Isolation and Characterization of Strains Defective in Vacuolar Ornithine Permease inNeurospora crassa

Vacuolar uptake of ornithine and lysine was characterized inNeurospora crassausing a cupric ion permeabilization system. Michaelis constants were measured as 1.4 mM for lysine and 11.0 mM for ornithine, and maximal velocities were determined. Vacuolar lysine uptake was shown to be inhibited competitively byl-arginine and histidine while ornithine uptake was inhibited by a variety of amino acids. Strains defective in the vacuolar ornithine permease were isolated using a filtration enrichment method. Two isolates—RSC-39 and RSC-63—had a reduced ability to accumulate ornithine. Vacuolar uptake of amino acids was measured using cupric ion-permeabilized mycelia; both strains had reduced ornithine uptake while lysine uptake and arginine uptake were normal. For both isolates, both the Michaelis constant and the maximal velocity for ornithine uptake were reduced compared to those of wild type. These results suggest that both strains are defective in the gene which encodes the vacuolar ornithine permease.     

Kinetic parameters of nitrate uptake by different catch crop species: effects of low temperatures or previous nitrate starvation

The pollution of aquifers by NO^?₃ in temperate environments is aggravated by farming practices that leave the ground bare during winter. The use of catch crops during this time may decrease nitrate loss from the soil. Nitrate uptake by several catch crop species (Brassica napus L., Sinapis alba L., Brassica rapa L., Raphanus sativus L., Trifolium alexandrinum L., Trifolium incarnatum L., Phacelia tanacetifolia Benth., Lolium perenne L., Lolium multiflorum Lam. and Secale cereale L.) was here studied in relation to transpiration rate and low temperatures applied to the whole plant or to roots only. The Michaelis constant (K_m), maximum uptake rate (V_max), time of induction and contributions of inducible and constitutive mechanisms were estimated from measurements of NO^?₃ depletion in the uptake medium. There were large differences between species, with K_m (μM) values ranging between 5.12 ± 0.64 (Trifolium incarnatum) and 36.4 ± 1.97 (Lolium perenne). Maximum NO^?₃ uptake rates expressed per unit root weight were influenced by ageing, temperature and previous NO^?₃ nutrition. They were also closely correlated with water flow through the roots and with shoot/root ratio of these species. The combined results from all species and treatments showed that V_max increased with shoot/root ratio, suggesting a regulatory role for the shoots in NO^?₃ uptake. Overall, the results showed a great diversity in NO^?₃ uptake characteristics between species in terms of kinetic parameters, contribution of the constitutive system (100% of total uptake in ryegrass, nil in Fabaceae) and time of induction.     

Dissecting the signaling pathways involved in the function of sperm flagellum

The mammalian flagellum is a specific type of motile cilium required for sperm motility and male fertility. Effective flagellar movement is dependent on axonemal function, which in turn relies on proper ion homeostasis within the flagellar compartment. This ion homeostasis is maintained by the concerted function of ion channels and transporters that initiate signal transduction pathways resulting in motility changes. Advances in electrophysiology and super-resolution microscopy have helped to identify and characterize new regulatory modalities of the mammalian flagellum. Here, we discuss what is currently known about the regulation of flagellar ion channels and transporters that maintain sodium, potassium, calcium, and proton homeostasis. Identification of new regulatory elements and their specific roles in sperm motility is imperative for improving diagnostics of male infertility.     

Isomeric lipid signatures reveal compartmentalized fatty acid metabolism in cancer

The cellular energy and biomass demands of cancer drive a complex dynamic between uptake of extracellular FAs and their de novo synthesis. Given that oxidation of de novo synthesized FAs for energy would result in net-energy loss, there is an implication that FAs from these two sources must have distinct metabolic fates; however, hitherto, all FAs have been considered part of a common pool. To probe potential metabolic partitioning of cellular FAs, cancer cells were supplemented with stable isotope-labeled FAs. Structural analysis of the resulting glycerophospholipids revealed that labeled FAs from uptake were largely incorporated to canonical (sn-) positions on the glycerol backbone. Surprisingly, labeled FA uptake also disrupted canonical isomer patterns of the unlabeled lipidome and induced repartitioning of n-3 and n-6 PUFAs into glycerophospholipid classes. These structural changes support the existence of differences in the metabolic fates of FAs derived from uptake or de novo sources and demonstrate unique signaling and remodeling behaviors usually hidden from conventional lipidomics.     

Physical and Microbiological Analysis of Sandstone Deterioration in the Argentine Jesuit Missions

Mainly cyanobacteria, but also algae and fungi, were identified on the Jesuit missions San Ignacio Miní, Loreto, Santa María and Santa Ana, in NE Argentina. Many organisms were deeply pigmented, indicative of stressful conditions. San Ignacio samples of itacurú rock showed higher colonization and biodiversity, related to higher porosity allowing entry of water and nutrients. Sandstone from other locations was arenitous, containing 95% silicates. Physicochemical analyses suggested mobilization of ions, particularly Fe, from within the stone to the surface. This intense deteriorating colonization, together with climatic weathering, transforms the sandstone to a “protosoil,” for growth of bryophytes and higher plants.     

The transepithelial shunt pathway in the renal proximal tubule of newt kidney

The transepithelial shunt pathway of newt proximal tubule was examined with glass micro-electrode and electron microscopic methods. The input resistance of the peritubular (basal) membrane and tubular wall were found to be $\text{4.2 ± 0.1 · 10}{}^6$ ($\text{mean ± S.E.}\text{, n = 16}$) and $\text{11.4 ± 0.2 · 10}{}^4\text{(n = 11)}$, respectively. The input resistance of the peritubular membrane was approximately 40-times larger than that of the tubular wall. When the kidneys were perfused in a lanthanum solution, the lanthanum ions were then observed in the junctional complexes and in the intercellular spaces on both the basal and apical sides. The results indicate that the electrical shunt pathway corresponds to the apical junctional complexes and the intercellular spaces, and that the tight junctions are not truly ‘tight’ for the transepithelial movement of small ions in the proximal tubule of the newt kidney.     

Root development in winter wheat grown at different N/P supply: Root length patterns and N-P interactions in phosphate uptake

Effects of different N/P ratios on several root parameters and on net P uptake were studied in winter wheat, Triticum aestivum cv. Starke II, grown in water culture. In the First experiment N/P ratios of (0/4, 2/3, 4/2, 6/1 and 8/0) were used, and plants were harvested at age 3, 5, 8, 11 and 14 days. In the second experiment N/P ratios of 6/1, 10/1, 15/1, 17/1, 20/1 and 25/1 were applied at two different N,P levels. Root length and number were determined using a digitizer connected to a computer. In the first experiment. the 6/1 N/P ratio gave the largest plants at day 14, and growth decreased with decreasing N/P ratio, The same pattern was found fur lateral root length and root number (seminal and lateral). In the second experiment the root weights decreased with increasing N/P ratio within each level. Lateral root number and overall length decreased with increasing N/P ratio at both levels as did the average lateral root length at the high N,P level. At the low N.P level, average lateral root length was about the same at all N/P ratios. Increasing the N/P ratio increased net uptake of P at the low N,P level, but decreased net P uptake at the high N,P level. Net P uptake increased with increasing P concentration in the roots and then decreased with further increase in P concentration. Net P uptake based on calculated root length [m (g root)⁻¹] showed no significant deviation from weight-based uptake plots. The effect of N and P on root structure is discussed as well as the interaction of N and P in P uptake. The relevance of a proper basis for expressing root activity is stressed.     

K+(Rb+) and Ca2+ fluxes in young winter wheat plants exposed to sub-zero temperatures

Ice crystal formation temperature was determined in the region of the crown in one group of 7-day-old intact unhardened high-salt plants of winter wheat (Triticum aestivum L. cv. Weibulls Starke II) with TA (Thermal Analysis) and DTA (Differential Thermal Analysis) methods. After exposure of another group of plants, grown for the first 7 days in the same way as the first group, to various sub-zero temperatures (-1 to 5°C), influx in roots of Rb⁺(⁸⁶Rb⁺) and Ca²⁺(⁴⁵Ca²⁺) and contents of K⁺ and Ca²⁺ were determined at intervals during 7 days of recovery. Ice crystal formation in the crown tissue was probably extracellular and took place at about -4°C. There was a large loss of K⁺ from the roots after treatment at sub-zero temperatures. This loss increased as the temperature of the sub-zero treatment decreased. During recovery, roots of plants exposed to -1, -2 and -3°C gradually reabsorbed K⁺. Reabsorption of K⁺ in roots of plants exposed to -4°C was greatly impaired. Rb⁺ influx decreased and Ca²⁺ influx increased after sub-zero temperature treatments of the plants. Active Rb⁺ influx mechanisms and active extrusion of Ca²⁺ were impaired or irreversibly damaged by the exposure. While Rb⁺ influx mechanisms were apparently repaired during recovery in plants exposed to temperatures down to -3°C, Ca²⁺ extrusion mechanisms were not. The temperature for ice crystal formation in the region of the crown tissue coincides with the temperature at which the plants lost the ability to reabsorb K⁺ and to repair Rb⁺ influx mechanisms during the recovery period. Plants were lethally damaged at temperatures below ?4°C.