Comparative evaluation of FUNGITEST and broth microdilution methods for antifungal drug susceptibility testing of Candida species and Cryptococcus neoformans

The FUNGITEST method (Sanofi Diagnostics Pasteur, Paris, France) is a microplate-based procedure for the breakpoint testing of six antifungal agents (amphotericin B, flucytosine, fluconazole, itraconazole, ketoconazole, and miconazole). We compared the FUNGITEST method with a broth microdilution test, performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines, for determining the in vitro susceptibilities of 180 isolates of Candida spp. (50 C. albicans, 50 C. glabrata, 10 C. kefyr, 20 C. krusei, 10 C. lusitaniae, 20 C. parapsilosis, and 20 C. tropicalis isolates) and 20 isolates of Cryptococcus neoformans. Overall, there was 100% agreement between the methods for amphotericin B, 95% agreement for flucytosine, 84% agreement for miconazole, 83% agreement for itraconazole, 77% agreement for ketoconazole, and 76% agreement for fluconazole. The overall agreement between the methods exceeded 80% for all species tested with the exception of C. glabrata (71% agreement). The poorest agreement between the results for individual agents was seen with C. glabrata (38% for fluconazole, 44% for ketoconazole, and 56% for itraconazole) and C. tropicalis (50% for miconazole). The FUNGITEST method misclassified as susceptible 2 of 12 (16.6%) fluconazole-resistant isolates, 2 of 10 (20%) itraconazole-resistant isolates, and 4 of 8 (50%) ketoconazole-resistant isolates of several Candida spp. Further development of the FUNGITEST procedure will be required before it can be recommended as an alternative method for the susceptibility testing of Candida spp. or C. neoformans.     

Strong dependence on pressure of the performance of a Co/SiO2 catalyst in Fischer–Tropsch slurry reactor synthesis

A 10 wt% Co/SiO₂ catalyst was prepared by the incipient wet-impregnation method and tested in Fischer–Tropsch synthesis in a slurry reactor under conditions approaching industrial practice. The catalyst precursor was investigated by X-ray diffraction (XRD), temperature-programmed reduction (TPR), transmission electron microscopy (TEM) and photoelectron spectroscopy (XPS). The XPS and XRD techniques revealed the presence of a crystalline Co₃O₄ spinel-type phase, while-in addition-TEM and XPS analyses pointed to the formation of another amorphous Co₃O₄ spinel phase, both species interacting weakly with the silica substrate. The influence of total pressure on the conversion, selectivity and stability of the catalyst was studied. Upon increasing the overall pressure from 20 to 40 bar, not only activity increased but also the catalyst are not deactivating. These results are explained in terms of an increase of gases solubility in the solvent, this increment of CO concentration in the liquid phase favours carbonyl species formation and the cobalt particles segregation that implies an increase in the metal surface area.     

Inhibition of neuronal calcium channels by a novel peptide spider toxin, DW13.3

Peptide toxins have proved to be useful agents, both in discriminating between different components of native calcium channel currents and in the molecular isolation and designation of their cloned channel counterparts. Here, we describe the isolation and characterization of the biochemical and physiological properties of a novel 74-amino acid peptide toxin (DW13.3) extracted from the venom of the spider Filistata hibernalis. The subtype specificity of DW13.3 was investigated using calcium channel currents recorded from two separate expression systems and several different cultured mammalian cell preparations. Overall, DW13.3 potently blocked all native calcium channel currents studied, with the exception of T-type currents recorded from GH3 cells. Examination of transiently expressed calcium channels in oocytes showed that DW13.3 had the highest affinity for alpha1A, followed by alpha1B > alpha1C > alpha1E. The affinity of DW13.3 for alpha1B N-type currents varied by 10-fold between expressed channels and native currents. Although block occurred in a similar 1:1 manner for all subtypes, DW13.3 produced a partial block of both alpha1A currents and P-type currents in cerebellar Purkinje cells. Selective occlusion of the P/Q-type channel ligand omega-conotoxin MVIIC (but not omega-agatoxin IVA) from its binding site in Purkinje neurons suggests that DW13.3 binds to a site close to the pore of the channel. The inhibition of different subtypes of calcium channels by DW13.3 reflects a common "macro" binding site present on all calcium channels except T-type.     

Community of inquiry as a theoretical framework to foster “epistemic engagement” and “cognitive presence” in online education

In this paper, several recent theoretical conceptions of technology-mediated education are examined and a study of 2159 online learners is presented. The study validates an instrument designed to measure teaching, social, and cognitive presence indicative of a community of learners within the community of inquiry (CoI) framework [Garrison, D. R., Anderson, T., & Archer, W. (2000). Critical inquiry in a text-based environment: Computer conferencing in higher education. The Internet and Higher Education, 2, 1–19; Garrison, D. R., Anderson, T., & Archer, W. (2001). Critical thinking, cognitive presence, and computer conferencing in distance education. American Journal of Distance Education, 15(1), 7–23]. Results indicate that the survey items cohere into interpretable factors that represent the intended constructs. Further it was determined through structural equation modeling that 70% of the variance in the online students’ levels of cognitive presence, a multivariate measure of learning, can be modeled based on their reports of their instructors’ skills in fostering teaching presence and their own abilities to establish a sense of social presence. Additional analysis identifies more details of the relationship between learner understandings of teaching and social presence and its impact on their cognitive presence. Implications for online teaching, policy, and faculty development are discussed.     

Synthetic substrates for human factor VIIa and factor VIIa-tissue factor

A series of 100 tripeptide fluorogenic substrates has been synthesized. These substrates contain Arg in the P1 position, various amino acids in the P2 and P3 positions, and different 6-amino-1-naphthalenesulfonamides (ANSN) as the detecting group (P'). The 38 compounds possessing the highest initial rates of factor VIIa hydrolysis were evaluated for substrate kinetic parameters in the presence and absence of tissue factor (TF) and by factor Xa. Most of these substrates had a higher kcat/KM (keff) value for the factor VIIa-TF complex than for factor Xa. Substitution of different amino acids in the P2 position showed that substrates with bulkier amino acids such as Leu, Pro, and Val have higher values for KM and kcat than those with smaller amino acids (Gly or Ser). The highest second-order rate constants were found for substrates with Val or Pro in the P2 position. A decrease or increase in volume of the P2 substituent (Gly, Ser, or Leu) resulted in a decrease in this constant. Substrates with the highest keff values have Phe in the P3 position. As the hydrophobicity and volume of the amino acid in the P3 position decreased, the keff was reduced. The efficiency of substrates for hydrolysis by factor VIIa was enhanced by an increase of hydrophobicity in the P' structure. TF enhanced the amidolytic activity of the "family" of 38 substrates with ANSN in the P' position on an average of 58-fold.     

The role of "indirect" recognition in initiating rejection of skin grafts from major histocompatibility complex class II-deficient mice

In vitro studies have revealed several pathways by which T cells can respond to alloantigens, including CD4+ direct responses to allogeneic class II antigens, CD8+ direct responses to allogeneic class I antigens, and CD4+ "indirect" responses to peptides of alloantigens presented in association with responder class II molecules. In vivo studies of skin graft rejection, however, have so far provided clear evidence for the contribution of only the two direct pathways and not for indirect recognition. We have used major histocompatibility complex class II-deficient mice as donors to test the role of indirect recognition in rejection of skin grafts. Class II-deficient skin was always rejected without delay by normal recipients. Removal of recipient CD8+ cells (to leave the animals dependent on CD4+ function) or depletion of recipient CD4+ cells revealed that CD4+ cells were usually involved and sometimes absolutely required in this rapid rejection. Since the donor grafts lacked class II antigens, the CD4+ cells must have recognized donor antigens presented in association with recipient class II molecules. These results therefore indicate that indirect recognition can initiate rapid skin graft rejection.     

LINKING RIVER MANAGEMENT TO SPECIES CONSERVATION USING DYNAMIC LANDSCAPE‐SCALE MODELS

Efforts to conserve stream and river biota could benefit from tools that allow managers to evaluate landscape‐scale changes in species distributions in response to water management decisions. We present a framework and methods for integrating hydrology, geographic context and metapopulation processes to simulate effects of changes in streamflow on fish occupancy dynamics across a landscape of interconnected stream segments. We illustrate this approach using a 482 km² catchment in the southeastern US supporting 50 or more stream fish species. A spatially distributed, deterministic and physically based hydrologic model is used to simulate daily streamflow for sub‐basins composing the catchment. We use geographic data to characterize stream segments with respect to channel size, confinement, position and connectedness within the stream network. Simulated streamflow dynamics are then applied to model fish metapopulation dynamics in stream segments, using hypothesized effects of streamflow magnitude and variability on population processes, conditioned by channel characteristics. The resulting time series simulate spatially explicit, annual changes in species occurrences or assemblage metrics (e.g. species richness) across the catchment as outcomes of management scenarios. Sensitivity analyses using alternative, plausible links between streamflow components and metapopulation processes, or allowing for alternative modes of fish dispersal, demonstrate large effects of ecological uncertainty on model outcomes and highlight needed research and monitoring. Nonetheless, with uncertainties explicitly acknowledged, dynamic, landscape‐scale simulations may prove useful for quantitatively comparing river management alternatives with respect to species conservation. Published 2012. This article is a U.S. Government work and is in the public domain in the USA.     

Changes of bone-perfusion in osteosynthesis of the femur with a marrow-nail (author's transl)

The perfusion of the bone in the hind leg after osteosynthesis (nailing of the bone-marrow) was studied. In 11 shepherd dogs (bastards) an osteotomy of the femur was done; it was treated with a marrow-nail without boring the marrow-cavity. With the "tracer-microsphere"-method the perfusion of femur, tibia and talus of both hind legs was measured. Measurements were performed before and after surgery, in 10 dogs 2 weeks and in 8 dogs 6 weeks after surgery. Immediately after the operation the perfusion was reduced considerably in all the examined bones of the operated leg. Two weeks later the perfusion was increased in all bones of both hind limbs. In the cancellous bone of the femur the perfusion reached the original preoperative values after 6 weeks; in cortical bone a further increase of the perfusion was noted. This increase was most marked in the cortical bone of the operated femur; it was less in the cortical bone of the other bones.     

Recent Life Events Preceding Suicide Attempts in a Personality Disorder Sample: Findings From the Collaborative Longitudinal Personality Disorders Study.

Few studies have examined the relationship between life events, suicide attempts, and personality disorders (PDs), in spite of the strong associations between PDs and suicidal behavior, and the poor coping strategies often exhibited by these individuals. The authors examined whether participants with PDs who attempted suicide during the first 3 years of a prospective, longitudinal study were more likely to experience specific life events in the month during and preceding the suicide attempt. Of 489 participants with PDs, 61 attempted suicide during the 3-year, follow-up interval. Results indicated that negative life events, particularly those pertaining to love-marriage or crime-legal matters, were significant predictors of suicide attempts, even after controlling for baseline diagnoses of borderline PD, major depressive disorders, substance use disorders, and a history of childhood sexual abuse. Therefore, certain types of negative life events are unique risk factors for imminent suicide attempts among individuals with PDs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)