Candidiasis, caused mainly by Candida albicans, a natural commensal of the human digestive tract and vagina, is the most common opportunistic fungal infection at the mucosal and systemic levels. Its high morbi–mortality rates have led to considerable research to identify the molecular mechanisms associated with the switch to pathogenic development and to diagnose this process as accurately as possible. Since the 1980s, the advent of monoclonal antibody (mAb) technology has led to significant progress in both interrelated fields. This linear review, intended to be didactic, was prompted by considering how, over several decades, a single mAb designated 5B2 contributed to the elucidation of the molecular mechanisms of pathogenesis based on β-1,2-linked oligomannoside expression in Candida species. These contributions starting from the structural identification of the minimal epitope as a di-mannoside from the β-1,2 series consisted then in the demonstration that it was shared by a large number of cell wall proteins differently anchored in the cell wall and the discovery of a cell wall glycoplipid shed by the yeast in contact of host cells, the phospholipomannan. Cytological analysis revealed an overall highly complex epitope expression at the cell surface concerning all growth phases and a patchy distribution resulting from the merging of cytoplasmic vesicles to plasmalema and further secretion through cell wall channels. On the host side, the mAb 5B2 led to identification of Galectin-3 as the human receptor dedicated to β-mannosides and signal transduction pathways leading to cytokine secretion directing host immune responses. Clinical applications concerned in vivo imaging of Candida infectious foci, direct examination of clinical samples and detection of circulating serum antigens that complement the Platelia Ag test for an increased sensitivity of diagnosis. Finally, the most interesting character of mAb 5B2 is probably its ability to reveal C. albicans pathogenic behaviour in reacting specifically with vaginal secretions from women infected versus colonized by this species as well as to display higher reactivity with strains isolated in pathogenic circumstances or even linked to an unfavourable prognosis for systemic candidiasis. Together with a detailed referenced description of these studies, the review provides a complementary reading frame by listing the wide range of technologies involving mAb 5B2 over time, evidencing a practical robustness and versatility unique so far in the Candida field. Finally, the basic and clinical perspectives opened up by these studies are briefly discussed with regard to prospects for future applications of mAb 5B2 in current research challenges.

Ultra-fast cooling for mild therapeutic hypothermia (MTH) has several potential applications, including prevention of post-cardiac arrest syndrome. Ultra-fast MTH by total liquid ventilation (TLV) entails the sudden filling of the lungs with a cold perfluorocarbon liquid and its subsequent use to perform TLV. The present physiological study was aimed at assessing whether pulmonary and systemic hemodynamics as well as lung mechanics are significantly altered during this procedure. Pulmonary and systemic arterial pressures, cardiac output as well as airway resistance and respiratory system compliance were measured during ultra-fast MTH by TLV followed by rewarming and normothermia in six healthy juvenile lambs. Results show that none of the studied variables were altered upon varying the perfluorocarbon temperature from 12 to 41 °C. It is concluded that ultra-fast MTH by TLV does not have any deleterious effect on hemodynamics or lung mechanics in healthy juvenile lambs.

Dermatophytes are an important cause of superficial fungal infection. Direct examination of skin, nail, or hair samples remains essential in diagnosis, as it provides a quick response to the clinician. However, mycological analysis, including direct examination and culture, often lacks sensitivity. The use of stains or fluorochromes may enhance the performance of direct examination. We analyzed 102 samples from patients with suspected dermatophytosis in 4 different diagnostic mycology laboratories. Two reagents, MycetColor® and MycetFluo®, which use Congo red and calcofluor dye, respectively, were evaluated for the direct microscopic examination of skin, hair, and nail specimens. The results were compared to those of culture and conventional direct examination. Both reagents were able to clarify the specimens and also to specifically stain fungal elements. Microscopic examination of the specimens was greatly facilitated with MycetFluo®, which allowed a higher number of positive cases to be detected compared to the other methods.

Total liquid ventilation (TLV) is an emerging mechanical ventilation technique. In this technique, the lungs are filled with liquid perfluorocarbons (PFC) and a liquid ventilator assures ventilation by periodically renewing a volume of oxygenated, CO2 freed and temperature controlled PFC. A huge difference between conventional mechanical ventilation and TLV relates to the fact that PFCs are about 1500 times denser than air. Thus, the PFCs filled lungs turn into an efficient heat exchanger with the circulating blood. One of the most appealing utilization of the lungs as a heat exchanger in TLV is for ultrafast induction of mild therapeutic hypothermia (MTH) for neuroprotection and cardioprotection after ischemia-reperfusion injuries. This study aimed to perform ultrafast MTH induction by TLV in animals up to 25 kg, then perform a fast post-hypothermic rewarming while maintaining proper ventilation. A thermal model of the lamb and liquid ventilator was developed to predict the dynamic and the control strategy to adopt for MTH induction. Two juvenile lambs were instrumented with temperature sensors in the femoral artery, pulmonary artery, oesophagus, right eardrum and rectum. After stabilization in conventional mechanical ventilation, TLV was initiated with ultrafast MTH induction, followed by posthypothermic rewarming. Preliminary results in the two juvenile lambs reveal that the liquid ventilator Inolivent-6.0 can induce MTH by TLV in less than 2.5 min for systemic arterial blood and in less than 10 min for venous return, esophagus and eardrum. Rectal temperature reached MTH in respectively 19.4 and 17.0 min for both lambs. Experimental results were consistent with the model predictions. Moreover, blood gas analysis exhibited that the gas exchange in the lungs was maintained adequately for the entire experiments.

Background: Total liquid ventilation (TLV) consists in filling the lungs with a perfluorocarbon (PFC) and using a liquid ventilator to ensure a tidal volume of oxygenated, CO 2 -free and temperature-controlled PFC. Having a much higher thermal capacity than air, liquid PFCs assume that the filled lungs become an efficient heat exchanger with pulmonary circulation. Objective: The objective of the present study was the development and validation of a parametric lumped thermal model of a subject in TLV. Methods: The lungs were modeled as one compartment in which the control volume varied as a function of the tidal volume. The heat transfer in the body was modeled as seven parallel compartments representing organs and tissues. The thermal model of the lungs and body was validated with two groups of lambs of different ages and weights (newborn and juvenile) undergoing an ultrafast mild therapeutic hypothermia induction by TLV. Results: The model error on all animals yielded a small mean error of -0.1 ± 0.4 °C for the femoral artery and 0.0 ± 0.1 °C for the pulmonary artery. Conclusion: The resulting experimental validation attests that the model provided an accurate estimation of the systemic arterial temperature and the venous return temperature. Significance: This comprehensive thermal model of the lungs and body has the advantage of closely modeling the rapid thermal dynamics in TLV. The model can explain how the time to achieve mild hypothermia between newborn and juvenile lambs remained similar despite of highly different physiological and ventilatory parameters. The strength of the model is its strong relationship with the physiological parameters of the subjects, which suggests its suitability for projection to humans.

Total liquid ventilation (TLV) is an experimental mechanical ventilation technique where the lungs are completely filled with a perfluorocarbon liquid (PFC). It can be used to implement moderate therapeutic hypothermia (MTH) and treat severe respiratory problems. During TLV, the airway pressure must be monitored adequately to avoid overpressure and airway collapses. On the thermodynamic level, rectal, esophageal or tympanic temperature measurements are not suitable (long time constant) to avoid lowering the heart below 30°C. The objective was to design a Y connector positioned at the mouth which integrates the virtual sensors, used by controllers. The first estimates the airway pressure and the second provides the core body temperature. Pressure and RTD sensors were installed in the connector to implement the virtual measurements. In-vitro experiments were done to validate the virtual sensors. In-vivo experiments (on newborn lambs) confirm the accuracy of the airway pressure estimation and of the systemic arterial temperature.

Total liquid ventilation (TLV) is an emerging and promising mechanical ventilation method in which the lungs are filled with a breathable liquid. Perfluorocarbon (PFC) is the predominant liquid of choice due to its high O-2 and CO2 solubility. In TLV, a dedicated liquid ventilator ensures gas exchange by renewing a tidal volume of PFC, which is temperature-controlled, oxygenated and free of CO2. A fundamental difference between TLV and conventional mechanical ventilation relates to the fact that PFCs are approximately 1500 times denser than air. This high density provides PFCs with a large heat capacity, turning the lungs into an efficient heat exchanger with circulating blood. The originality of this study is the development of a lumped thermal model of the body as a heat exchanger coupled to a liquid ventilator. The model was validated with an animal experimentation on a newborn lamb with the Inolivent-5.0 liquid ventilator prototype. TLV was initiated with a fast hypothermia induction, followed successively by a slow posthypothermic rewarming, a fast rewarming and finally a second fast hypothermia induction. Results demonstrate that the model was able to aptly predict, in every phase, the temperature of the lungs, the eardrum, the rectum as well as the various compartments of the liquid ventilator.

Introduction L’administration intrapulmonaire de perfluorocarbones par ventilation liquide totale (VLT) permet un refroidissement ultra-rapide tout en maintenant les échanges gazeux. La VLT hypothermisante (VLTH) a montré des effets bénéfiques neurologiques et cardiaques dans des modèles animaux de choc et d’arrêt cardiaque. L’objectif de ce travail était d’étudier son intérêt sur l’atteinte motrice des membres inférieurs dans un modèle d’ischémie-reperfusion médullaire par clampage aortique. Nous avons évalué l’effet de la profondeur de l’hypothermie (33 vs 36°C) et de la sévérité de l’ischémie (15 vs 20minutes). Matériel et méthodes Des lapins ont été anesthésiés, instrumentés et soumis à une ischémie médullaire par clampage de l’aorte abdominale infra-rénale pendant 15 ou 20min. Ils ont ensuite aléatoirement subi une procédure « témoin » (ventilation conventionnelle et normothermie à 38,5°C) ou une VLTH avec des températures cibles de 33°C ou 36°C. La VLTH était débutée immédiatement après la réperfusion (déclampage). Après 30min de VLTH, l’hypothermie était maintenue durant 120min avant de réchauffer les animaux (1°C/h) et de les réveiller. L’évaluation neurologique était conduite 48h après l’ischémie médullaire à l’aide du score de Tarlov (0 : paraplégie totale ; 5 : fonction motrice normale). Résultats Trente animaux ont été inclus dans cette étude (n=5 par groupe). En conditions témoin, tous les animaux présentaient une paraplégie totale avec un score de Tarlov égal à 0, quelle que soit la durée de l’ischémie initiale (15 ou 20min). Dans les groupes VLTH, la température cible était atteinte très rapidement, c’est-à-dire en moins 15-20min dans tout l’organisme après la réperfusion. Après 15min d’ischémie, la VLTH apportait une amélioration significative de l’atteinte motrice par rapport au groupe témoin, quelle que soit la température étudiée (Fig. 1 : *p<0,05 par rapport au groupe témoin correspondant). Après 20min d’ischémie, une tendance à l’amélioration de la fonction motrice était observée uniquement après une VLTH à 33°C. Discussion En cas d’ischémie-réperfusion médullaire de gravité modérée, l’hypothermie ultra-rapide induite par VLT permet d’atténuer la souffrance neurologique quelle que soit la température cible. A contrario, une température plus basse semble nécessaire pour obtenir un bénéfice en cas d’ischémie-réperfusion médullaire plus sévère. L’hypothermie par VLT pourrait ouvrir des perspectives de protection en cas d’atteinte médullaire post-opératoire lors des chirurgies aortiques majeures.

Les tests de diagnostic rapide (TDR) connaissent un essor dans le diagnostic des maladies infectieuses et parasitaires. En mycologie médicale, ils concernent essentiellement (i) les candidoses avec l’agglutination indirecte de particules de latex sensibilisées avec des anticorps monoclonaux pour l’identification des colonies de Candida, (ii) les cryptococcoses dont le diagnostic par recherche d’antigène capsulaire, présent à forte concentration dans les liquides biologiques, se prête bien à la technique d’immunochromatographie sur membrane (ICM) et (iii) les aspergilloses invasives avec le développement d’ICM utilisant des anticorps monoclonaux différents de ceux reconnaissant les galactomannanes pour éviter les problèmes de réactions faussement positives observées avec les tests ELISA actuels.

Objectives: Total liquid ventilation provides ultrafast and potently neuro- and cardioprotective cooling after shockable cardiac arrest and myocardial infarction in animals. Our goal was to decipher the effect of hypothermic total liquid ventilation on the systemic and cerebral response to asphyxial cardiac arrest using an original pressure- and volume-controlled ventilation strategy in rabbits.Design: Randomized animal study.Setting: Academic research laboratory.Subjects: New Zealand Rabbits.Interventions: Thirty-six rabbits were submitted to 13 minutes of asphyxia, leading to cardiac arrest. After resumption of spontaneous circulation, they underwent either normothermic life support (control group, n = 12) or hypothermia induced by either 30 minutes of total liquid ventilation (total liquid ventilation group, n = 12) or IV cold saline (conventional cooling group, n = 12).Measurements and Main Results: Ultrafast cooling with total liquid ventilation (32 degrees C within 5 min in the esophagus) dramatically attenuated the post-cardiac arrest syndrome regarding survival, neurologic dysfunction, and histologic lesions (brain, heart, kidneys, liver, and lungs). Final survival rate achieved 58% versus 0% and 8% in total liquid ventilation, control, and conventional cooling groups (p < 0.05), respectively. This was accompanied by an early preservation of the blood-brain barrier integrity and cerebral hemodynamics as well as reduction in the immediate reactive oxygen species production in the brain, heart, and kidneys after cardiac arrest. Later on, total liquid ventilation also mitigated the systemic inflammatory response through alteration of monocyte chemoattractant protein-1, interleukin-1, and interleukin-8 transcripts levels compared with control. In the conventional cooling group, cooling was achieved more slowly (32 degrees C within 90-120 min in the esophagus), providing none of the above-mentioned systemic or organ protection.Conclusions: Ultrafast cooling by total liquid ventilation limits the post-cardiac arrest syndrome after asphyxial cardiac arrest in rabbits. This protection involves an early limitation in reactive oxidative species production, blood-brain barrier disruption, and delayed preservation against the systemic inflammatory response.

OBJECTIVE:The protein Hwp1, expressed on the pathogenic phase of Candida albicans, presents sequence analogy with the gluten protein gliadin and is also a substrate for transglutaminase. This had led to the suggestion that C. albicans infection (CI) may be a triggering factor for Celiac disease (CeD) onset. We investigated cross-immune reactivity between CeD and CI.METHODS:Serum IgG levels against recombinant Hwp1 and serological markers of CeD were measured in 87 CeD patients, 41 CI patients, and 98 healthy controls (HC). IgA and IgG were also measured in 20 individuals from each of these groups using microchips sensitized with 38 peptides designed from the N-terminal of Hwp1.RESULTS:CI and CeD patients had higher levels of anti-Hwp1 (p=0.0005 and p=0.004) and anti-gliadin (p=0.002 and p=0.0009) antibodies than HC but there was no significant difference between CeD and CI patients. CeD and CI patients had higher levels of anti-transglutaminase IgA than HC (p=0.0001 and p=0.0039). During CI, the increase in anti-Hwp1 paralleled the increase in anti-gliadin antibodies. Microchip analysis showed that CeD patients were more reactive against some Hwp1 peptides than CI patients, and that some deamidated peptides were more reactive than their native analogs. Binding of IgG from CeD patients to Hwp1 peptides was inhibited by γIII gliadin peptides.CONCLUSIONS:Humoral cross-reactivity between Hwp1 and gliadin was observed during CeD and CI. Increased reactivity to Hwp1 deamidated peptide suggests that transglutaminase is involved in this interplay. These results support the hypothesis that CI may trigger CeD onset in genetically-susceptible individuals.

Objectives: Total liquid ventilation provides ultrafast and potently neuro- and cardioprotective cooling after shockable cardiac arrest and myocardial infarction in animals. Our goal was to decipher the effect of hypothermic total liquid ventilation on the systemic and cerebral response to asphyxial cardiac arrest using an original pressure- and volume-controlled ventilation strategy in rabbits.Design: Randomized animal study.Setting: Academic research laboratory.Subjects: New Zealand Rabbits.Interventions: Thirty-six rabbits were submitted to 13 minutes of asphyxia, leading to cardiac arrest. After resumption of spontaneous circulation, they underwent either normothermic life support (control group, n = 12) or hypothermia induced by either 30 minutes of total liquid ventilation (total liquid ventilation group, n = 12) or IV cold saline (conventional cooling group, n = 12).Measurements and Main Results: Ultrafast cooling with total liquid ventilation (32 degrees C within 5 min in the esophagus) dramatically attenuated the post-cardiac arrest syndrome regarding survival, neurologic dysfunction, and histologic lesions (brain, heart, kidneys, liver, and lungs). Final survival rate achieved 58% versus 0% and 8% in total liquid ventilation, control, and conventional cooling groups (p < 0.05), respectively. This was accompanied by an early preservation of the blood-brain barrier integrity and cerebral hemodynamics as well as reduction in the immediate reactive oxygen species production in the brain, heart, and kidneys after cardiac arrest. Later on, total liquid ventilation also mitigated the systemic inflammatory response through alteration of monocyte chemoattractant protein-1, interleukin-1, and interleukin-8 transcripts levels compared with control. In the conventional cooling group, cooling was achieved more slowly (32 degrees C within 90-120 min in the esophagus), providing none of the above-mentioned systemic or organ protection.Conclusions: Ultrafast cooling by total liquid ventilation limits the post-cardiac arrest syndrome after asphyxial cardiac arrest in rabbits. This protection involves an early limitation in reactive oxidative species production, blood-brain barrier disruption, and delayed preservation against the systemic inflammatory response.

Purpose of the study: Ultrafast cooling with total liquid ventilation (TLV) can induce a potent benefit in animal models of shockable cardiac arrest. Our purpose was to determine whether it can also induce neuroprotection after non shockable cardiac arrest.

Scedosporium boydii is an opportunistic filamentous fungus which may be responsible for a wide variety of infections in immunocompetent and immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex, which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF) and may lead to allergic bronchopulmonary mycoses, sensitization, or respiratory infections. Upon microbial infection, host phagocytic cells release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by detoxification of the hydrogen peroxide. Here, we investigated the catalase equipment of Scedosporium boydii, one of the major pathogenic species in the S. apiospermum species complex. Three catalases were identified, and the mycelial catalase A1 was purified to homogeneity by a three-step chromatographic process. This enzyme is a monofunctional tetrameric protein of 460 kDa, consisting of four 82-kDa glycosylated subunits. The potential usefulness of this enzyme in serodiagnosis of S. apiospermum infections was then investigated by an enzyme-linked immunosorbent assay (ELISA), using 64 serum samples from CF patients. Whatever the species involved in the S. apiospermum complex, sera from infected patients were clearly differentiated from sera from patients with an Aspergillus fumigatus infection or those from CF patients without clinical and biological signs of a fungal infection and without any fungus recovered from sputum samples. These results suggest that catalase A1 is a good candidate for the development of an immunoassay for serodiagnosis of infections caused by the S. apiospermum complex in patients with CF.

Introduction: We previously demonstrated that total liquid ventilation (TLV) with perfluorocarbons can afford ultra-fast cooling and improve outcome after non-shockable cardiac arrest in rabbits. Hypothesis: We determined whether this protection could be linked to an early inhibition of reperfusion injury after cardiac arrest, including oxidative stress and blood brain barrier (BBB) disruption. Methods: Anesthetized rabbits were randomly submitted to 13 min of asphyxia followed by cardiopulmonary resuscitation. Then, they randomly underwent normothermic life support (Control group) or hypothermia induced by either i.v. cold saline (30 ml/kg; CONV group) or TLV (TLV group). In a first set of experiments, reactive oxygen species (ROS) were measured by electron paramagnetic resonance spectroscopy using CMH as spin probe on tissues sampled 30 min after cardiac arrest. In a second set of experiments, BBB integrity was assessed by the administration of Evans blue dye (EBD) 30 min after resuscitation. Brains wer...

BACKGROUND:Species of the Scedosporium apiospermum complex (S. a complex) are emerging fungi responsible for chronic airway colonization in cystic fibrosis (CF) patients. Recent studies performed on Aspergillus fumigatus suggest that the colonization of the airways by filamentous fungi may contribute to the progressive deterioration of lung function.METHODS:We studied S. a complex seroprevalence, as a marker of close contact between patient and the fungi, in a large monocentric cohort of CF patients attended in a reference centre in Lyon, France.RESULTS:Serum samples from 373 CF patients were analysed. Antibodies against S. a complex were detected in 35 patients (9.4%). In multivariate analysis, S. a complex seropositivity was only associated with seropositivity to A. fumigatus.CONCLUSIONS:This study does not suggest an association between sensitization against S. a complex and poorer lung function in CF. Prospective studies are needed to evaluate the impact of both seropositivity and S. a complex colonization on the course of CF.

Purpose of the study: Total liquid ventilation (TLV) provides ultrafast cooling and potent anti-ischemic effects in animal models of myocardial infarction and cardiac arrest. We determined whether it could also mitigate shock in an original model of multiorgan failure induced by abdominal ischemia-reperfusion.

In total liquid ventilation (TLV), the lungs are filled with a breathable liquid perfluorocarbon (PFC) while a liquid ventilator ensures proper gas exchange by renewal of a tidal volume of oxygenated and temperature-controlled PFC. Given the rapid changes in core body temperature generated by TLV using the lung has a heat exchanger, it is crucial to have accurate and reliable core body temperature monitoring and control. This study presents the design of a virtual lung temperature sensor to control core temperature. In the first step, the virtual sensor, using expired PFC to estimate lung temperature noninvasively, was validated both in vitro and in vivo. The virtual lung temperature was then used to rapidly and automatically control core temperature. Experimentations were performed using the Inolivent-5.0 liquid ventilator with a feedback controller to modulate inspired PFC temperature thereby controlling lung temperature. The in vivo experimental protocol was conducted on seven newborn lambs instrumented with temperature sensors at the femoral artery, pulmonary artery, oesophagus, right ear drum, and rectum. After stabilization in conventional mechanical ventilation, TLV was initiated with fast hypothermia induction, followed by slow posthypothermic rewarming for 1 h, then by fast rewarming to normothermia and finally a second fast hypothermia induction phase. Results showed that the virtual lung temperature was able to provide an accurate estimation of systemic arterial temperature. Results also demonstrate that TLV can precisely control core body temperature and can be favorably compared to extracorporeal circulation in terms of speed.

Total liquid ventilation is an innovative experimental method of mechanical assisted ventilation in which lungs are totally filled and then ventilated with a tidal volume of perfluorochemical liquid (PFC) by using a dedicated liquid ventilator. The positive end-inspiratory and end-expiratory pressures (PEIP and PEEP) are static pressure measurements that are critic to the safe and efficient control of the ventilation. However, their measurement is impeded by large oscillations of pressure caused by the propagation of pressure waves along the flexible tubes carrying the PFC to the patient. The aim of this paper is to describe a method to accurately estimate the PEEP and the PEIP from noisy data hindered by flexible tubing resonance during short respiratory pauses. The method developped makes use of the least squares technique to estimate the steady state pressure. Preliminary in vivo validation of the algorithm shows that the method gives accurate estimations with respiratory pauses as short as 0.3 second.