Cultivation, cryopreservation, and transcriptomic studies of host-adapted Cryptosporidium parvum and Cryptosporidium hominis using enteroids.

Cryptosporidium hominis and Cryptosporidium parvum are major causes of severe diarrhea. Comparative studies of them are hampered by the lack of effective cultivation and cryopreservation methods, especially for C. hominis. Here, we describe adapted murine enteroids for the cultivation and complete development of host-adapted C. parvum and C. hominis subtypes, producing oocysts infectious to mice. Using the system, we developed a cryopreservation method for Cryptosporidium isolates. In comparative RNA-seq analyses of C. hominis cultures, the enteroid system generated significantly more host and pathogen responses than the conventional HCT-8 cell system. In particular, the infection was shown to upregulate PI3K-Akt, Ras, TNF, NF-κB, IL-17, MAPK, and innate immunity signaling pathways and downregulate host cell metabolism, and had significantly higher expression of parasite genes involved in oocyst formation. Therefore, the enteroid system provides a valuable tool for comparative studies of the biology of divergent Cryptosporidium species and isolates.

Sulfonylurea receptor 2 (SUR2), intricate sensors for intracellular Mg-nucleotides.

SUR2, similar to SUR1, is a regulatory subunit of the ATP-sensitive potassium channel (KATP), which plays a key role in numerous important physiological processes and is implicated in various diseases. Recent structural studies have revealed that, like SUR1, SUR2 can undergo ligand-dependent dynamic conformational changes, transitioning between an inhibitory inward-facing conformation and an activating occluded conformation. In addition, SUR2 possesses a unique inhibitory Regulatory helix (R helix) that is absent in SUR1. The binding of the activating Mg-ADP to NBD2 of SUR2 competes with the inhibitory Mg-ATP, thereby promoting the release of the R helix and initiating the activation process. Moreover, the signal generated by Mg-ADP binding to NBD2 might be directly transmitted to the TMD of SUR2, prior to NBD dimerization. Furthermore, the C-terminal 42 residues (C42) of SUR2 might allosterically regulate the kinetics of Mg-nucleotide binding on NBD2. These distinctive properties render SUR2 intricate sensors for intracellular Mg-nucleotides.

The inhibition mechanism of the SUR2A-containing KATP channel by a regulatory helix.

KATP channels are metabolic sensors for intracellular ATP/ADP ratios, play essential roles in many physiological processes, and are implicated in a spectrum of pathological conditions. SUR2A-containing KATP channels differ from other subtypes in their sensitivity to Mg-ADP activation. However, the underlying structural mechanism remains poorly understood. Here we present a series of cryo-EM structures of SUR2A in the presence of different combinations of Mg-nucleotides and the allosteric inhibitor repaglinide. These structures uncover regulatory helix (R helix) on the NBD1-TMD2 linker, which wedges between NBD1 and NBD2. R helix stabilizes SUR2A in the NBD-separated conformation to inhibit channel activation. The competitive binding of Mg-ADP with Mg-ATP to NBD2 mobilizes the R helix to relieve such inhibition, allowing channel activation. The structures of SUR2B in similar conditions suggest that the C-terminal 42 residues of SUR2B enhance the structural dynamics of NBD2 and facilitate the dissociation of the R helix and the binding of Mg-ADP to NBD2, promoting NBD dimerization and subsequent channel activation.

Multiple introductions and recombination events underlie the emergence of a hyper-transmissible Cryptosporidium hominis subtype in the USA.

The parasite Cryptosporidium hominis is a leading cause of the diarrheal disease cryptosporidiosis, whose incidence in the United States has increased since 2005. Here, we show that the newly emerged and hyper-transmissible subtype IfA12G1R5 is now dominant in the United States. In a comparative analysis of 127 newly sequenced and 95 published C. hominis genomes, IfA12G1R5 isolates from the United States place into three of the 14 clusters (Pop6, Pop13, and Pop14), indicating that this subtype has multiple ancestral origins. Pop6 (IfA12G1R5a) has an East Africa origin and has recombined with autochthonous subtypes after its arrival. Pop13 (IfA12G1R5b) is imported from Europe, where it has recombined with the prevalent local subtype, whereas Pop14 (IfA12G1R5c) is a progeny of secondary recombination between Pop6 and Pop13. Selective sweeps in invasion-associated genes have accompanied the emergence of the dominant Pop14. These observations offer insights into the emergence and evolution of hyper-transmissible pathogens.

Controlling the emission linewidths of alloy quantum dots with asymmetric strain.

The emission linewidth of quantum dots (QDs) is one of the important optical properties, which is essential for the applications of QD lasers, high-quality displays, and biological imaging. However, we know less about controlling emission linewidth and its underlying mechanisms. Here we introduce a wurtzite ZnSe shell onto a wurtzite CdSe core to produce asymmetric strain due to their large, anisotropic lattice mismatch. Such asymmetric pressure induces significant splitting (ΔAB) between heavy-hole (hh) and light-hole (lh) in valence band (VB). We show that the emission intensity from the lh state (Elh) is significantly suppressed with the increasing ΔAB caused by the strong asymmetric strain. We demonstrate that the exciton-phonon coupling (EPC) is greatly inhibited under the anisotropic lattice strain. The alloying process between the core and shell occurs under the strong lattice strain and raises the longitudinal-optical (LO) phonon energy (ELO). Higher LO phonon energy declines LO phonon occupation numbers (NLO) and synergistically reduces the EPC. The asymmetrically strained alloy QDs ensemble exhibits highly bright emission with ultra-narrow linewidths of 13.8 nm (∼520 nm) and 15.8 nm (∼620 nm). This concept of band structure regulation via asymmetric strain can provide a new platform for high-quality QDs beyond the currently achieved.

A productive immunocompetent mouse model of cryptosporidiosis with long oocyst shedding duration for immunological studies.

OBJECTIVES: Studies on the pathogenesis and immune responses of Cryptosporidium infection and development of drugs and vaccines use mostly immunocompromised mouse models. In this study, we establish an immunocompetent mouse model of cryptosporidiosis with high intensity and long duration of infection. METHODS: We have obtained a Cryptosporidium tyzzeri isolate from laboratory mice, and infect adult C57BL/6 J mice experimentally with the isolate for determinations of infectivity, infection patterns, pathological changes, and transcriptomic responses. RESULTS: The isolate has an ID50 of 5.2 oocysts, with oocyst shedding lasting at high levels for >2 months. The oocyst shedding is boosted by immunosuppression of animals and suppressed by paromomycin treatment. The isolate induces strong inflammatory and acquired immune responses, but down-regulates the expression of α-defensins in epithelium. Comparative genomics analysis has revealed significant sequence differences from other isolates in subtelomeric genes. The down-regulation of the expression of α-defensins may be responsible for the high-intensity and long-lasting infection in this animal model. CONCLUSIONS: The immunocompetent mouse model of cryptosporidiosis developed has the advantages of high oocyst shedding intensity and long oocyst shedding duration. It provides an effective mechanism for the propagation of Cryptosporidium, evaluations of potential therapeutics, and studies of pathogen biology and immune responses.

Covalent Coupling-Stabilized Transition-Metal Sulfide/Carbon Nanotube Composites for Lithium/Sodium-Ion Batteries.

Transition-metal sulfides (TMSs) powered by conversion and/or alloying reactions are considered to be promising anode materials for advanced lithium-ion batteries (LIBs) and sodium-ion batteries (SIBs). However, the limited electronic conductivity and large volume expansion severely hinder their practical application. Herein, we report a covalent coupling strategy for TMS-based anode materials using amide linkages to bind TMSs and carbon nanotubes (CNTs). In the synthesis, the thiourea acts as not only the capping agent for morphology control but also the linking agent for the covalent coupling. As a proof of concept, the covalently coupled ZnS/CNT composite (CC-ZnS/CNT) has been prepared, with ZnS nanoparticles (∼10 nm) tightly anchored on CNT bundles. The compact ZnS-CNT heterojunctions are greatly beneficial to facilitating the electron/ion transfer and ensuring structural stability. Due to the strong coupling interaction between ZnS and CNTs, the composite presents prominent pseudocapacitive behavior and highly reversible electrochemical processes, thus leading to superior long-term stability and excellent rate capability, delivering reversible capacities of 333 mAh g-1 at 2 A g-1 over 4000 cycles for LIBs and 314 mAh g-1 at 5 A g-1 after 500 cycles for SIBs. Consequently, CC-ZnS/CNT exhibits great competence for applications in LIBs and SIBs, and the covalent coupling strategy is proposed as a promising approach for designing high-performance anode materials.

Mesoporous Graphitic Carbon-Encapsulated Fe2 O3 Nanocomposite as High-Rate Anode Material for Sodium-Ion Batteries.

A mesoporous graphitic carbon-encapsulated Fe2 O3 nanocomposite is synthesized as a superior anode material for sodium-ion batteries. A threefold strategy is adopted to achieve a high rate performance. First, the mesoporous structure with high specific surface area and large pore volume facilitates the transfer of electrolyte and accommodates the large volume change. Secondly, graphitic carbon encapsulation further improves the electronic conductivity of the nanocomposite. Finally, ultrafine Fe2 O3 nanocrystals effectively shorten the Na+ diffusion length. Consequently, this nanocomposite exhibits stable and fast Na+ storage, thus leading to excellent rate capability and cyclability. Pseudocapacitive behavior is found to dominate in the redox reactions, accounting for the outstanding rate and cycling performance. In addition, full cells, assembled with O3-Na0.9 [Cu0.22 Fe0.30 Mn0.48 ]O2 as cathodes, present good electrochemical performance.