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Kinetoplastids are infamous parasites that include trypanosomes and Leishmania species. Here, we developed an anti-Leishmania nano-drug using ultra-small functional maghemite (γ-Fe2O3) nanoparticles (NPs) that were surface-doped by [CeLn]3/4+ to enable effective binding of the polycationic polyethylenebyimine (PEI) polymer by coordinative chemistry. This resulting nano-drug is cytolytic in-vitro to both Trypanosoma brucei parasites, the causative agent of sleeping sickness, as well as to three Leishmania species. The nano-drug induces the rupture of the single lysosome present in these parasites attributed to the PEI, leading to cytolysis. To evaluate the efficacy of a “cream-based” version of the nano-drug, which was termed “Nano-Leish-IL” for topical treatment of cutaneous leishmaniasis (CL), we developed a rapid screening method utilizing T. brucei parasites involved in social motility and demonstrated that functional NPs arrested the migration of the parasites. This assay presents a surrogate system to rapidly examine the efficacy of “cream-based” drugs in topical preparations against leishmaniasis, and possibly other dermal infectious diseases. The resulting Nano-Leish-IL topical preparation eliminated L. major infection in mice. Thus, this study presents a novel efficient nano-drug targeting the single lysosome of kinetoplastid parasites.

These are some data derived from the MAGE model of the thermosphere ionosphere. Neutral density and GOLD equivalent temperatures are given here. More descriptions are as below: dimensions: nlats_gold = 52 ; nlons_gold = 46 ; ndays = 2 ; nut_gold = 4 ; n_height_mage = 92 ; variables: float latitude_gold(nlats_gold, nlons_gold) ; latitude_gold:units = "degree_north; -90 to +90" ; float longitude_gold(nlats_gold, nlons_gold) ; longitude_gold:units = "degree_east; -180 to +180" ; float height_MAGE(n_height_mage) ; height_MAGE:units = "km" ; height_MAGE:long_name = "MAGE geometrical heights" ; string dates(ndays) ; dates:long_name = "YYYYMMDD format date; e.g., 20181130 for November 30, 2018" ; byte ut_times(nut_gold) ; ut_times:units = "Universal Time (in hrs) when GOLD measurements were made" ; float MAGE_Teff(nut_gold, nlats_gold, nlons_gold) ; MAGE_Teff:units = "Kelvin" ; MAGE_Teff:long_name = "GOLD equivalent temperature (Teff) from MAGE" ; float MAGE_density(ndays, n_height_mage) ; MAGE_density:units = "gm/cc" ; MAGE_density:long_name = "MAGE Neutral density; during 10 to 19 UT and 60S to 60N " ; // global attributes: :description = "GOLD Equivalent Temperature and from MAGE model and neutral density; missing data: NaN" ; :history = "Created 2022-07-06 23:26:41.538906"

In the parasite Trypanosoma brucei, the causative agent of human African sleeping sickness, all mRNAs are trans-spliced to generate a common 5′ exon derived from the spliced leader (SL) RNA. Perturbations of protein translocation across the endoplasmic reticulum (ER) induce the spliced leader RNA silencing (SLS) pathway. SLS activation is mediated by a serine-threonine kinase, PK3, which translocates from the cytosolic face of the ER to the nucleus, where it phosphorylates the TATA-binding protein TRF4, leading to the shutoff of SL RNA transcription, followed by induction of programmed cell death. Here, we demonstrate that SLS is also induced by depletion of the essential ER-resident chaperones BiP and calreticulin, ER oxidoreductin 1 (ERO1), and the Golgi complex-localized quiescin sulfhydryl oxidase (QSOX). Most strikingly, silencing of Rhomboid-like 1 (TIMRHOM1), involved in mitochondrial protein import, also induces SLS. The PK3 kinase, which integrates SLS signals, is modified by phosphorylation on multiple sites. To determine which of the phosphorylation events activate PK3, several individual mutations or their combination were generated. These mutations failed to completely eliminate the phosphorylation or translocation of the kinase to the nucleus. The structures of PK3 kinase and its ATP binding domain were therefore modeled. A conserved phenylalanine at position 771 was proposed to interact with ATP, and the PK3F771L mutation completely eliminated phosphorylation under SLS, suggesting that the activation involves most if not all of the phosphorylation sites. The study suggests that the SLS occurs broadly in response to failures in protein sorting, folding, or modification across multiple compartments. IMPORTANCE In this study, we found that SLS is induced by depletion of the essential ER-resident chaperones BiP and calreticulin, ER oxidoreductin 1 (ERO1), and the Golgi complex-localized quiescin sulfhydryl oxidase (QSOX). Most strikingly, silencing of Rhomboid-like 1 (TIMRHOM1), involved in mitochondrial protein import, also induces SLS. We also report on the autophosphorylation of PK3 during SLS induction. This study has implications for our understanding of how trypanosomes keep the homeostasis between the ER and the mitochondria and suggests that PK3 may participate in the connection between these two organelles. The pathway, when induced, leads to the suicide of these parasites, and its induction offers a potential novel drug target against these parasites.

We have observed electron impact fluorescence from CO2 to excite the Cameron bands (CBs), CO (a 3Π → X 1Σ+; 180–280 nm), the first-negative group (1NG) bands, CO+ (B 2Σ+ → X 2Σ+; 180–320 nm), the fourth-positive group (4PG) bands, CO (A 1Π → X 1Σ+; 111–280 nm), and the UV doublet, CO2 + ( B ˜ 2 Σ u + → X ˜ 2 Π g ; 288.3 and 289.6 nm) in the ultraviolet (UV). This wavelength range matches the spectral region of past and present spacecraft equipped to observe UV dayglow and aurora emissions from the thermospheres (100–300 km) of Mars and Venus. Our large vacuum system apparatus is able to measure the emission cross sections of the strongest optically forbidden UV transitions found in planetary spectra. Based on our cross-sectional measurements, previous CB emission cross-sectional errors exceed a factor of 3. The UV doublet lifetime is perturbed through B ˜ 2 Σ u + − A ˜ 2 Π u spin–orbit coupling. Forward modeling codes of the Mars dayglow have not been accurate in the mid-UV due to systematic errors in these two emission cross sections. We furnish absolute emission cross sections for several band systems over electron energies 20–100 eV for CO2. We present a CB lifetime, which together with emission cross sections, furnish a set of fundamental physical constants for electron transport codes such as AURIC (Atmospheric Ultraviolet Radiance Integrated Code). AURIC and Trans-Mars are used in the analysis of UV spectra from the Martian dayglow and aurora.

In the parasite Trypanosoma brucei, the causative agent of human African sleeping sickness, all mRNAs are trans-spliced to generate a common 5’ exon derived from the spliced leader RNA (SL RNA). Perturbations of protein translocation across the endoplasmic reticulum (ER) induce the spliced leader RNA silencing (SLS) pathway. SLS activation is mediated by a serine-threonine kinase, PK3, which translocates from the cytosolic face of the ER to the nucleus, where it phosphorylates the TATA binding protein TRF4, leading to the shut-off of SL RNA transcription, followed by induction of programmed cell death. Here, we demonstrate that SLS is also induced by depletion of the essential ER resident chaperones BiP and calreticulin, ER oxidoreductin 1 (ERO1), and the Golgi-localized quiescin sulfhydryl oxidase (QSOX1). Most strikingly, silencing of Rhomboid-like 1(TIMRHOM1) involved in mitochondrial protein import, also induces SLS. The PK3 kinase, which integrates SLS signals, is modified by phosphorylation on multiple sites. To determine which of the phosphorylation events activate PK3, several individual mutations or their combination were generated. These mutations failed to completely eliminate the phosphorylation or translocation of the kinase to the nucleus. The structure of PK3 kinase and its ATP binding domain were therefore modeled. A conserved phenylalanine at position 771 was proposed to interact with ATP, and the PK3F771L mutation completely eliminated phosphorylation under SLS, suggesting that the activation involves most if not all the phosphorylation sites. The study suggests that the SLS occurs broadly in response to failures in protein sorting, folding, or modification across multiple compartments.

Trypanosoma brucei, the parasite that causes sleeping sickness, cycles between an insect and a mammalian host. However, the effect of RNA modifications such as pseudouridinylation on its ability to survive in these two different host environments is unclear. Here, two genome-wide approaches were applied for mapping pseudouridinylation sites (Ψs) on small nucleolar RNA (snoRNA), 7SL RNA, vault RNA, and tRNAs from T. brucei. We show using HydraPsiSeq and RiboMeth-seq that the Ψ on C/D snoRNA guiding 2'-O-methylation increased the efficiency of the guided modification on its target, rRNA. We found differential levels of Ψs on these noncoding RNAs in the two life stages (insect host and mammalian host) of the parasite. Furthermore, tRNA isoform abundance and Ψ modifications were characterized in these two life stages demonstrating stage-specific regulation. We conclude that the differential Ψ modifications identified here may contribute to modulating the function of noncoding RNAs involved in rRNA processing, rRNA modification, protein synthesis, and protein translocation during cycling of the parasite between its two hosts.

Summary The parasite Trypanosoma brucei is the causative agent of sleeping sickness and cycles between insect and mammalian hosts. The parasite appears to lack conventional transcriptional regulation of protein coding genes, and mRNAs are processed from polycistronic transcripts by the concerted action of trans-splicing and polyadenylation. Regulation of mRNA function is mediated mainly by RNA binding proteins affecting mRNA stability and translation. In this study, we describe the identification of 62 non-coding (nc) RNAs that are developmentally regulated and/or respond to stress. We characterized two novel anti-sense RNA regulators (TBsRNA-33 and 37) that originate from the rRNA loci, associate with ribosomes and polyribosomes, and interact in vivo with distinct mRNA species to regulate translation. Thus, this study suggests for the first-time anti-sense RNA regulators as an additional layer for controlling gene expression in these parasites., Graphical Abstract, Highlights • Trypanosome non-coding RNAs (ncRNAs) are developmentally regulated during cycling between two hosts • ncRNAs originate from rRNA locus and associate with the ribosome en route to cytoplasm • In vivo cross-linking enable identification of target RNA species regulated by ncRNAs • Trypanosomes possess anti-sense ncRNAs that regulate translation, Molecular Biology; Omics

NASA’s Global-scale Observation of Limb and Disk’s (GOLD) instrument observed the July 2, 2019 total solar eclipse’s effect in the thermosphere from a geostationary orbit above South America. GOLD’s observations of compositional and neutral temperature changes induced by the eclipse are different from the modeled effects. Combined Thermospheric Ionospheric Electrodynamics General Circulation Model (TIE-GCM) and GLobal airglOW (GLOW) modeling of GOLD’s observation is relatively successful in reproducing morphologically changes. However, the model underestimates the compositional changes. GOLD observation show a ΣO/N2 column density ratio enhancement of ~ 80 % near the totality, but the model predicts ~ 10 % enhancement. This indicates that there are inadequacies in current modeling capabilities for thermospheric changes during an eclipse. GOLD’s thermospheric measurements provide an important, new test of the models. We will present detailed data-model comparisons of measurements versus modeling results for the July 2nd eclipse.

The parasite Trypanosoma brucei is the causative agent of sleeping sickness, and cycles between insect and mammalian hosts. The parasite appears to lack conventional transcriptional regulation of protein coding genes, and mRNAs are processed from polycistronic transcripts by the concerted action of trans-splicing and polyadenylation. Regulation of mRNA function is mediated mainly by RNA binding proteins (RBPs) affecting mRNA stability and translation. In this study, we describe the identification of 62 non-coding (nc) RNAs that are developmentally regulated and/or respond to stress. We characterized two novel anti-sense RNA regulators (TBsRNA-33, and 37) that originate from the rRNA loci, associate with ribosomes and polyribosomes and interacts in vivo with distinct mRNA species to regulate translation. Thus, this study suggests for the first-time anti-sense RNA regulators as an additional layer for controlling gene expression in these parasites.

The parasite Trypanosoma brucei, the causative agent of sleeping sickness, cycles between an insect and a mammalian host. Here, we investigated the presence of pseudouridines (Ψs) on the spliceosomal small nuclear RNAs (snRNAs), which may enable growth at the very different temperatures characterizing the two hosts. To this end, we performed the first high-throughput mapping of spliceosomal snRNA Ψs by small RNA Ψ-seq. The analysis revealed 42 Ψs on T. brucei snRNAs, which is the highest number reported so far. We show that a trypanosome protein analogous to human protein WDR79, is essential for guiding Ψ on snRNAs but not on rRNAs. snoRNA species implicated in snRNA pseudouridylation were identified by a genome-wide approach based on ligation of RNAs following in vivo UV cross-linking. snRNA Ψs are guided by single hairpin snoRNAs, also implicated in rRNA modification. Depletion of such guiding snoRNA by RNAi compromised the guided modification on snRNA and reduced parasite growth at elevated temperatures. We further demonstrate that Ψ strengthens U4/U6 RNA–RNA and U2B"/U2A’ proteins-U2 snRNA interaction at elevated temperatures. The existence of single hairpin RNAs that modify both the spliceosome and ribosome RNAs is unique for these parasites, and may be related to their ability to cycle between their two hosts that differ in temperature.