Cell wall chitosan is necessary for virulence in the opportunistic pathogen Cryptococcus neoformans

Cryptococcus neoformans is an opportunistic fungal pathogen that causes meningoencephalitis. Its cell wall is composed of glucans, proteins, chitin, and chitosan. Multiple genetic approaches have defined a chitosan-deficient syndrome that includes slow growth and decreased cell integrity. Here we demonstrate chitosan is necessary for virulence and persistence in the mammalian host

Soybean Yield, Evapotranspiration, Water Productivity, And Soil Water Extraction Response To Subsurface Drip Irrigation And Fertigation

Soybean [Glycine max (L.) Merr.] yield, irrigation water use efficiency (IWUE), crop water use efficiency (CWUE), evapotranspiration water use efficiency (ETWUE), and soil water extraction response to eleven treatments of full, limited, or delayed irrigation versus a rainfed control were investigated using a subsurface drip irrigation (SDI) system at a research site in south-central Nebraska. The SDI system laterals were 0.40 m deep in every other row middle of 0.76 m spaced plant rows. Actual evapotranspiration (ETa) was quantified in all treatments and used to schedule irrigation events on a 100% ETa replacement basis in all but three of the eleven treatments (i.e., 75% ETa replacement was used in two, and 60% ETa replacement was used in one). The irrigation amount (Ia) applied at each event was 100% of the ETa amount, except for two 100% ETa treatments in which only 65% or 50% of the water needed to cover the treatment plot area was applied to enable a test of a partial surface area-based irrigation approach. The first irrigation event was delayed until soybean stage R3 (begin pod) in two 100% Ia treatments, but thereafter they were irrigated with either 100% or 75% ETa replacement. Two 100% ETa and 100% Ia treatments also were used to evaluate soybean response to nitrogen (N) application methods (i.e., a preplant method versus N injection using the SDI system). Soybean ETa varied from 452 mm for the rainfed treatment to 600 mm (30% greater) for the fully irrigated treatment (100% ETa and 100% Ia) in 2007, and from 473 to 579 mm (20% greater) for the same treatments, respectively, in 2008. Among the irrigated treatments, 100% ETa and 65% Ia had the lowest 2007 ETa value (557 mm), whereas 100% ETa and 50% Ia had the lowest 2008 ETa (498 mm). The 100%, 75%, and 60% ETa treatments with 100% Ia had respective actual ETa values that declined linearly in 2008 (i.e., 579, 538, and 498 mm), but not in 2007. Seasonal totals for ETa versus Ia exhibited a linear relationship (R2 = 0.68 in 2007 and R2 = 0.67 in 2008). Irrigation enhanced soybean yields from rainfed yield baselines of 4.04 ton ha-1 in 2007 and 4.82 ton ha-1 in 2008) to a maximum of 4.94 ton ha-1 attained in 2007 with the delay to R3 irrigation treatment (its yield was significantly greater, p \u3c 0.05, than that of the seven other treatments) and 4.97 ton ha-1 attained in 2008 with the 100% ETa and 100% Ia preplant N treatment. Seed yield had a quadratic relationship with irrigation water applied and a linear relationship with ETa that was stronger in the drier year of 2007. Each 25.4 mm incremental increase in seasonal irrigation water applied increased soybean yield by 0.323 ton ha-1 (beyond the intercept) in 2007 and by 0.037 ton ha-1 in 2008. Each 25.4 mm increase in ETa generated a yield increase of 0.114 ton ha-1 (beyond the intercept) in 2007, but only 0.02 ton ha-1 in the wetter year of 2008. This research demonstrated that delaying the onset of irrigation until the R3 stage and practicing full irrigation thereafter for soybean grown on silt loam soils resulted in yields (and crop water productivity) that were similar to full-season irrigation scheduling strategies, and this result may be applicable in other regions with edaphic and climatic characteristics similar to those in south-central Nebraska

Soybean Yield, Evapotranspiration, Water Productivity, And Soil Water Extraction Response To Subsurface Drip Irrigation And Fertigation

Soybean [Glycine max (L.) Merr.] yield, irrigation water use efficiency (IWUE), crop water use efficiency (CWUE), evapotranspiration water use efficiency (ETWUE), and soil water extraction response to eleven treatments of full, limited, or delayed irrigation versus a rainfed control were investigated using a subsurface drip irrigation (SDI) system at a research site in south-central Nebraska. The SDI system laterals were 0.40 m deep in every other row middle of 0.76 m spaced plant rows. Actual evapotranspiration (ETa) was quantified in all treatments and used to schedule irrigation events on a 100% ETa replacement basis in all but three of the eleven treatments (i.e., 75% ETa replacement was used in two, and 60% ETa replacement was used in one). The irrigation amount (Ia) applied at each event was 100% of the ETa amount, except for two 100% ETa treatments in which only 65% or 50% of the water needed to cover the treatment plot area was applied to enable a test of a partial surface area-based irrigation approach. The first irrigation event was delayed until soybean stage R3 (begin pod) in two 100% Ia treatments, but thereafter they were irrigated with either 100% or 75% ETa replacement. Two 100% ETa and 100% Ia treatments also were used to evaluate soybean response to nitrogen (N) application methods (i.e., a preplant method versus N injection using the SDI system). Soybean ETa varied from 452 mm for the rainfed treatment to 600 mm (30% greater) for the fully irrigated treatment (100% ETa and 100% Ia) in 2007, and from 473 to 579 mm (20% greater) for the same treatments, respectively, in 2008. Among the irrigated treatments, 100% ETa and 65% Ia had the lowest 2007 ETa value (557 mm), whereas 100% ETa and 50% Ia had the lowest 2008 ETa (498 mm). The 100%, 75%, and 60% ETa treatments with 100% Ia had respective actual ETa values that declined linearly in 2008 (i.e., 579, 538, and 498 mm), but not in 2007. Seasonal totals for ETa versus Ia exhibited a linear relationship (R2 = 0.68 in 2007 and R2 = 0.67 in 2008). Irrigation enhanced soybean yields from rainfed yield baselines of 4.04 ton ha-1 in 2007 and 4.82 ton ha-1 in 2008) to a maximum of 4.94 ton ha-1 attained in 2007 with the delay to R3 irrigation treatment (its yield was significantly greater, p \u3c 0.05, than that of the seven other treatments) and 4.97 ton ha-1 attained in 2008 with the 100% ETa and 100% Ia preplant N treatment. Seed yield had a quadratic relationship with irrigation water applied and a linear relationship with ETa that was stronger in the drier year of 2007. Each 25.4 mm incremental increase in seasonal irrigation water applied increased soybean yield by 0.323 ton ha-1 (beyond the intercept) in 2007 and by 0.037 ton ha-1 in 2008. Each 25.4 mm increase in ETa generated a yield increase of 0.114 ton ha-1 (beyond the intercept) in 2007, but only 0.02 ton ha-1 in the wetter year of 2008. This research demonstrated that delaying the onset of irrigation until the R3 stage and practicing full irrigation thereafter for soybean grown on silt loam soils resulted in yields (and crop water productivity) that were similar to full-season irrigation scheduling strategies, and this result may be applicable in other regions with edaphic and climatic characteristics similar to those in south-central Nebraska

The Aminoalkylindole BML-190 Negatively Regulates Chitosan Synthesis via the Cyclic AMP/Protein Kinase A1 Pathway in Cryptococcus neoformans

Cryptococcus neoformans can cause fatal meningoencephalitis in patients with AIDS or other immunocompromising conditions. Current antifungals are suboptimal to treat this disease; therefore, novel targets and new therapies are needed. Previously, we have shown that chitosan is a critical component of the cryptococcal cell wall and is required for survival in the mammalian host and that chitosan deficiency results in rapid clearance from the mammalian host. We had also identified several specific proteins that were required for chitosan biosynthesis, and we hypothesize that screening for compounds that inhibit chitosan biosynthesis would identify additional genes/proteins that influence chitosan biosynthesis. To identify these compounds, we developed a robust and novel cell-based flow cytometry screening method to identify small-molecule inhibitors of chitosan production. We screened the ICCB Known Bioactives library and identified 8 compounds that reduced chitosan in C. neoformans We used flow cytometry-based counterscreens and confirmatory screens, followed by a biochemical secondary screen to refine our primary screening hits to 2 confirmed hits. One of the confirmed hits that reduced chitosan content was the aminoalkylindole BML-190, a known inverse agonist of mammalian cannabinoid receptors. We demonstrated that BML-190 likely targets the C. neoformans G-protein-coupled receptor Gpr4 and, via the cyclic AMP (cAMP)/protein kinase A (PKA) signaling pathway, contributes to an intracellular accumulation of cAMP that results in decreased chitosan. Our discovery suggests that this approach could be used to identify additional compounds and pathways that reduce chitosan biosynthesis and could lead to potential novel therapeutics against C. neoformans IMPORTANCE Cryptococcus neoformans is a fungal pathogen that kills approximately 200,000 people every year. The cell wall is an essential organelle that protects fungi from the environment. Chitosan, the deacetylated form of chitin, has been shown to be an essential component of the cryptococcal cell wall during infection of a mammalian host. In this study, we screened a set of 480 compounds, which are known to have defined biological activities, for activity that reduced chitosan production in C. neoformans Two of these compounds were confirmed using an alternative method of measuring chitosan, and one of these was demonstrated to impact the cAMP signal transduction pathway. This work demonstrates that the cAMP pathway regulates chitosan biosynthesis in C. neoformans and validates that this screening approach could be used to find potential antifungal agents

Congenital Prosopagnosia: Multistage Anatomical and Functional Deficits in Face Processing Circuitry

Face recognition is a primary social skill which depends on a distributed neural network. A pronounced face recognition deficit in the absence of any lesion is seen in congenital prosopagnosia. This study investigating 24 congenital prosopagnosic subjects and 25 control subjects aims at elucidating its neural basis with fMRI and voxel-based morphometry. We found a comprehensive behavioral pattern, an impairment in visual recognition for faces and buildings that spared long-term memory for faces with negative valence. Anatomical analysis revealed diminished gray matter density in the bilateral lingual gyrus, the right middle temporal gyrus, and the dorsolateral prefrontal cortex. In most of these areas, gray matter density correlated with memory success. Decreased functional activation was found in the left fusiform gyrus, a crucial area for face processing, and in the dorsolateral prefrontal cortex, whereas activation of the medial prefrontal cortex was enhanced. Hence, our data lend strength to the hypothesis that congenital prosopagnosia is explained by network dysfunction and suggest that anatomic curtailing of visual processing in the lingual gyrus plays a substantial role. The dysfunctional circuitry further encompasses the fusiform gyrus and the dorsolateral prefrontal cortex, which may contribute to their difficulties in long-term memory for complex visual information. Despite their deficits in face identity recognition, processing of emotion related information is preserved and possibly mediated by the medial prefrontal cortex. Congenital prosopagnosia may, therefore, be a blueprint of differential curtailing in networks of visual cognition

Cross talk between the Cell Wall Integrity and Cyclic AMP/Protein Kinase A pathways in Cryptococcus neoformans

Cryptococcus neoformans is a fungal pathogen of immunocompromised people that causes fatal meningitis. The fungal cell wall is essential to viability and pathogenesis of C. neoformans, and biosynthesis and repair of the wall is primarily controlled by the cell wall integrity (CWI) signaling pathway. Previous work has shown that deletion of genes encoding the four major kinases in the CWI signaling pathway, namely, PKC1, BCK1, MKK2, and MPK1 results in severe cell wall phenotypes, sensitivity to a variety of cell wall stressors, and for Mpk1, reduced virulence in a mouse model. Here, we examined the global transcriptional responses to gene deletions of BCK1, MKK2, and MPK1 compared to wild-type cells. We found that over 1,000 genes were differentially expressed in one or more of the deletion strains, with 115 genes differentially expressed in all three strains, many of which have been identified as genes regulated by the cyclic AMP (cAMP)/protein kinase A (PKA) pathway. Biochemical measurements of cAMP levels in the kinase deletion strains revealed significantly less cAMP in all of the deletion strains compared to the wild-type strain. The deletion strains also produced significantly smaller capsules than the wild-type KN99 strain did under capsule-inducing conditions, although the levels of capsule they shed were similar to those shed by the wild type. Finally, addition of exogenous cAMP led to reduced sensitivity to cell wall stress and restored surface capsule to levels near those of wild type. Thus, we have direct evidence of cross talk between the CWI and cAMP/PKA pathways that may have important implications for regulation of cell wall and capsule homeostasis

Magnetic field effect on the dielectric constant of glasses: Evidence of disorder within tunneling barriers

The magnetic field dependence of the low frequency dielectric constant $e_r$(H) of a structural glass a - SiO2 + xCyHz was studied from 400 mK to 50 mK and for H up to 3T. Measurement of both the real and the imaginary parts of $e_r$ is used to eliminate the difficult question of keeping constant the temperature of the sample while increasing H: a non-zero $e_r$(H) dependence is reported in the same range as that one very recently reported on multicomponent glasses. In addition to the recently proposed explanation based on interactions, the reported $e_r$(H) is interpreted quantitatively as a consequence of the disorder lying within the nanometric barriers of the elementary tunneling systems of the glass.Comment: latex Bcorrige1.tex, 5 files, 4 figures, 7 pages [SPEC-S02/009

Shaping the Phase of a Single Photon

While the phase of a coherent light field can be precisely known, the phase of the individual photons that create this field, considered individually, cannot. Phase changes within single-photon wave packets, however, have observable effects. In fact, actively controlling the phase of individual photons has been identified as a powerful resource for quantum communication protocols. Here we demonstrate the arbitrary phase control of a single photon. The phase modulation is applied without affecting the photon's amplitude profile and is verified via a two-photon quantum interference measurement, which can result in the fermionic spatial behaviour of photon pairs. Combined with previously demonstrated control of a single photon's amplitude, frequency, and polarisation, the fully deterministic phase shaping presented here allows for the complete control of single-photon wave packets.Comment: 4 pages, 4 figure