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Liou, W.

Publications and source records attributed to Liou, W..

3 recordsLinked to original sources

Identification of a Putative Metal Transporter in the Apicoplast of Malaria Parasites

Plasmodium falciparum malaria parasites harbor an essential plastid organelle, called the apicoplast, which produces key metabolites required for organelle function and parasite viability. Apicoplast functions depend on iron and other metals, but the membrane transporters that mediate metal import into this organelle have been challenging to identify. Tetracycline antibiotics, including doxycycline, specifically target the apicoplast and can exhibit metal-dependent activity. Using tetracycline-affinity proteomics, we identified a doxycycline-interacting, uncharacterized transmembrane protein (UCT) targeted to the apicoplast periphery but not proteolytically processed. Although lacking sequence similarity to proteins of known function, UCT has a predicted structure with high similarity to pentameric CorA-family metal transporters that mediate metal uptake in other organisms. Functional tests revealed that UCT is dispensable for blood-stage asexual parasites, suggesting that the apicoplast has evolved redundant mechanisms for metal uptake. UCT knockdown in gametocytes, however, impairs the development of sexual parasites, which are critical for mosquito transmission. Our study identifies an apicoplast membrane protein with localization and structural properties that predict a role in metal transport into this key organelle. This discovery can provide a biochemical springboard to unravel broader apicoplast mechanisms of metal uptake across multiple stages of parasite development, including mosquito-stage parasites that display heightened UCT expression.

biochemistry↗

EVOH Hollow Fiber Encapsulation of Individual Caenorhabditis elegans for Transmission Electron Microscopy

We present a streamlined protocol for encapsulating individual Caenorhabditis elegans specimens for thin-section transmission electron microscopy (TEM) using ethylene vinyl alcohol (EVOH) copolymer hollow fibers. These transparent fibers, featuring a 175 m inner diameter, 25 m wall thickness, and an approximate molecular weight cutoff of 30 kDa, possess both hydrophilic and hydrophobic properties. This amphiphilic nature helps maintain a potent lumen structure even in dry, open air, enabling precise handling and orientation of small specimens prior to resin polymerization and eliminating the need for remounting. The protocol includes specimen transfer using wire loops, microwave-assisted chemical pre-fixation, dissection on a soft silicone substrate with micro-scissors, and manipulation of hollow fibers using insect pins. This approach preserves the identity of individual specimens throughout processing. During ultramicrotomy, the fiber serves as a visual guide, enhancing alignment and reproducibility of sectioning. As proof of principle, we demonstrate well-preserved ultrastructural features in chemically fixed, microwave- assisted resin-embedded C. elegans specimens. This method offers a robust and reproducible alternative to conventional gelatinous media encapsulation, particularly suited for TEM analysis of scarce or small biological samples.

cell biology↗

Long-term ketogenic diet causes hyperlipidemia, liver dysfunction, and glucose intolerance from impaired insulin trafficking and secretion in mice

A ketogenic diet (KD) is a very low-carbohydrate, very high-fat diet proposed to treat obesity and type 2 diabetes. While KD grows in popularity, its effects on metabolic health are understudied. Here we show that, in male and female mice, while KD protects against weight gain and induces weight loss, over long-term, mice develop hyperlipidemia, hepatic steatosis, and severe glucose intolerance. Unlike high fat diet-fed mice, KD mice are not insulin resistant and have low levels of insulin. Hyperglycemic clamp and ex vivo GSIS revealed cell-autonomous and whole-body impairments in insulin secretion. Major ER/Golgi stress and disrupted ER-Golgi protein trafficking was indicated by transcriptomic profiling of KD islets and confirmed by electron micrographs showing a dilated Golgi network likely responsible for impaired insulin granule trafficking and secretion. Overall, our results suggest long-term KD leads to multiple aberrations of metabolic parameters that caution its systematic use as a health promoting dietary intervention.

physiology↗