Total liquid ventilation in an ovine model of extreme prematurity: a randomized study.

BACKGROUND: This study aimed at comparing cardiorespiratory stability during total liquid ventilation (TLV)-prior to lung aeration-with conventional mechanical ventilation (CMV) in extremely preterm lambs during the first 6 h of life. METHODS: 23 lambs (11 females) were born by c-section at 118-120 days of gestational age (term = 147 days) to receive 6 h of TLV or CMV from birth. Lung samples were collected for RNA and histology analyses. RESULTS: The lambs under TLV had higher and more stable arterial oxygen saturation (p = 0.001) and cerebral tissue oxygenation (p = 0.02) than the lambs in the CMV group in the first 10 min of transition to extrauterine life. Although histological assessment of the lungs was similar between the groups, a significant upregulation of IL-1a, IL-6 and IL-8 RNA in the lungs was observed after TLV. CONCLUSIONS: Total liquid ventilation allowed for remarkably stable transition to extrauterine life in an extremely preterm lamb model. Refinement of our TLV prototype and ventilation algorithms is underway to address specific challenges in this population, such as minimizing tracheal deformation during the active expiration. IMPACT: Total liquid ventilation allows for remarkably stable transition to extrauterine life in an extremely preterm lamb model. Total liquid ventilation is systematically achievable over the first 6 h of life in the extremely premature lamb model. This study provides additional incentive to pursue further investigation of total liquid ventilation as a transition tool for the most extreme preterm neonates.

Efficacy and Safety of Eculizumab in Pediatric Patients Affected by Shiga Toxin-Related Hemolytic and Uremic Syndrome: A Randomized, Placebo-Controlled Trial.

SIGNIFICANCE STATEMENT: Shiga toxin-related hemolytic uremic syndrome (STEC-HUS) is a serious condition, characterized by multiorgan thrombotic microangiopathy, mainly affecting children. Renal involvement is severe, with approximately half of patients requiring dialysis. So far, no specific treatment has been proven efficient in STEC-HUS. The use of eculizumab, a monoclonal antibody inhibiting terminal complement complex, has demonstrated remarkable success in atypical hemolytic uremic syndrome, but its use in uncontrolled studies to treat STEC-HUS has yielded inconsistent results. In this Phase 3 randomized, placebo-controlled trial in 100 pediatric patients with STEC-HUS, the findings did not show efficacy of eculizumab during the acute phase of the disease. However, the results indicated a reduction of renal sequelae in eculizumab-treated patients at 1-year follow-up. Larger prospective studies would be needed to further explore eculizumab as a potential treatment. BACKGROUND: Shiga toxin-related hemolytic uremic syndrome (STEC-HUS) in children is a severe condition, resulting in approximately 50% of patients requiring RRT. Furthermore, at least 30% of survivors experience kidney sequelae. Recently, activation of the complement alternative pathway has been postulated as a factor in STEC-HUS pathophysiology, leading to compassionate use of eculizumab, a monoclonal antibody inhibiting the terminal complement complex, in affected patients. Given the lack of therapy for STEC-HUS, a controlled study of eculizumab efficacy in treating this condition is a priority. METHODS: We conducted a Phase 3 randomized trial of eculizumab in children with STEC-HUS. Patients were randomly assigned in a 1:1 ratio to receive either eculizumab or placebo during 4 weeks. Follow-up lasted for 1 year. The primary end point was RRT duration <48 hours after randomization. Secondary endpoints included hematologic and extrarenal involvement. RESULTS: Baseline characteristics were similar among the 100 patients who underwent randomization. The rate of RRT <48 hours did not differ significantly between the two groups (48% in the placebo versus 38% in the eculizumab group; P = 0.31) or in the course of ARF. The two groups also exhibited similar hematologic evolution and extrarenal manifestations of STEC-HUS. The proportion of patients experiencing renal sequelae at 1 year was lower in the eculizumab group than in the placebo group (43.48% and 64.44%, respectively, P = 0.04). No safety concern was reported. CONCLUSIONS: In pediatric patients with STEC-HUS, eculizumab treatment does not appear to be associated with improved renal outcome during acute phase of the disease but may reduce long-term kidney sequelae. CLINICAL TRIALS REGISTRATIONS: EUDRACT (2014-001169-28) ClinicalTrials.gov ( NCT02205541 ).

Hemodynamic impacts of apelin-13 in a neonatal lamb model of septic peritonitis.

BACKGROUND: Apelins are potential candidate therapeutic molecules for hemodynamic support. The objective of this study was to assess the hemodynamic impacts of apelin-13 in a neonatal lamb model of septic shock. METHODS: Lambs were randomized to receive apelin-13 or normal saline. Septic shock was induced by injecting a fecal slurry into the peritoneal cavity. Lambs underwent volume repletion (30 mL/kg over 1 h) followed by intravenous administration of 5 incremental doses (D) of apelin-13 (D1 = 0.039 to D5 = 19.5 µg/kg/h) or normal saline. RESULTS: Following fecal injection, mean arterial pressure (MAP) and cardiac index (CI) dropped in both groups (p < 0.05). The MAP decreased non-significantly from D1 to D5 (p = 0.12) in the saline group, while increasing significantly (p = 0.02) in the apelin group (-12 (-17; 12) vs. +15 (6; 23) % (p = 0.012)). Systemic vascular resistances were higher in the apelin-13 group at D5 compared to the saline group (4337 (3239, 5144) vs. 2532 (2286, 3966) mmHg/min/mL, respectively, p = 0.046). The CI increased non-significantly in the apelin-13 group. CONCLUSION: Apelin-13 increased MAP in a neonatal lamb septic shock model. IMPACT: Administration of apelin-13 stabilized hemodynamics during the progression of the sepsis induced in this neonatal lamb model. Systemic vascular resistances were higher in the apelin-13 group than in the placebo group. This suggests ontogenic differences in vascular response to apelin-13 and warrants further investigation. This study suggests that apelin-13 could eventually become a candidate for the treatment of neonatal septic shock.

Phytoremediation of PCB: contaminated Algerian soils using native agronomics plants.

Pot cultivation experiments were conducted to assess the phytoremediation potential of two local agronomic plants, namely Avena sativa and Vicia sativa. Several soils with long-standing contamination and different levels of Polychlorinated biphenyl (PCB) contamination were used for this study. The soil samples came from different regions of Algeria and had different physico-chemical parameters. We studied the influence of these parameters on remediation potential of the two tested plants. The removal rate of the seven PCBs (PCB 28, 52, 101, 138, 153, 156 and 180) was examined after 40 and 90 days. The results showed that the presence of the plants reduced significantly the overall PCB content, ranging initially from 1.33-127.9 mg kg1. After 90 days, the forage plant Vicia sativa allowed us to reach an excess dissipation rate of 56.7% compared to the unplanted control for the most polluted soil. An average dissipation rate of 50% was obtained in the moderately polluted soil. The less contaminated soil had an excess dissipation rate of about 24% for both plants and a predominant dissipation of the low chlorinated PCBs.

Development of a standardized in vitro approach to evaluate microphysical, chemical, and toxicological properties of combustion-derived fine and ultrafine particles.

Ultrafine particles represent a growing concern in the public health community but their precise role in many illnesses is still unknown. This lack of knowledge is related to the experimental difficulty in linking their biological effects to their multiple properties, which are important determinants of toxicity. Our aim is to propose an interdisciplinary approach to study fine (FP) and ultrafine (UFP) particles, generated in a controlled manner using a miniCAST (Combustion Aerosol Standard) soot generator used with two different operating conditions (CAST1 and CAST3). The chemical characterization was performed by an untargeted analysis using ultra-high resolution mass spectrometry. In conjunction with this approach, subsequent analysis by gas chromatography-mass spectrometry (GC-MS) was performed to identify polycyclic aromatic hydrocarbons (PAH). CAST1 enabled the generation of FP with a predominance of small PAH molecules, and CAST3 enabled the generation of UFP, which presented higher numbers of carbon atoms corresponding to larger PAH molecules. Healthy normal human bronchial epithelial (NHBE) cells differentiated at the air-liquid interface (ALI) were directly exposed to these freshly emitted FP and UFP. Expression of MUC5AC, FOXJ1, OCLN and ZOI as well as microscopic observation confirmed the ciliated pseudostratified epithelial phenotype. Study of the mass deposition efficiency revealed a difference between the two operating conditions, probably due to the morphological differences between the two categories of particles. We demonstrated that only NHBE cells exposed to CAST3 particles induced upregulation in the gene expression of IL-8 and NQO1. This approach offers new perspectives to study FP and UFP with stable and controlled properties.

Comprehensive analysis of a widely pharmaceutical, furosemide, and its degradation products in aquatic systems: Occurrence, fate, and ecotoxicity.

Many pharmaceutical compounds end up in the environment due to incomplete removal by wastewater treatment plants (WWTPs). Some compounds are sometimes present in significant concentrations and therefore represent a risk to the aquatic environment. Furosemide is one of the most widely used drugs in the world. Considered as an essential drug by the World Health Organization, this powerful loop diuretic is used extensively to treat hypertension, heart and kidney failure and many other purposes. However, this important consumption also results in a significant release of furosemide in wastewater and in the receiving environment where concentrations of a few hundred ng/L to several thousand have been found in the literature, making furosemide a compound of great concern. Also, during its transport in wastewater systems and WWTPs, furosemide can be degraded by various processes resulting in the production of more than 74 by-products. Furosemide may therefore present a significant risk to ecosystem health due not only to its direct cytotoxic, genotoxic and hepatotoxic effects in animals, but also indirectly through its transformation products, which are poorly characterized. Many articles classify furosemide as a priority pollutant according to its occurrence in the environment, its persistence, its elimination by WWTPs, its toxicity and ecotoxicity. Here, we present a state-of-the-art review of this emerging pollutant of interest, tracking it, from its consumption to its fate in the aquatic environment. Discussion points include the occurrence of furosemide in various matrices, the efficiency of many processes for the degradation of furosemide, the subsequent production of degradation products following these treatments, as well as their toxicity.

Measuring aliphatic hydrocarbons in sediments by direct thermal desorption-gas chromatography-mass spectrometry: Matrix effects and quantification challenges.

Direct sample introduction thermal desorption (TD) coupled to GC-MS was investigated for the analysis of paraffinic hydrocarbons (HCs) from polluted sediments. TD-GC-MS is sometimes used for analysing paraffinic HCs from atmospheric particles but rarely for their direct desorption from sediments. So, the new TD methodology, applied to sediments, required development, optimization and validation. A definitive screening experimental design was performed to discriminate the critical factors on TD efficiency, from model sediments containing various organic matter (OM) amounts. Low molecular weight HCs had extraction behaviours markedly different from high molecular ones (HMW-HCs), but a compromise was found using very few sediment amount (5 mg), high temperature rate (55 °C min-1) and final temperature (350 °C). Linear HCs (n-C10 to n-C40) could be quantified using the matrix-matched calibration method, with very low detection limits (3.8-13.4 ng). The amount of the overall paraffinic alkanes was also determined as a sum of unresolved components between predefined equivalent carbon ranges. The developed solventless methodology was compared to an optimized solvent microwave assisted extraction (MAE). Matrix effects could be higher for TD compared to MAE but it depended on sediment matrix. When matrix effect was strong, particularly on HMW-HCs signal depletion, a dilution with pure non-porous sand was favourable for accurate quantification. The sum of resolved and unresolved HCs gave comparable results between MAE and TD extractions, with an exception of alkanes greater than C30 which were less quantitatively extracted via TD. However, TD-GC-MS was more sensitive than MAE-GC-MS. So TD-GC-MS is useful for analyzing sediments containing a great range of paraffinic HCs (C9-C34) and it has the advantages of being fully automated, with few sample preparation and operator intervention, using very low amounts of solvent, and generating few wastes.

Effect of body position during weaning from total liquid ventilation in piglets.

OBJECTIVE: To determine if change in body position improves oxygen requirements and respiratory mechanics during the transition from total liquid ventilation (TLV) to gas ventilation. METHODS: Fourteen piglets underwent TLV, followed by a 2-hour weaning period under conventional gas ventilation. Subjects were randomized to the experimental group (Rotating - R), that was in prone position between the 10th and 30th minute of weaning, or to the static control group (Supine - S). RESULTS: Oxygenation index was lower in the R group at 30 minutes in prone position than that in the S group (1.9 [1.6; 2.8] vs 3.5 [3.1; 5.1], p = 0.001). This difference disappeared when subjects resumed the supine position (4.2 [3.8; 4.7] and 4.7 [3.8; 5.4], p = 0.4, for the R and S groups, respectively). The change in body position did not affect respiratory system compliance or inspiratory capacity. CONCLUSION: Prone position improved oxygenation during weaning from TLV. The effect disappeared once piglets returned to the supine position.

Age-related changes in rat myocardium involve altered capacities of glycosaminoglycans to potentiate growth factor functions and heparan sulfate-altered sulfation.

Glycosaminoglycans (GAGs) are essential components of the extracellular matrix, the natural environment from which cell behavior is regulated by a number or tissue homeostasis guarantors including growth factors. Because most heparin-binding growth factor activities are regulated by GAGs, structural and functional alterations of these polysaccharides may consequently affect the integrity of tissues during critical physiological and pathological processes. Here, we investigated whether the aging process can induce changes in the myocardial GAG composition in rats and whether these changes can affect the activities of particular heparin-binding growth factors known to sustain cardiac tissue integrity. Our results showed an age-dependent increase of GAG levels in the left ventricle. Biochemical and immunohistological studies pointed out heparan sulfates (HS) as the GAG species that increased with age. ELISA-based competition assays showed altered capacities of the aged myocardial GAGs to bind FGF-1, FGF-2, and VEGF but not HB EGF. Mitogenic assays in cultured cells showed an age-dependent decrease of the elderly GAG capacities to potentiate FGF-2 whereas the potentiating effect on VEGF(165) was increased, as confirmed by augmented angiogenic cell proliferation in Matrigel plugs. Moreover, HS disaccharide analysis showed considerably altered 6-O-sulfation with modest changes in N- and 2-O-sulfations. Together, these findings suggest a physiological significance of HS structural and functional alterations during aging. This can be associated with an age-dependent decline of the extracellular matrix capacity to efficiently modulate not only the activity of resident or therapeutic growth factors but also the homing of resident or therapeutic cells.

Self-evolving oxidative stress with identifiable pre- and postmitochondrial phases in PC12 cells.

During the neurodegenerative process in several brain diseases, oxidative stress is known to play important roles in disease severity and evolution. Although early events of stress, such as increased lipid peroxidation and decreased superoxide dismutase, are known to characterize early onsets of these diseases, little is known about the events that participate in maintaining the chronic evolving phase influencing the disease progression in neurons. Here, we used differentiated PC12 cells to identify premitochondrial and postmitochondrial events occurring during the oxidative stress cascade leading to apoptosis. Our data indicate that an acute and strong oxidative impulse (500 µM H(2)O(2), 30 min) can induce, in this model, a 24-hr self-evolving stress, which advances from a premitochondrial phase characterized by lysosomes and cathepsin B and D translocations to cytosol and early mitochondrial membrane hyperpolarization. This phase lasts for about 5 hr and is followed by a postmitochondrial phase distinguished by mitochondrial membrane depolarization, reactive oxygen species increase, caspase-9 and caspase-3 activations, and apoptosis. Inhibition of cathepsins B and D suggests that cells can be protected at the premitochondrial phase of stress evolution and that new cathepsins regulators, such as glycosaminoglycans mimetics, can be considered as new therapeutic prototypes for neurodegeneration. Insofar as early oxidative stress markers have been related to the early onset of neurodegeneration, strategies protecting cells at the premitochondrial phase of oxidative stress may have important therapeutic applications.

Occurrence and fate of an emerging drug pollutant and its by-products during conventional and advanced wastewater treatment: Case study of furosemide.

Conventional wastewater treatment systems are not designed to remove pharmaceutical compounds from wastewater. These compounds can be degraded into many other transformation products which are hardly, if at all, studied. In this context, we studied the occurrence and degradation of furosemide, a very frequently detected diuretic, along with its known degradation products in several types of wastewater. Influent and effluent from the Seine-Centre Wastewater Treatment Plant (WWTP) (Paris, France) as well as outlet of residential care homes (Dordogne, France) were analyzed by Ultra-Performance Liquid Chromatography-tandem Mass Spectrometry (UPLC-MS/MS) to quantify furosemide and its known degradation products, saluamine and pyridinium of furosemide. Oxidation experiments (chlorination, ozonation and UV photolysis with hydrogen peroxide) were then performed on furosemide solutions and on water from residential care facilities to study the degradation of furosemide by potential advanced processes, and also to identify unknown oxidation products by high-resolution mass spectrometry. Furosemide was well degraded in Seine-Centre WWTP (>75%) but did not increase the concentrations of its main degradation products. Saluamine and pyridinium of furosemide were already present at similar concentrations to furosemide in the raw wastewater (∼2.5-3.5 µg.L-1), and their removal in the WWTPs were very high (>80%). Despite their removal, the three compounds remained present in treated wastewater effluents at concentrations of hundreds of nanograms per liter. Chlorination degraded furosemide without pyridinium production unlike the other two processes. Chlorination and ozonation were also effective for the removal of furosemide and pyridinium in residential care home water, but they resulted in the production of saluamine. To our knowledge this is the first evidence of saluamine and pyridinium of furosemide in real water samples in either the particulate or dissolved phase.

A Drug-Eluting Injectable NanoGel for Localized Delivery of Anticancer Drugs to Solid Tumors.

Systemically administered chemotherapy reduces the efficiency of the anticancer agent at the target tumor tissue and results in distributed drug to non-target organs, inducing negative side effects commonly associated with chemotherapy and necessitating repeated administration. Injectable hydrogels present themselves as a potential platform for non-invasive local delivery vehicles that can serve as a slow-releasing drug depot that fills tumor vasculature, tissue, or resection cavities. Herein, we have systematically formulated and tested an injectable shear-thinning hydrogel (STH) with a highly manipulable release profile for delivering doxorubicin, a common chemotherapeutic. By detailed characterization of the STH physical properties and degradation and release dynamics, we selected top candidates for testing in cancer models of increasing biomimicry. Two-dimensional cell culture, tumor-on-a-chip, and small animal models were used to demonstrate the high anticancer potential and reduced systemic toxicity of the STH that exhibits long-term (up to 80 days) doxorubicin release profiles for treatment of breast cancer and glioblastoma. The drug-loaded STH injected into tumor tissue was shown to increase overall survival in breast tumor- and glioblastoma-bearing animal models by 50% for 22 days and 25% for 52 days, respectively, showing high potential for localized, less frequent treatment of oncologic disease with reduced dosage requirements.

Modulation of inflammation by Cicaderma ointment accelerates skin wound healing.

Skin wound healing is a natural and intricate process that takes place after injury, involving different sequential phases such as hemostasis, inflammatory phase, proliferative phase, and remodeling that are associated with complex biochemical events. The interruption or failure of wound healing leads to chronic nonhealing wounds or fibrosis-associated diseases constituting a major health problem where, unfortunately, medicines are not very effective. The objective of this study was to evaluate the capacity of Cicaderma ointment (Boiron, Lyon, France) to accelerate ulcer closure without fibrosis and investigate wound healing dynamic processes. We used a necrotic ulcer model in mice induced by intradermal doxorubicin injection, and after 11 days, when the ulcer area was maximal, we applied Vaseline petroleum jelly or Cicaderma every 2 days. Topical application of Cicaderma allowed a rapid recovery of mature epidermal structure, a more compact and organized dermis and collagen bundles compared with the Vaseline group. Furthermore, the expression of numerous cytokines/molecules in the ulcer was increased 11 days after doxorubicin injection compared with healthy skin. Cicaderma rapidly reduced the level of proinflammatory cytokines, mainly tumor necrosis factor (TNF)-α and others of the TNF pathway, which can be correlated to a decrease of polymorphonuclear recruitment. It is noteworthy that the modulation of inflammation through TNF-α, macrophage inflammatory protein-1α, interleukin (IL)-12, IL-4, and macrophage-colony-stimulating factor was maintained 9 days after the first ointment application, facilitating the wound closure without affecting angiogenesis. These cytokines seem to be potential targets for therapeutic approaches in chronic wounds. Our results confirm the use of Cicaderma for accelerating skin wound healing and open new avenues for sequential treatments to improve healing.

Direct thermal desorption-gas chromatography-tandem mass spectrometry versus microwave assisted extraction and GC-MS for the simultaneous analysis of polyaromatic hydrocarbons (PAHs, PCBs) from sediments.

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) are regulated contaminants usually investigated in sediments. Conventional approaches often use GC-MS to analyse them with a preliminary extraction step which can be solvent- and time-consuming. Here two extraction methodologies were optimized using experimental designs, and compared: microwave assisted extraction (MAE) and thermal desorption (TD); the latter was rarely used for sediments analyses. Several factors that may influence extraction recoveries were studied including matrix parameters (mass, organic matter (OM) content) and processing parameters. A definitive screening design DSD was performed to screen the 6 most influencing factors and model the extraction recoveries using TD. Whatever the OM content, a minimum sediment mass (5 mg) was better for an optimal extraction, with a minimum temperature rate (15 °C min-1), a maximum final temperature (350 °C) associated with a minimum hold time (5 min), and a maximum vent flow (150 mL min-1) between the TD unit and the cryogenic trap. Thereafter matrix effects were evaluated using standard addition, and quality assurance and control were implemented for comparing MAE and TD. TD-GC-MS/MS sensitivity was higher than MAE-GC-MS with detection limits in the range 5-1160 pg and 20-125 pg for PAHs and PCBs, respectively. When considering the appropriate strategy for quantification, TD was also reliable for sediments analysis. Although MAE was less sensitive to matrix effects, TD could significantly improve the analytical process, due to direct coupling with GC-MS/MS and complete automation. Moreover, TD offered possible higher spatial resolution than MAE, particularly for sediment cores analysis, due to the 1000-times lower sample size. At last, TD-GC-MS/MS appeared as a greener analytical procedure.

Conventional vs high-frequency ventilation for weaning from total liquid ventilation in lambs.

OBJECTIVE: To compare conventional gas ventilation (GV) and high-frequency oscillatory ventilation (HFOV) for weaning from total liquid ventilation (TLV). METHODS: Sixteen lambs were anesthetized. After 1 h of TLV with perflubron (PFOB), they were assigned to either GV or HFOV for 2 h. Oxygen requirements, electrical impedance tomography and videofluoroscopic sequences, and respiratory system compliance were recorded. RESULTS: The lambs under GV needed less oxygen at 20 min following TLV (40 [25, 45] and 83 [63, 98]%, p = 0.001 under GV and HFOV, respectively). During weaning, tidal volume distribution was increased in the nondependent regions in the GV group compared to baseline (p = 0.046). Furthermore, residual PFOB was observed in the most dependent region. No air was detected by fluoroscopy in that region at the end of expiration in the GV group. CONCLUSION: GV offers a transient advantage over HFOV with regards to oxygenation for TLV weaning.

Ultrafine Particles Issued from Gasoline-Fuels and Biofuel Surrogates Combustion: A Comparative Study of the Physicochemical and In Vitro Toxicological Effects.

Gasoline emissions contain high levels of pollutants, including particulate matter (PM), which are associated with several health outcomes. Moreover, due to the depletion of fossil fuels, biofuels represent an attractive alternative, particularly second-generation biofuels (B2G) derived from lignocellulosic biomass. Unfortunately, compared to the abundant literature on diesel and gasoline emissions, relatively few studies are devoted to alternative fuels and their health effects. This study aimed to compare the adverse effects of gasoline and B2G emissions on human bronchial epithelial cells. We characterized the emissions generated by propane combustion (CAST1), gasoline Surrogate, and B2G consisting of Surrogate blended with anisole (10%) (S+10A) or ethanol (10%) (S+10E). To study the cellular effects, BEAS-2B cells were cultured at air-liquid interface for seven days and exposed to different emissions. Cell viability, oxidative stress, inflammation, and xenobiotic metabolism were measured. mRNA expression analysis was significantly modified by the Surrogate S+10A and S+10E emissions, especially CYP1A1 and CYP1B1. Inflammation markers, IL-6 and IL-8, were mainly downregulated doubtless due to the PAHs content on PM. Overall, these results demonstrated that ultrafine particles generated from biofuels Surrogates had a toxic effect at least similar to that observed with a gasoline substitute (Surrogate), involving probably different toxicity pathways.

Life cycle studies of the hexose transporter of Plasmodium species and genetic validation of their essentiality.

A Plasmodium falciparum hexose transporter (PfHT) has previously been shown to be a facilitative glucose and fructose transporter. Its expression in Xenopus laevis oocytes and the use of a glucose analogue inhibitor permitted chemical validation of PfHT as a novel drug target. Following recent re-annotations of the P. falciparum genome, other putative sugar transporters have been identified. To investigate further if PfHT is the key supplier of hexose to P. falciparum and to extend studies to different stages of Plasmodium spp., we functionally analysed the hexose transporters of both the human parasite P. falciparum and the rodent parasite Plasmodium berghei using gene targeting strategies. We show here the essential function of pfht for the erythrocytic parasite growth as it was not possible to knockout pfht unless the gene was complemented by an episomal construct. Also, we show that parasites are rescued from the toxic effect of a glucose analogue inhibitor when pfht is overexpressed in these transfectants. We found that the rodent malaria parasite orthologue, P. berghei hexose transporter (PbHT) gene, was similarly refractory to knockout attempts. However, using a single cross-over transfection strategy, we generated transgenic P. berghei parasites expressing a PbHT-GFP fusion protein suggesting that locus is amenable for gene targeting. Analysis of pbht-gfp transgenic parasites showed that PbHT is constitutively expressed through all the stages in the mosquito host in addition to asexual stages. These results provide genetic support for prioritizing PfHT as a target for novel antimalarials that can inhibit glucose uptake and kill parasites, as well as unveiling the expression of this hexose transporter in mosquito stages of the parasite, where it is also likely to be critical for survival.

Use of a selective inhibitor to define the chemotherapeutic potential of the plasmodial hexose transporter in different stages of the parasite's life cycle.

During blood infection, malarial parasites use D-glucose as their main energy source. The Plasmodium falciparum hexose transporter (PfHT), which mediates the uptake of D-glucose into parasites, is essential for survival of asexual blood-stage parasites. Recently, genetic studies in the rodent malaria model, Plasmodium berghei, found that the orthologous hexose transporter (PbHT) is expressed throughout the parasite's development within the mosquito vector, in addition to being essential during intraerythrocytic development. Here, using a D-glucose-derived specific inhibitor of plasmodial hexose transporters, compound 3361, we have investigated the importance of D-glucose uptake during liver and transmission stages of P. berghei. Initially, we confirmed the expression of PbHT during liver stage development, using a green fluorescent protein (GFP) tagging strategy. Compound 3361 inhibited liver-stage parasite development, with a 50% inhibitory concentration (IC50) of 11 µM. This process was insensitive to the external D-glucose concentration. In addition, compound 3361 inhibited ookinete development and microgametogenesis, with IC50s in the region of 250 µM (the latter in a D-glucose-sensitive manner). Consistent with our findings for the effect of compound 3361 on vector parasite stages, 1 mM compound 3361 demonstrated transmission blocking activity. These data indicate that novel chemotherapeutic interventions that target PfHT may be active against liver and, to a lesser extent, transmission stages, in addition to blood stages.

Air distribution within the lungs after total liquid ventilation in a neonatal ovine model.

OBJECTIVE: To gain insight into the total and regional lung aeration dynamics at the transition from total liquid ventilation (TLV) to conventional mechanical ventilation (GV). METHODS: Neonatal lambs received either TLV for 4 h followed by GV (n = 15) or GV only (n = 11, controls). Monitoring was performed in the prone position with both videofluoroscopy and electrical impedance tomography (EIT) for the first 10 min of the transition. RESULTS: Total and regional end-expiratory lung volumes were stable throughout the transition (p < 0.05). The percentage of tidal volume, liquid and/or gaseous, distributed to the different regions was stable (p < 0.05). Radiopacity of the nondependent regions markedly decreased at end-expiration (p < 0.01), reflecting the progressive transition to a gaseous end-expiratory lung volume. CONCLUSION: Weaning to GV did not increase total or regional lung volumes, suggesting that the risk of overdistention was not increased. Residual perfluorocarbon in the dependent lung regions might account for the high O2 needs we observed in the first minutes of GV after TLV.

Design and synthesis of a cyclitol-derived scaffold with axial pyridyl appendages and its encapsulation of the silver(I) cation.

Conversion of a myo-inositol derivative into a scyllo-inositol-derived scaffold with C3(v) symmetry bearing three axial pyridyl appendages is presented. This pre-organized hexadentate ligand allows complexation of silver(I). The crystal structure of the complex was established.