In yeast, a protein complex termed the ER-Mitochondria Encounter Structure (ERMES) tethers mitochondria to the endoplasmic reticulum. ERMES proteins are implicated in a variety of cellular functions including phospholipid synthesis, mitochondrial protein import, mitochondrial attachment to actin, polarized mitochondrial movement into daughter cells during division, and maintenance of mitochondrial DNA (mtDNA). The mitochondrial-anchored Gem1 GTPase has been proposed to regulate ERMES functions. Here, we show that ERMES and Gem1 have no direct role in the transport of phosphatidylserine (PS) from the ER to mitochondria during the synthesis of phosphatidylethanolamine (PE), as PS to PE conversion is not affected in ERMES or gem1 mutants. In addition, we report that mitochondrial inheritance defects in ERMES mutants are a secondary consequence of mitochondrial morphology defects, arguing against a primary role for ERMES in mitochondrial association with actin and mitochondrial movement. Finally, we show that ERMES complexes are long-lived, and do not depend on the presence of Gem1. Our findings suggest that the ERMES complex may have primarily a structural role in maintaining mitochondrial morphology.

SummaryThe intracellular protozoan Toxoplasma gondii lacks a de novo mechanism for cholesterol synthesis and therefore must scavenge this essential lipid from the host environment. In this study, we demonstrated that T. gondii diverts cholesterol from low-density lipoproteins for cholesteryl ester synthesis and storage in lipid bodies. We identified and characterized two isoforms of acyl-CoA:cholesterol acyltransferase

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Efficient transmission of Plasmodium species between humans and Anopheles mosquitoes is a major contributor to the global burden of malaria. Gametocytogenesis, the process by which parasites switch from asexual replication within human erythrocytes to produce male and female gametocytes, is a critical step in malaria transmission and Plasmodium genetic diversity. Nothing is known about the pathways that regulate gametocytogenesis and only few of the current drugs that inhibit asexual replication are also capable of inhibiting gametocyte development and blocking malaria transmission. Here we provide genetic and pharmacological evidence indicating that the pathway for synthesis of phosphatidylcholine in Plasmodium falciparum membranes from host serine is essential for parasite gametocytogenesis and malaria transmission. Parasites lacking the phosphoethanolamine N-methyltransferase enzyme, which catalyzes the limiting step in this pathway, are severely altered in gametocyte development, are incapable of producing mature-stage gametocytes, and are not transmitted to mosquitoes. Chemical screening identified 11 inhibitors of phosphoethanolamine N-methyltransferase that block parasite intraerythrocytic asexual replication and gametocyte differentiation in the low micromolar range. Kinetic studies in vitro as well as functional complementation assays and lipid metabolic analyses in vivo on the most promising inhibitor NSC-158011 further demonstrated the specificity of inhibition. These studies set the stage for further optimization of NSC-158011 for development of a class of dual activity antimalarials to block both intraerythrocytic asexual replication and gametocytogenesis.H uman malaria parasites exhibit a complex life cycle consisting of asexual phases within human hepatocytes and erythrocytes, with the latter directly responsible for disease manifestations. Within red blood cells, these parasites can also undergo gametocytogenesis, a process during which they interrupt their asexual replication and differentiate to form morphologically and functionally distinct sexual-stage gametocytes (1). These sexual forms serve as precursors for male and female gametes, which develop in the mosquito where they undergo mating, meiosis and several mitotic cycles to produce sporozoites. In Plasmodium falciparum, the causative agent of the most severe form of human malaria, the progression from immature stage I to mature stage V gametocytes takes ∼10 d (2). However, the biological processes that regulate gametocytogenesis remain unknown. Thorough understanding of these processes is crucial to the development of a new generation of dual activity antimalarials that can inhibit both infection and transmission.Phosphatidylcholine (PC), the predominant phospholipid produced by malaria parasites, plays essential structural and regulatory roles in parasite development and differentiation (reviewed in ref.3). Lipid metabolic and genetic studies in P. falciparum have demonstrated the presence of two pathways for PC biosynthesis (Fig. S1): ...

Mitochondrial membrane biogenesis requires the interorganelle transport of phospholipids. Phosphatidylserine (PtdSer) synthesized in the endoplasmic reticulum and related membranes (mitochondria-associated membrane (MAM)) is transported to the mitochondria by unknown gene products and decarboxylated to form phosphatidylethanolamine at the inner membrane by PtdSer decarboxylase 1 (Psd1p). We have designed a screen for strains defective in PtdSer transport (pstA mutants) between the endoplasmic reticulum and Psd1p that relies on isolating ethanolamine auxotrophs in suitable (psd2⌬) genetic backgrounds. Following chemical mutagenesis, we isolated an ethanolamine auxotroph that we designate pstA1-1. Using in vivo and in vitro phospholipid synthesis/transport measurements, we demonstrate that the pstA1-1 mutant is defective in PtdSer transport between the MAM and mitochondria. The gene that complements the growth defect and PtdSer transport defect of the pstA1-1 mutant is MET30, which encodes a substrate recognition subunit of the SCF (suppressor of kinetochore protein 1, cullin, F-box) ubiquitin ligase complex. Reconstitution of different permutations of MAM and mitochondria from wild type and pstA1-1 strains demonstrates that the MET30 gene product affects both organelles. These data provide compelling evidence that interorganelle PtdSer traffic is regulated by ubiquitination.

The compressive response of a NiTi shape-memory alloy is investigated at various strain rates using UCSD’s modified 12-in. Hopkinson pressure bar and a conventional Instron machine. To obtain a constant strain rate during the formation of a stress-induced martensite in a Hopkinson test, a copper tube of suitable dimensions is employed as a pulse shaper, since without a pulse shaper the strain rate of the sample varies significantly as its microstructure changes from austenite to martensite, whereas with proper pulse shaping techniques a nearly constant strain rate can be achieved over a certain deformation range. The NiTi shape-memory alloy shows a superelastic response for small strains at all considered strain rates and at room temperature, 296 K. At this temperature and below a certain strain rate, the stress–strain curves of the NiTi shape-memory alloy display two regimes: an elastic austenite regime and a transition (stress-induced martensite) regime. The transition stress of this material and the work-hardening rate in the stress-induced martensite regime increase with increasing strain rate, the latter reaching a steady state level and then rapidly increasing.

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Silicon photomultiplier (SiPM; also called a Geiger-mode avalanche photodiode) is a promising semiconductor photosensor in PET and PET/MRI because it is intrinsically MRI-compatible and has internal gain and timing properties comparable to those of a photomultiplier tube. In this study, we have developed a smallanimal PET system using SiPMs and lutetium gadolinium oxyorthosilicate (LGSO) crystals and performed physical evaluation and animal imaging studies to show the feasibility of this system. Methods: The SiPM PET system consists of 8 detectors, each of which comprises 2 · 6 SiPMs and 4 · 13 LGSO crystals. Each crystal has dimensions of 1.5 · 1.5 · 7 mm. The crystal face-toface diameter and axial field of view are 6.0 cm and 6.5 mm, respectively. Bias voltage is applied to each SiPM using a finely controlled voltage supply because the gain of the SiPM strongly depends on the supply voltage. The physical characteristics were studied by measuring energy resolution, sensitivity, and spatial resolution. Various mouse and rat images were obtained to study the feasibility of the SiPM PET system in in vivo animal studies. Reconstructed PET images using a maximum-likelihood expectation maximization algorithm were coregistered with animal CT images. Results: All individual LGSO crystals within the detectors were clearly distinguishable in flood images obtained by irradiating the detector using a 22 Na point source. The energy resolution for individual crystals was 25.8% 6 2.6% on average for 511-keV photopeaks. The spatial resolution measured with the 22 Na point source in a warm background was 1.0 mm (2 mm off-center) and 1.4 mm (16 mm off-center) when the maximum-likelihood expectation maximization algorithm was applied. A myocardial 18 F-FDG study in mice and a skeletal 18 F study in rats demonstrated the fine spatial resolution of the scanner. The feasibility of the SiPM PET system was also confirmed in the tumor images of mice using 18 F-FDG and 68 Ga-RGD and in the brain images of rats using 18 F-FDG. Conclusion: These results indicate that it is possible to develop a PET system using a promising semiconductor photosensor, which yielded reasonable PET performance in phantom and animal studies.