The evolutionary conserved TLDc domain defines a new class of (H+)V-ATPase interacting proteins

We recently found that nuclear receptor coactivator 7 (Ncoa7) and Oxr1 interact with the proton-pumping V-ATPase. Ncoa7 and Oxr1 belong to a group of proteins playing a role in the oxidative stress response, that contain the conserved “TLDc” domain. Here we asked if the three other proteins in this family, i.e., Tbc1d24, Tldc1 and Tldc2 also interact with the V-ATPase and if the TLDc domains are involved in all these interactions. By co-immunoprecipitation, endogenous kidney Tbc1d24 (and Ncoa7 and Oxr1) and overexpressed Tldc1 and Tldc2, all interacted with the V-ATPase. In addition, purified TLDc domains of Ncoa7, Oxr1 and Tldc2 (but not Tbc1d24 or Tldc1) interacted with V-ATPase in GST pull-downs. At the amino acid level, point mutations G815A, G845A and G896A in conserved regions of the Ncoa7 TLDc domain abolished interaction with the V-ATPase, and S817A, L926A and E938A mutations resulted in decreased interaction. Furthermore, poly-E motifs upstream of the TLDc domain in Ncoa7 and Tldc2 show a (nonsignificant) trend towards enhancing the interaction with V-ATPase. Our principal finding is that all five members of the TLDc family of proteins interact with the V-ATPase. We conclude that the TLDc motif defines a new class of V-ATPase interacting regulatory proteins.

The Evolutionary Conserved TLDc Domain Defines a New Class of (H+)V-ATPase Interacting Proteins

We recently found that nuclear receptor coactivator 7 (Ncoa7) and Oxr1 interact with the proton-pumping V-ATPase. Ncoa7 and Oxr1 belong to a group of proteins playing a role in the oxidative stress response, that contain the conserved “TLDc” domain. Here we asked if the three other proteins in this family, i.e., Tbc1d24, Tldc1 and Tldc2 also interact with the V-ATPase and if the TLDc domains are involved in all these interactions. By co-immunoprecipitation, endogenous kidney Tbc1d24 (and Ncoa7 and Oxr1) and overexpressed Tldc1 and Tldc2, all interacted with the V-ATPase. In addition, the purified TLDc domains of Ncoa7, Oxr1 and Tldc2 (but not Tbc1d24 or Tldc1) interacted with V-ATPase in GST pull-downs. At the amino acid level, the point mutations G815A, G845A and G896A in conserved regions of the Ncoa7 TLDc domain abolished interaction with the V-ATPase, and S817A, L926A and E938A mutations resulted in decreased interaction. Furthermore, poly-E motifs present upstream of the TLDc domain in Ncoa7 and Tldc2 enhancedshow a (nonsignificant) trend towards enhancing the interaction with V-ATPase. Our principal finding is that all five members of the TLDc family of proteins interact with the V-ATPase. We conclude that the TLDc motif defines a new class of V-ATPase interacting regulatory proteins.

Serum IL-18 Is a Potential Biomarker for Predicting Severe Dengue Disease Progression

Background. Dengue virus (DENV) infection is the most common arboviral disease that affects tropical and subtropical regions. Based on the clinical hallmarks, the different severities of patients range from mild dengue fever (MDF) to severe dengue diseases (SDDs) and include dengue hemorrhagic fever or dengue shock syndrome. These are commonly associated with cytokine release syndrome (CRS). The types and levels of cytokines/chemokines, which are suppressed or enhanced, are varied, indicating CRS’s pathogenic and host defensive effects. Principal Finding. In this study, we created an integrated and precise multiplex panel of cytokine/chemokine assays based on our literature analysis to monitor dengue CRS. A 24-plex panel of cytokines/chemokines was evaluated to measure the plasma levels of targeting factors in dengue patients with an MDF and SDD diagnosis without or with comorbidities. As identified in sixteen kinds of cytokines/chemokines, ten were significantly ( P < 0.05 ) (10/16) increased, one was significantly ( P < 0.01 ) (1/16) decreased, and five were potentially (5/16) altered in all dengue patients ( n = 30 ) in the acute phase of disease onset. Compared to MDF, the levels of IL-8 (CXCL-8) and IL-18 in SDD were markedly ( P < 0.05 ) increased, accompanied by positively increased IL-6 and TNF-α and decreased IFN-γ and RANTES. With comorbidities, SDD significantly ( P < 0.01 ) portrayed elevated IL-18 accompanied by increased IL-6 and decreased IFN-α2 and IL-12. In addition, decreased platelets were significantly ( P < 0.05 ) associated with increased IL-18. Significance. These results demonstrate an efficient panel of dengue cytokine/chemokine assays used to explore the possible level of CRS during the acute phase of disease onset; also, we are the first to report the increase of IL-18 in severe dengue with comorbidity compared to severe dengue without comorbidity and mild dengue.

Access to Government Information and Corruption: An Empirical Investigation

This study taps into a newly constructed index of actual access to government information with a view to finding out whether such access reduces corruption. It draws on a few cases at the micro level and on cross-country data at the macro level to investigate this research question. The study is designed to detect for methodological problems commonly encountered in regression models used to empirically investigate the causes of corruption. The principal finding of the study is that access to government information reduces corruption at both the micro and macro levels. Another key finding is that a supporting institutional and political environment is necessary for the effectiveness of access to government information as an anti-corruption policy tool. A third finding is that access to government information through the traditional is an effective policy tool for curbing corruption, particularly in contexts characterized by limited computer and Internet penetration in society. This study underscores the importance of using traditional media as civic engagement tools in anti-corruption policy initiatives. The policy implication here is that access to government information through the traditional media may have to be an important component of anti-corruption policy initiatives.

Risk of Trypanosoma cruzi infection among travellers visiting friends and relatives to continental Latin America

Background Chagas disease (CD) is regarded as a possible risk for travellers to endemic areas of continental Latin America (LA). The aim of the study is to determine the risk of Trypanosoma cruzi (TC) infection among travellers to CD endemic areas and to identify risk factors for acquiring TC infection. Methods/Principal finding We designed a multicenter cross-sectional study among travellers in Spain (Badalona, Barcelona and Madrid). All available adults with laboratory confirmed proof of absence of TC infection from January 2012 to December 2015 were contacted. Participants referring a trip to LA after the negative TC screening were offered to participate. We performed a standardized questionnaire of travel related factors and measurement of TC antibodies in serum. A total of 971 participants with baseline negative TC serology were selected from the microbiology records. After excluding participants not meeting inclusion criteria, eighty participants were selected. Sixty three (78.8%) were female, and the median age was 38 (IQR 34–47) years. The reason to travel was visiting friends and relatives in 98.8% of the participants. The median duration of travel was 40 (IQR 30–60) days, with 4911 participants-day of exposure. Seventy seven cases (96.25%) participants had two negative TC serology tests after the travel, two cases (2.5%) had discordant serology results (considered false positive results) and one case was infected before travelling to LA. According to our data, the upper limit of the 95% confidence interval of the incidence rate of TC acquisition in travellers is 0.8 per 1000 participant-days. Conclusions/Significance Among 79 non-CD travellers to TC endemic areas, we found no cases of newly acquired TC infection. The incidence rate of TC acquisition in travellers to endemic countries is less than or equal to 0.8 per 1000 traveller-days.

An atomistic model of the coronavirus replication-transcription complex as a hexamer assembled around nsp15

Using available cryo-EM and x-ray crystal structures of the nonstructural proteins that are responsible for SARS-CoV-2 viral RNA replication and transcription, we have constructed an atomistic model of how the proteins assemble into a functioning superstructure. Our principal finding is that the complex is hexameric, centered around nsp15. The nsp15 hexamer is capped on two faces by trimers of nsp14/nsp16/(nsp10) 2 , where nsp14 is seen to undergo a large conformational change between its two domains. This conformational change facilitates binding of six nsp12/nsp7/(nsp8) 2 polymerase subunits to the complex. To this, six subunits of nsp13 are arranged around the superstructure, but not evenly distributed. Two of the six polymerase subunits are each proposed to carry dimers of nsp13, while two others are proposed to carry monomers. The polymerase subunits that coordinate nsp13 dimers also bind the nucleocapsid, which positions the 5’-UTR TRS-L RNA over the polymerase active site, a state distinguishing transcription from replication. Analyzing the path of the viral RNA indicates the dsRNA that exits the polymerase passes over the nsp14 exonuclease and nsp15 endonuclease sites before being unwound by a convergence of zinc fingers from nsp10 and nsp14. The template strand is then directed away from the complex, while the nascent strand is directed to the sites responsible for mRNA capping (the nsp12 NiRAN and the nsp14 and nsp16 methyltransferases). The model presents a cohesive picture of the multiple functions of the coronavirus replication-transcription complex and addresses fundamental questions related to proofreading, template switching, mRNA capping and the role of the endonuclease. It provides a platform to guide biochemical and structural research to address the stoichiometric and spatial configuration of the replication-transcription complex.

A Mathematical Model of the Rat Kidney. III. Ammonia Transport

Ammonia generated within the kidney is partitioned into a urinary fraction (the key buffer for net acid excretion), and an aliquot delivered to the systemic circulation. The physiology of this partitioning has yet to be examined in a kidney model, and that is undertaken in this work. This involves explicit representation of the cortical labyrinth, so that cortical interstitial solute concentrations are computed, rather than assigned. A detailed representation of cortical vasculature has been avoided by making the assumption that solute concentrations within interstitium and peritubular capillaries are likely to be identical, and that there is little to no modification of venous composition as blood flows to the renal vein. The model medullary ray has also been revised to include a segment of proximal straight tubule, which supplies ammonia to this region. The principal finding of this work is that cortical labyrinth interstitial ammonia concentration is likely to be several-fold higher than systemic arterial ammonia. This elevation of interstitial ammonia enhances ammonia secretion in both PCT and DCT, with uptake by Na,K-ATPases of both segments. Model prediction of urinary ammonia excretion is concordant with measured values, but at the expense of greater ammoniagenesis, with high rates of renal venous ammonia flux. This derives from a limited capability of the model medulla to replicate the high interstitial ammonia concentrations that are required to drive collecting duct ammonia secretion. Thus, renal medullary ammonia trapping appears key to diverting ammonia from renal vein to urine, but capturing the underlying physiology remains a challenge.

Realism and Naturalism in Samrat Upadhyay’s The Guru of Love

This research project seeks to explore realism and naturalism in Samrat Upadhyay's novel, The Guru of Love, in the context of Nepalese society. It analyses how the novel delineates a realistic account of Nepalese middle-class people's hardships, struggles, and problems they face internally and externally through the character of Ramchandra, the protagonist of the fiction, an ordinary mathematics teacher who faces some hindrances in search of a better life and developing a career in the capital city. The principal objective of the study is to find the realistic, naturalistic, and materialistic elements in the novel, to examine the psychic predispositions that characters come through realistically, and to analyze the role of love and sex in terms of spirituality and emancipation as they all fall into the parameters of naturalism and realism. This study is developed through a theoretical framework based on realism and naturalism for the close textual analysis of the novel. The principal finding of the research paper is that the major characters of the novel, The Guru of Love, suffer from the conflict between their desires and their socio-economic realities. The article also surveys how material prosperity fails to resolve all the problems that human being goes through. It is expected that the paper stands as a reference for the research scholars interested to explore in the area.

Pentacyclic Triterpene Profile and Its Biosynthetic Pathway in Cecropia telenitida as a Prospective Dietary Supplement

Promising research over the past decades has shown that some types of pentacyclic triterpenes (PTs) are associated with the prevention of type 2 diabetes (T2D), especially those found in foods. The most abundant edible sources of PTs are those belonging to the ursane and oleanane scaffold. The principal finding is that Cecropia telenitida contains abundant oleanane and ursane PT types with similar oxygenation patterns to those found in food matrices. We studied the compositional profile of a rich PT fraction (DE16-R) and carried out a viability test over different cell lines. The biosynthetic pathway connected to the isolated PTs in C. telenitida offers a specific medicinal benefit related to the modulation of T2D. This current study suggests that this plant can assemble isobaric, positional isomers or epimeric PT. Ursane or oleanane scaffolds with the same oxygenation pattern are always shared by the PTs in C. telenitida, as demonstrated by its biosynthetic pathway. Local communities have long used this plant in traditional medicine, and humans have consumed ursane and oleanane PTs in fruits since ancient times, two key points we believe useful in considering the medicinal benefits of C. telenitida and explaining how a group of molecules sharing a closely related scaffold can express effectiveness.

Characterisation of V-type asteroids orbiting in the middle and outer main belt

We present new spectral observations using ground-based telescopes of 23 putative V-type asteroids, selected according to colour surveys in the visible from the Moving Objects Catalogue of the Sloan Digital Sky Survey and near-infrared from the Moving Objects VISTA catalogue. Ten asteroids are orbiting in the middle main belt, while five in the outer part of the main asteroid belt. For the observed asteroids we assign a taxonomical classification and confirm the basaltic nature for sixteen of them. The high-quality spectra in the UV range, obtained with the X-Shooter spectrograph at ESO, allowed the identification of the Fe2 + forbidden transition of pyroxene for ten asteroids. This band is centred at 506.5 nm, and it is diagnostic of the Ca-content in the pyroxene form. We determined a low Fe-content composition for asteroids (2452) Lyot, (5758) Brunini, (7675) Gorizia, (9197) Endo, (22308) 1990 UO4, (36118) 1999 RE135, (66905) 1999 VC160, and (189597) 2000 WG119, and a composition more rich in Fe for asteroids (75661) 2000 AB79 and (93620) 2000 UQ70. We also present a dynamical investigation of V-type asteroids in the middle and outer main belt. The principal finding of these simulations is that the middle and outer V-types are more likely to be associated with some families, which were considered as possibly originated from the break-up of a partially or totally differentiated parent body by diverse studies. This reinforces the hypothesis that the identified V-type in the region were not originated from (4) Vesta and that the number of differentiated objects in the middle and outer main belt must have been much larger than previously assumed.