Abstract In this study, we conducted a numerical analysis on catheter sizes using computational fluid dynamics to assess urinary flow rates during intermittent catheterization (IC). The results ...revealed that the fluid (urine) movement within a catheter is driven by intravesical pressure, with friction against the catheter walls being the main hindrance to fluid movement. Higher-viscosity fluids experienced increased friction with increasing intravesical pressure, resulting in reduced fluid velocity, whereas lower-viscosity fluids experienced reduced friction under similar pressure, leading to increased fluid velocity. Regarding urine characteristics, the results indicated that bacteriuria, with lower viscosity, exhibited higher flow rates, whereas glucosuria exhibited the lowest flow rates. Additionally, velocity gradients decreased with increasing catheter diameters, reducing friction and enhancing fluid speed, while the friction increased with decreasing diameters, reducing fluid velocity. These findings confirm that flow rates increased with larger catheter sizes. Furthermore, in terms of specific gravity, the results showed that a 12Fr catheter did not meet the ISO-suggested average flow rate (50 cc/min). The significance of this study lies in its application of fluid dynamics to nursing, examining urinary flow characteristics in catheterization. It is expected to aid nurses in selecting appropriate catheters for intermittent catheterization based on urinary test results.
HIV-1 reverse transcription (RT) occurs before or during uncoating, but the cellular compartment where RT and uncoating occurs is unknown. Using imaging and biochemical assays to track HIV-1 capsids ...in the nucleus during infection, we demonstrated that higher-order capsid complexes and/or complete cores containing the viral genome are imported into the nucleus. Inhibition of RT does not prevent capsid nuclear import; thus, RT may occur in nuclear compartments. Cytosolic and nuclear fractions of infected cells reveal that most RT intermediates are enriched in nuclear fractions, suggesting that HIV-1 RT occurs in the nucleus alongside uncoating. In agreement, we find that capsid in the nucleus induces recruitment of cleavage and polyadenylation specific factor 6 (CPSF6) to SC35 nuclear speckles, which are highly active transcription sites, suggesting that CPSF6 through capsid is recruiting viral complexes to SC35 speckles for the occurrence of RT. Thus, nuclear import precedes RT and uncoating, which fundamentally changes our understanding of HIV-1 infection.
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•Complete or slightly uncoated HIV-1 cores are imported into the nucleus•HIV-1 uncoating and reverse transcription occurs in the nucleus•CPSF6 recruits HIV-1 cores to SC35 nuclear speckles for reverse transcription
Selyutina et al. show that HIV-1 cores containing the viral genome are imported into the nucleus for reverse transcription and uncoating. HIV-1 cores in the nucleus are recruited by CPSF6 to SC35 highly active transcription domains for viral reverse transcription, integration, and/or expression.
The buildup of pressure drop with mass loading of particles aggravates the breathing resistance and energy consumption of filters. This study investigated the role of intra- and interlayer space of ...filter media on the pressure drop development with continued particle loading. Five basic morphologies, including microfibers, nanofibers, microbeads-on-strings, and a mix of those morphologies were fabricated via electrospinning. Then the variations of layered constructions were made, to include a total 14 different filter structures. For a single layer filter media, the pore size rather than the percent porosity had a major impact on the pressure drop. For dual layers, the space between the layers and the placement order of webs were important factors affecting the pressure drop and depth loading of particles. Computational modeling was used to interpret the role of the interlayer space on the pressure drop, by monitoring the air flow and particle movement within the filter constructions, where the computational prediction corresponded to the tendency of the experimental findings. The novelty of this study lies in the combined approach of the experimental and computational work to understand the particle capture phenomenon during the mass loading.
The HIV-1 genome enters cells inside a shell comprised of capsid (CA) protein. Variation in CA sequence alters HIV-1 infectivity and escape from host restriction factors. However, apart from the ...Cyclophilin A-binding loop, CA has no known interfaces with which to interact with cellular cofactors. Here we describe a novel protein-protein interface in the N-terminal domain of HIV-1 CA, determined by X-ray crystallography, which mediates both viral restriction and host cofactor dependence. The interface is highly conserved across lentiviruses and is accessible in the context of a hexameric lattice. Mutation of the interface prevents binding to and restriction by CPSF6-358, a truncated cytosolic form of the RNA processing factor, cleavage and polyadenylation specific factor 6 (CPSF6). Furthermore, mutations that prevent CPSF6 binding also relieve dependence on nuclear entry cofactors TNPO3 and RanBP2. These results suggest that the HIV-1 capsid mediates direct host cofactor interactions to facilitate viral infection.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The early events of HIV-1 infection involve the transport of the viral core into the nucleus. This event triggers the translocation of CPSF6 from paraspeckles into nuclear speckles forming ...puncta-like structures. Our investigations revealed that neither HIV-1 integration nor reverse transcription is required for the formation of puncta-like structures. Moreover, HIV-1 viruses without viral genome are competent for the induction of CPSF6 puncta-like structures. In agreement with the notion that HIV-1 induced CPSF6 puncta-like structures are biomolecular condensates, we showed that osmotic stress and 1,6-hexanediol induced the disassembly of CPSF6 condensates. Interestingly, replacing the osmotic stress by isotonic media re-assemble CPSF6 condensates in the cytoplasm of the cell. To test whether CPSF6 condensates were important for infection we utilized hypertonic stress, which prevents formation of CPSF6 condensates, during infection. Remarkably, preventing the formation of CPSF6 condensates inhibits the infection of wild type HIV-1 but not of HIV-1 viruses bearing the capsid changes N74D and A77V, which do not form CPSF6 condensates during infection
. We also investigated whether the functional partners of CPSF6 are recruited to the condensates upon infection. Our experiments revealed that CPSF5, but not CPSF7, co-localized with CPSF6 upon HIV-1 infection. We found condensates containing CPSF6/CPSF5 in human T cells and human primary macrophages upon HIV-1 infection. Additionally, we observed that the integration cofactor LEDGF/p75 changes distribution upon HIV-1 infection and surrounds the CPSF6/CPSF5 condensates. Overall, our work demonstrated that CPSF6 and CPSF5 are forming biomolecular condensates that are important for infection of wild type HIV-1 viruses.
Lentiviruses such as HIV-1 traverse nuclear pore complexes (NPC) and infect terminally differentiated non-dividing cells, but how they do this is unclear. The cytoplasmic NPC protein Nup358/RanBP2 ...was identified as an HIV-1 co-factor in previous studies. Here we report that HIV-1 capsid (CA) binds directly to the cyclophilin domain of Nup358/RanBP2. Fusion of the Nup358/RanBP2 cyclophilin (Cyp) domain to the tripartite motif of TRIM5 created a novel inhibitor of HIV-1 replication, consistent with an interaction in vivo. In contrast to CypA binding to HIV-1 CA, Nup358 binding is insensitive to inhibition with cyclosporine, allowing contributions from CypA and Nup358 to be distinguished. Inhibition of CypA reduced dependence on Nup358 and the nuclear basket protein Nup153, suggesting that CypA regulates the choice of the nuclear import machinery that is engaged by the virus. HIV-1 cyclophilin-binding mutants CA G89V and P90A favored integration in genomic regions with a higher density of transcription units and associated features than wild type virus. Integration preference of wild type virus in the presence of cyclosporine was similarly altered to regions of higher transcription density. In contrast, HIV-1 CA alterations in another patch on the capsid surface that render the virus less sensitive to Nup358 or TRN-SR2 depletion (CA N74D, N57A) resulted in integration in genomic regions sparse in transcription units. Both groups of CA mutants are impaired in replication in HeLa cells and human monocyte derived macrophages. Our findings link HIV-1 engagement of cyclophilins with both integration targeting and replication efficiency and provide insight into the conservation of viral cyclophilin recruitment.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
In the outbreak of COVID-19, the extended wear of single-use, disposable respirators was inevitable due to limited supplies. As a respirator is front-line protection against particulate matter, ...including bioaerosol and droplets, a comprehensive understanding for the reuse strategy is needed. In this study, eight different disinfection methods commonly applied for the reuse of respirators were compared for their influence on the filtration and bactericidal/bacteria removal performance, with in-depth discussion on the cause of effects. Treatments including oven-dry, ultraviolet irradiation (UV), microwaving, laundering with and without detergent, and immersion in hypochlorite, isopropanol, and ethanol were performed to respirators. Immersion in ethanol or isopropanol was effective for inactivation and removal of bacteria, yet such a treatment significantly deteriorated the filtration efficiency in about 20-28%, dissipating the surface charges. Laundering, while effective in removing the attached bacteria, triggered physical damage, leading to a possible reduction of filtration performance. A short-term oven-dry, UV irradiation, and microwaving mostly preserved the filtration performance, yet the drawback lied in the incomplete bactericidal efficiency. This study would contribute to the public health and safety by providing scientific background on the effect of disinfection treatment methods for respirators.
The movement of viruses and other large macromolecular cargo through nuclear pore complexes (NPCs) is poorly understood. The human immunodeficiency virus type 1 (HIV-1) provides an attractive model ...to interrogate this process. HIV-1 capsid (CA), the chief structural component of the viral core, is a critical determinant in nuclear transport of the virus. HIV-1 interactions with NPCs are dependent on CA, which makes direct contact with nucleoporins (Nups). Here we identify Nup35, Nup153, and POM121 to coordinately support HIV-1 nuclear entry. For Nup35 and POM121, this dependence was dependent cyclophilin A (CypA) interaction with CA. Mutation of CA or removal of soluble host factors changed the interaction with the NPC. Nup35 and POM121 make direct interactions with HIV-1 CA via regions containing phenylalanine glycine motifs (FG-motifs). Collectively, these findings provide additional evidence that the HIV-1 CA core functions as a macromolecular nuclear transport receptor (NTR) that exploits soluble host factors to modulate NPC requirements during nuclear invasion.
Transportin 3 (Tnpo3, Transportin-SR2) is implicated in nuclear import of splicing factors and HIV-1 replication. Herein, we show that the majority of cellular Tnpo3 binding partners contain ...arginine-serine (RS) repeat domains and present crystal structures of human Tnpo3 in its free as well as GTPase Ran- and alternative splicing factor/splicing factor 2 (ASF/SF2)-bound forms. The flexible β-karyopherin fold of Tnpo3 embraces the RNA recognition motif and RS domains of the cargo. A constellation of charged residues on and around the arginine-rich helix of Tnpo3 HEAT repeat 15 engage the phosphorylated RS domain and are critical for the recognition and nuclear import of ASF/SF2. Mutations in the same region of Tnpo3 impair its interaction with the cleavage and polyadenylation specificity factor 6 (CPSF6) and its ability to support HIV-1 replication. Steric incompatibility of the RS domain and RanGTP engagement by Tnpo3 provides the mechanism for cargo release in the nucleus. Our results elucidate the structural bases for nuclear import of splicing factors and the Tnpo3–CPSF6 nexus in HIV-1 biology.
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