indiscriminate disposal of waste oils from both household and industrial users.

[Respiratory consequences of inhalation of adulterated cannabis]. / Conséquences respiratoires de l'inhalation d'herbe de cannabis frelatée.

Cannabis is widely smoked in Europe and its increasing use is becoming a major public health problem. Adulterating cannabis with glass beads or sand is a new trick used by dealers to increase the weight and boost profits. These recent practices are not without danger. We report two cases of respiratory symptoms related to the use of this kind of adulterated cannabis. The first case is a 33 year-old patient admitted for an acute inhalation pneumonitis secondary to smoking cannabis adulterated with grit sand. The CT scan showed patchy ground-glass opacities, mainly in the upper lobes. A broncho-alveolar lavage, examined under polarized light, revealed birefringent intracellular particles, identified as silica, in alveolar macrophages. Spontaneously clinical and radiological improvements were observed after stopping the use of contaminated cannabis. The second patient, who smoked cannabis mixed with glass beads, described epistaxis, mouth ulcers, sore throats and cough. CT scan and BAL were normal. Adulteration of cannabis with microscopic glass beads, alone or mixed with sand, is a recent and widespread practice in Europe. These anecdotal reports prompted the French Department of Health to advise cannabis smokers of the harmfulness of these contaminants.

Fluorescence in situ hybridization characterization of ider(20q) in myelodysplastic syndrome.

Isochromosome of the long arm of chromosome 20 with loss of interstitial material [ider(20q)] is a variant of deletion of chromosome 20q and a rare abnormality in myelodysplastic syndrome (MDS). We studied seven cases with an ider(20q) in MDS. Fluorescence in situ hybridization (FISH) studies showed all proximal breakpoints to be consistently located in 20q11.21 band whereas distal breakpoints were variable. Amplification of HCK, TNFRSF6B and DIDO1 genes included in retained regions associated with loss of tumour suppressor genes in deleted regions could explain cell tumour progression and possibly the less favourable prognosis of ider(20q) compared with del(20q).

Organic synthesis in soft wall-free microreactors: real-time monitoring of fluorogenic reactions.

A new approach of "laboratory on a chip" (LOC) devoted to organic synthesis based on electrically operated ionic liquid microdroplets used as air-stable "soft" microreactors was recently introduced. A number of challenging issues have yet to be addressed to allow standard macroscale organic chemistry to be directly transposed in these nonclassical microreactors. In particular, since standard (i.e., magnetical or mechanical) stirring methods are prohibited in such wall-free microreactors, effective alternatives have to be developed to circumvent mass-transfer limitations. With this aim in mind, a fluorogenic version of a click chemistry reaction was developed to evaluate the efficiency of alternative mixing methods on the reaction kinetics. We demonstrate that the combination of chaotic advection created by surface acoustic waves combined with a temperature increase (Marangoni effect) leads to the same kinetics regime as in standard macroscale conditions. This opens the route for application of the new generation of LOC to efficient organic synthesis in microscale.

New monolithic enzymatic micro-reactor for the fast production and purification of oligogalacturonides.

Fast production and purification of alpha-(1,4)-oligogalacturonides was investigated using a new enzymatic reactor composed of a monolithic matrix. Pectin lyase from Aspergillus japonicus (Sigma) was immobilized on CIM-disk epoxy monolith. Studies were performed on free pectin lyase and immobilized pectin lyase to compare the optimum temperature, optimum pH, and thermal stability. It was determined that optimum temperature for free pectin lyase and immobilized pectin lyase on monolithic support is 30 degrees C, and optimum pH is 5. Monolithic CIM-disk chromatography is one of the fastest liquid chromatographic method used for separation and purification of biomolecules due to high mass transfer rate. In this context, online one step production and purification of oligogalacturonides was investigated associating CIM-disk pectin lyase and CIM-disk DEAE. This efficient enzymatic bioreactor production of uronic oligosaccharides from polygalacturonic acid (PGA) constitutes an original fast process to generate bioactive oligouronides.

Purification of oligouronides by immobilized L-histidine pseudoaffinity chromatography.

Selective purification of alpha-(1,4)-oligogalacturonides was investigated using several pseudobioaffinity chromatography matrix with aminoacid L-histidine as pseudobiospecific ligand: (1) sepharose 4B-bisoxyran-histidine, (2) sepharose 4B-epoxy-histidine, (3) silica-oxyran-histidine and (4) CIM-disk-EDA-histidine. These anionic oligosaccharides prepared by enzymatic and chemical cleavage of polygalacturonic acid were used as models sugar in order to optimize the adsorption and elution parameters for a selective purification of bioactive oligouronides. Monolithic CIM-disk chromatography is one of the fastest liquid chromatographic method using for separation and purification of biomolecules thanks to high mass transfer rate. In this way, this new monolithic CIM-disk system with L-histidine immobilized: immobilized histidine affinity chromatography (IHAC) constitutes a good tool allowing the fast and selective purification of bioactive oligouronides.

Production of oligoglucuronans using a monolithic enzymatic microreactor.

A glucuronan lyase (EC 4.2.2.14) was immobilized on a monolithic Convective Interaction Media (CIM) disk. The immobilization yield was equal to 29% of the initial activity and 35% of the initial protein amount. Degradations of three glucuronans with various O-acetylation degrees were investigated and compared with degradations using free enzyme. The immobilized glucuronan lyase was inhibited by the O-acetylation degree like the free enzyme. (1)H NMR analyses were used to study the O-acetylation degree of oligoglucuronans and demonstrated that the average degrees of polymerization were inclusive between 4 and 13 after 24h of degradation. This first immobilization of a glucuronan lyase constitutes a new tool to produce oligoglucuronans.

Oligogalacturonans production by free radical depolymerization of polygalacturonan.

The large production of acidic oligosaccharides was investigated by non-enzymatic depolymerization of polygalacturonic acid (PGA) using free hydroxyl radical hydrolysis process from H2O2/copper (II) system. A large amount of oligogalacturonides (OGAs) with degrees of polymerization up to 6 were fractionated, and characterized by ESI-Q/TOF-MASS spectrometry and 1H NMR spectroscopy. This efficient production of uronic oligosaccharides from PGA constitutes an original process to produce bioactive compounds in large scale up.

Thermal decomposition of expanded polystyrene in a pebble bed reactor to get higher liquid fraction yield at low temperatures.

Expanded polystyrene is one of the polymers produced in large quantities due to its versatile application in different fields. This polymer is one of the most intractable components in municipal solid waste. Disposal of polymeric material by pyrolysis or catalytic cracking yields valuable hydrocarbon fuels or monomers. Literature reports different types of reactors and arrangements that have uniform temperatures during pyrolysis and catalytic cracking. The present study focuses on reducing the temperature to maximize the quantity of styrene monomer in the liquid product. A bench scale reactor has been developed to recover the styrene monomer and other valuable chemicals. Experiments were carried under partial oxidation and vacuum conditions in the temperature range of 300-500 degrees C. In the pyrolysis optimization studies, the best atmospheric condition was determined to be vacuum, the pyrolysis temperature should be 500 degrees C, yield of liquid product obtained was 91.7% and yield of styrene obtained was 85.5%. In the characterization studies, distillation and IR spectroscopy experiments were carried out. The remaining of the liquid product comprises of benzene, ethyl benzene, and styrene dimers and trimers.

[Developping real time lymph node aspiration under endobronchial ultrasound control. One year's French experience in two different pulmonary departments]. / Mise en place de l'échoendoscopie bronchique avec ponction ganglionnaire en pneumologie: expérience française de deux centres.

INTRODUCTION: The histology and staging of bronchial carcinoma determines the treatment options for the condition. Endobronchial ultrasound allows the needle aspiration of mediastinal lymph nodes or pulmonary neoplasia where there is tracheo-bronchial contact under visual control. This procedure is aid for diagnosis and for mediastinal staging. French pulmonary departments have been slow to introduce this technique compared to other countries. METHODS: All Endobronchial ultrasound procedures performed during 2007 were retrospectively analysed in two pulmonary centres. The indications, practice management, complications, and diagnostic yield were reported. RESULTS: 103 Endobronchial ultrasound procedures were performed, in the majority under local anaesthesia in out-patients. Real time needle aspiration was performed only in 92 patients. Only 11 procedures were performed for mediastinal staging prior to thoracic surgery. 12.6% of patients had minor complications. 136 lymph node stations were sampled in 92 patients, but only 97 (70.3%) in 63 patients were judged to be 'satisfactory"(malignant cells and/or lymphocytes on cytology results). CONCLUSIONS: It is difficult to rapidly reach the diagnostic yield reported in literature. We think that appropriate training in the technique is of great importance.

[Intravascular lymphoma revealed by generalized arborescent telangiectasia and repeated venous thrombosis]. / Thromboses veineuses récidivantes et télangiectasies arborescentes généralisées révélatrices d'un lymphome intravasculaire.

BACKGROUND: Intravascular lymphoma is a rare form of non-Hodgkin's lymphoma characterized by proliferation of lymphoid cells within the lumen of small veins, small arteries and capillaries. CASE REPORT: A 79-year-old man presented with repeated superficial venous thrombosis of the lower limbs associated with diffuse telangiectasia of the trunk, upper arms and thighs but with normal epidermis. Screening for thrombophilia and neoplasm were negative. The patient subsequently developed abdominal pain, lower-limb oedema, deterioration in performance status and rapidly increasing telangiectasia with the appearance of generalized oedematous cutaneous induration. Increased LDH and anaemia were observed without other blood count anomalies. Intravascular lymphoma was diagnosed on a skin biopsy with telangiectasia and oedema. After eight courses of treatment with rituximab-CHOP the outcome was good. DISCUSSION: A few telangiectasias associated with nodules or infiltrated plaques are often a clinical manifestation of intravascular lymphoma. The dermatological presentation described here is interesting for two reasons: on the one hand, telangiectasias were initially isolated in normal epidermis without any infiltration for more than one year and, on the other hand, the skin infiltration seen subsequently was very extensive and marked. Moreover, although microthrombi are frequent in small and medium-sized blood vessels, thrombosis of large vessels is rarely described in intravascular lymphoma.

Hypereosinophilia and epigastralgia after visiting Tunisia: a rare differential diagnosis of helminthiases.

Causes of hypereosinophilia among travelers returning from North Africa are dominated by helminth infections, especially when associated with gastrointestinal signs. Non-infectious causes must nonetheless be investigated after negative microbiological assessment and failure of a broad empiric antiparasite treatment. We report the case of a young man with epigastralgia and major weight loss since a stay in Tunisia. Empiric treatment with albendazole was not successful. Eosinophilic gastroenteritis was diagnosed and resolved under corticosteroid treatment.

Hippocampal Deformations and Entorhinal Cortex Atrophy as an Anatomical Signature of Long-Term Cognitive Impairment: from the MCAO Rat Model to the Stroke Patient.

Stroke patients have an elevated risk of developing long-term cognitive disorders or dementia. The latter is often associated with atrophy of the medial temporal lobe. However, it is not clear whether hippocampal and entorhinal cortex atrophy is the sole predictor of long-term post-stroke dementia. We hypothesized that hippocampal deformation (rather than atrophy) is a predictive marker of long-term post-stroke dementia on a rat model and tested this hypothesis in a prospective cohort of stroke patients.Male Wistar rats were subjected to transient middle cerebral artery occlusion and assessed 6 months later. Ninety initially dementia-free patients having suffered a first-ever ischemic stroke were prospectively included in a clinical study. In the rat model, significant impairments in hippocampus-dependent memories were observed. MRI studies did not reveal significant atrophy of the hippocampus volume, but significant deformations were indeed observed-particularly on the ipsilateral side. There, the neuronal surface area was significantly lower in ischemic rats and was associated with a lower tissue density and a markedly thinner entorhinal cortex. At 6 months post-stroke, 49 of the 90 patients displayed cognitive impairment (males 55.10%). Shape analysis revealed marked deformations of their left hippocampus, a significantly lower entorhinal cortex surface area, and a wider rhinal sulcus but no hippocampal atrophy. Hence, hippocampal deformations and entorhinal cortex atrophy were associated with long-term impaired cognitive abilities in a stroke rat model and in stroke patients. When combined with existing biomarkers, these markers might constitute sensitive new tools for the early prediction of post-stroke dementia.

Antioxidant Activities of Peptoid-Grafted Chitosan Films.

The aim of this study was to investigate the possibility of immobilizing peptoid on chitosan film in order to generate new active material. Chitosan films have been grafted for the first time with short-length peptoid oligomers displaying antioxidant activities. The antioxidant activity of the selected peptoids was initially investigated with the DPPH assay and hydroxyl radical procedure. The metal chelating capacity of peptoids was also evaluated prior to their covalent attachment to chitosan. The benefit of chitosan functionalization with respect to its intrinsic antioxidant properties was finally evaluated in the present study. Interestingly, an increase of up to 90 % of the antioxidant activity of chitosan was observed.

Structural characterization and rheological properties of a galactomannan from Astragalus gombo Bunge seeds harvested in Algerian Sahara.

A water soluble polysaccharide (WSP) was extracted and purified from Astragalus gombo seeds (Fabaceae) harvested in Septentrional Sahara (Ouargla, Algeria) with a yield of 6.8% (w/w of the dry seed ground). It was characterized by gas chromatography coupled to the mass spectrometry (GC-MS), size exclusion chromatography with Multi-Angle Light Scattering analysis (SEC-MALLS), high-resolution 1H and 13C NMR, and rheological measurements. The structural characterization indicated that this WSP fraction is a galactomannan with a mannose/galactose ratio of 1.7 formed by a backbone of ß-(1,4)-d-mannopyranosyl residues (63%) substituted at O-6 position by a single α-galactopyranose residue (37%). SEC-MALLS analysis revealed that this galactomannan has an average molecular mass (Mw) of 1.1×106g/mol, an intrinsic viscosity of 860mL/g and, a random coil conformation structure. Rheological analysis in semi diluted regimes shown pseudo-plastic and viscoelastic behaviour.

Identification of genes overexpressed in tumors through preferential expression screening in trophoblasts.

Early trophoblastic cells share several features with neoplastic cells. Based on that observation, we attempted to identify genes overexpressed in tumors by analyzing genes preferentially expressed in trophoblasts. A subtracted library enriched in complementary DNA from early cytotrophoblasts was constructed, and the expression level of selected recombinants was analyzed on a large panel of normal and tumor tissues. The library was prepared using a polymerase chain reaction-based complementary DNA subtraction method with 6-week amenorrhea cytotrophoblast endoplasmic reticulum-bound RNA as target, and a mixture of complementary DNA prepared from terminal placenta and activated T-lymphocytes as driver. Two rounds of screening were performed to isolate clones preferentially expressed in early placenta. From a total number of recombinant clones estimated at 32,000 in the subtracted library, 594 inserts were analyzed by Southern blot and 21 sequences were isolated as corresponding to genes highly expressed in early placenta. Eleven encoded known molecules, such as carcinoembryonic antigen, human chorionic gonadotropin, and mitochondrial rRNAs. Ten sequences represented novel genes. Northern blot analysis confirmed that most of these genes were preferentially expressed in early trophoblast in comparison to terminal placenta. Three clones that gave detectable hybridization signals on total RNA were extensively studied and were found to be overexpressed in various tumors. Two of these clones, designated B9 and E4, were later identified as corresponding to genes coding for the putative ribosomal protein S18 and the bifunctional enzyme ADE2H1 involved in purine biosynthesis, respectively. Expression of the third clone, E9, was increased up to 10-fold in breast cancer tissues in comparison with normal counterparts. Present results confirm that many genes expressed in the trophoblast are overexpressed in malignant cells. This approach could provide a general targeted method for the identification of genes overexpressed in various neoplastic cell types.