Plasma membrane association by N-acylation governs PKG function in Toxoplasma gondii

ABSTRACT Cyclic GMP (cGMP)-dependent protein kinase (protein kinase G [PKG]) is essential for microneme secretion, motility, invasion, and egress in apicomplexan parasites, However, the separate roles of two isoforms of the kinase that are expressed by some apicomplexans remain uncertain. Despite having identical regulatory and catalytic domains, PKG I is plasma membrane associated whereas PKG II is cytosolic in Toxoplasma gondii . To determine whether these isoforms are functionally distinct or redundant, we developed an auxin-inducible degron (AID) tagging system for conditional protein depletion in T. gondii . By combining AID regulation with genome editing strategies, we determined that PKG I is necessary and fully sufficient for PKG-dependent cellular processes. Conversely, PKG II is functionally insufficient and dispensable in the presence of PKG I . The difference in functionality mapped to the first 15 residues of PKG I , containing a myristoylated Gly residue at position 2 that is critical for membrane association and PKG function. Collectively, we have identified a novel requirement for cGMP signaling at the plasma membrane and developed a new system for examining essential proteins in T. gondii . IMPORTANCE Toxoplasma gondii is an obligate intracellular apicomplexan parasite and important clinical and veterinary pathogen that causes toxoplasmosis. Since apicomplexans can only propagate within host cells, efficient invasion is critically important for their life cycles. Previous studies using chemical genetics demonstrated that cyclic GMP signaling through protein kinase G (PKG)-controlled invasion by apicomplexan parasites. However, these studies did not resolve functional differences between two compartmentalized isoforms of the kinase. Here we developed a conditional protein regulation tool to interrogate PKG isoforms in T. gondii . We found that the cytosolic PKG isoform was largely insufficient and dispensable. In contrast, the plasma membrane-associated isoform was necessary and fully sufficient for PKG function. Our studies identify the plasma membrane as a key location for PKG activity and provide a broadly applicable system for examining essential proteins in T. gondii . </jats:p

Methadone and Corrected QT Prolongation in Pain and Palliative Care Patients: A Case–Control Study

Background: Methadone (ME) is commonly used in pain and palliative care (PPC) patients with refractory pain or intolerable opioid adverse effects (AEs). A unique ME AE is its corrected QT (QTc) interval prolongation risk, but most evidence exists in methadone maintenance therapy patients. Objective: Our goal was to identify QTc interval prolongation risk factors in PPC patients receiving ME and other medications known to prolong the QTc interval and develop a risk stratification tool. Design: We performed a case–control study of adult inpatients receiving ME for pain management. Settings/Subjects: Adult inpatients receiving ME with a QTc \u3e470 msec (males) and \u3e480 msec (females) were matched 1:2 according to age, history of QTc prolongation, and gender with ME patients who did not have a prolonged QTc interval. QTc prolongation risk factors were collected for both groups. Covariates were analyzed using conditional logistic regression. Classification and regression tree analysis was used to identify the ME dose associated with QTc prolongation. Results: Predictors of QTc prolongation included congestive heart failure (CHF) (OR: 11.9; 95% CI: 3.7–38.2; p \u3c 0.00), peptic ulcer disease (PUD) (odds ratio [OR]: 8.3; 95% confidence interval [95% CI]: 2.4–28.9; p \u3c 0.00), hypokalemia (OR: 6.5; 95% CI: 1.5–28.2; p \u3c 0.01), rheumatologic diseases (OR: 4.7; 95% CI: 1.6–13.9; p \u3c 0.00), taking medications with a known torsades de pointes (TdP) risk (OR: 4.4; 95% CI: 1.8–10.7; p \u3c 0.01), malignancy (OR: 3.3; 95% CI: 1.2–9.3; p \u3c 0.03), hypocalcemia (OR: 2.1; 95% CI: 0.9–4.8; p \u3c 0.07), and ME doses \u3e45 mg per day (OR: 1.9; 95% CI: 0.8–4.8; p \u3c 0.16). Mild liver disease was protective against QTc prolongation (OR: 0.05; 95% CI: 0.0–0.46; p \u3c 0.01). Conclusions: Predictors of QTc prolongation in our multivariate conditional logistic regression model included CHF, PUD, hypokalemia, rheumatologic disorders, use of medications with a known TdP risk, malignancy, hypocalcemia, and ME doses \u3e45 mg per day

Catalyst for carbon monoxide oxidation

A catalyst is disclosed for the combination of CO and O2 to form CO2, which includes a platinum group metal (e.g., platinum); a reducable metal oxide having multiple valence states (e.g., SnO2); and a compound which can bind water to its structure (e.g., silica gel). This catalyst is ideally suited for application to high-powered pulsed, CO2 lasers operating in a sealed or closed-cycle condition

Rural Land-Use Trends in the Conterminous United States, 1950-2000.

In order to understand the magnitude, direction, and geographic distribution of land-use changes, we evaluated land-use trends in U.S. counties during the latter half of the 20th century. Our paper synthesizes the dominant spatial and temporal trends in population, agriculture, and urbanized land uses, using a variety of data sources and an ecoregion classification as a frame of reference. A combination of increasing attractiveness of nonmetropolitan areas in the period 1970–2000, decreasing household size, and decreasing density of settlement has resulted in important trends in the patterns of developed land. By 2000, the area of low-density, exurban development beyond the urban fringe occupied nearly 15 times the area of higher density urbanized development. Efficiency gains, mechanization, and agglomeration of agricultural concerns has resulted in data that show cropland area to be stable throughout the Corn Belt and parts of the West between 1950 and 2000, but decreasing by about 22% east of the Mississippi River. We use a regional case study of the Mid-Atlantic and Southeastern regions to focus in more detail on the land-cover changes resulting from these dynamics. Dominating were land-cover changes associated with the timber practices in the forested plains ecoregions and urbanization in the piedmont ecoregions. Appalachian ecoregions show the slowest rates of landcover change. The dominant trends of tremendous exurban growth, throughout the United States, and conversion and abandonment of agricultural lands, especially in the eastern United States, have important implications because they affect large areas of the country, the functioning of ecological systems, and the potential for restoratio