Dynamics and Patterning of 5-Hydroxytryptamine 2 Subtype Receptors in JC Polyomavirus Entry.

The organization and dynamics of plasma membrane receptors are a critical link in virus-receptor interactions, which finetune signaling efficiency and determine cellular responses during infection. Characterizing the mechanisms responsible for the active rearrangement and clustering of receptors may aid in developing novel strategies for the therapeutic treatment of viruses. Virus-receptor interactions are poorly understood at the nanoscale, yet they present an attractive target for the design of drugs and for the illumination of viral infection and pathogenesis. This study utilizes super-resolution microscopy and related techniques, which surpass traditional microscopy resolution limitations, to provide both a spatial and temporal assessment of the interactions of human JC polyomavirus (JCPyV) with 5-hydroxytrypamine 2 receptors (5-HT2Rs) subtypes during viral entry. JCPyV causes asymptomatic kidney infection in the majority of the population and can cause fatal brain disease, and progressive multifocal leukoencephalopathy (PML), in immunocompromised individuals. Using Fluorescence Photoactivation Localization Microscopy (FPALM), the colocalization of JCPyV with 5-HT2 receptor subtypes (5-HT2A, 5-HT2B, and 5-HT2C) during viral attachment and viral entry was analyzed. JCPyV was found to significantly enhance the clustering of 5-HT2 receptors during entry. Cluster analysis of infected cells reveals changes in 5-HT2 receptor cluster attributes, and radial distribution function (RDF) analyses suggest a significant increase in the aggregation of JCPyV particles colocalized with 5-HT2 receptor clusters in JCPyV-infected samples. These findings provide novel insights into receptor patterning during viral entry and highlight improved technologies for the future development of therapies for JCPyV infection as well as therapies for diseases involving 5-HT2 receptors.

The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth.

A tobacco chloroplast hypothetical open reading frame 4 (YCF4) has been reported as a non-essential assembly factor for photosynthesis based on an incomplete knockout of YCF4, just 93 of 184 amino acids from the N-terminus were knocked out. On the other hand, we removed the complete sequence of YCF4 from tobacco chloroplasts and observed that ΔYCF4 plants were unable to survive photoautotrophically as their growth was hampered in the absence of an external carbon supply, clearly showing that the YCF4 is essential for photosynthesis. Initially, the aadA gene was introduced into the tobacco plastome replacing the complete YCF4 gene through homologous recombination events. The replacement of YCF4 with aadA was confirmed by PCR and Southern blot analysis in ΔYCF4 plants. Homoplasmic ΔYCF4 plants had a light green phenotype, and the leaves became pale yellow as the plants grew older. The structure of chloroplasts of ΔYCF4 mutants of light green phenotype was studied using a transmission electron microscope (TEM), and the micrographs demonstrated structural anomalies in the chloroplasts; including shape, size, and grana stacking compared to the wild-type plants. Further, transcriptome analysis revealed that the expression of PSI, PSII, and ribosomal genes remained unchanged in ∆YCF4 plants. On the other hand, transcriptome levels of rbcL (Ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit), LHC (Light-Harvesting Complex), and ATP Synthase (atpB and atpL) decreased, indicating that the YCF4 has the function(s) in addition to assembling the photosynthetic complex. This was confirmed by in-silico protein-protein interactions of full-length YCF4 as well as 93 and 91 of 184 amino acids from N- and C-termini of the full-length protein, which revealed that the C-terminus (91 aa) of YCF4 is important in interacting with other chloroplast proteins. These findings provide genetic support for the plastid YCF4 gene's critical role in regulating the plastid gene expression and assembling the photosynthetic complex.

Mechanism and Effects of Cellular Creep in a Microfluidic Filter.

Biomicroparticles such as proteins, bacterium, and cells are known to be viscoelastic, which significantly affects their performance in microfluidic applications. However, the exact effects and the quantitative study of cellular viscoelastic creep within different applications remain unclear. In this study, the cellular-deforming evolution within a filter unit was studied using a multiphysics numerical model. A general cellular creep deformation process of viscoelastic particle trapping in pores was revealed. Two featured variables, namely, the maximum surface displacement and the volumetric strain, were identified and determined to quantitatively describe the evolution. The effects of flow conditions and physical characteristics of the microparticles were studied. Furthermore, a Giardia concentration experiment was conducted using an integrated hydraulic filtration system with a porous membrane. The experimental results agreed well with the numerical analysis, indicating that, compared to pure elastic particles, it is more difficult to release cellular material matters including cells, chemical synthetic particles, and microbes from trapping due to their time-accumulated creep deformation.

Microstructure and Corrosion Behavior of Atmospheric Plasma Sprayed NiCoCrAlFe High Entropy Alloy Coating.

High entropy alloys (HEAs) are multi-elemental alloy systems that exhibit a combination of exceptional mechanical and physical properties, and nowadays are validating their potential in the form of thermal sprayed coatings. In the present study, a novel synthesis method is presented to form high entropy alloy coatings. For this purpose, thermal sprayed coatings were deposited on Stainless Steel 316L substrates using atmospheric plasma spraying technique with subsequent annealing, at 1000 °C for 4 h, to assist alloy formation by thermal diffusion. The coatings in as-coated samples as well as in annealed forms were extensively studied by SEM for microstructure and cross-sectional analysis. Phase identification was performed by X-ray diffraction studies. The annealed coatings revealed a mixed BCC and FCC based HEA structure. Potentiodynamic corrosion behavior of SS316L sprayed as well as annealed coatings were also carried out in 3.5% NaCl solution and it was found that the HEA-based annealed coatings displayed the best corrosion resistance 0.83 (mpy), as compared to coated/non-annealed and SS 316 L that showed corrosion resistance of 7.60 (mpy) and 3.04 (mpy), respectively.

Competency of groundwater recharge of irrigated cotton field subjacent to sowing methods, plastic mulch, water productivity, and yield under climate change.

Irrigated agriculture is a foremost consumer of water resources to fulfill the demand for food and fiber with an increasing population under climate changes; cotton is no exception. Depleting groundwater recharge and water productivity is critical for the sustainable cotton crop yield peculiarly in the semiarid region. This study investigated the water productivity and cotton yield under six different treatments: three sowing methods, i.e., flat, ridge, and bed planting with and without plastic mulch. Cotton bed planting without mulch showed maximum water productivity (0.24 kg.m-3) and the highest cotton yield (1946 kg.ha-1). Plastic mulching may reduce water productivity and cotton yield. HYDRUS-1D unsaturated flow model was used to access the groundwater recharge for 150 days under six treatments after model performance evaluation. Maximum cumulative recharge was observed 71 cm for the flat sowing method without plastic mulch. CanESM2 was used to predict climate scenarios for RCP 2.6, 4.5, and 8.5 for the 2050s and 2080s by statistical downscale modeling (SDSM) using historical data from 1975 to 2005 to access future groundwater recharge flux. Average cumulative recharge flux declined 36.53% in 2050 and 22.91% in 2080 compared to 2017 without plastic mulch. Multivariate regression analysis revealed that a maximum 23.78% reduction in groundwater recharge could influence future climate change. Further study may require to understand the remaining influencing factor of depleting groundwater recharge. Findings highlight the significance of climate change and the cotton sowing method while accessing future groundwater resources in irrigated agriculture.

Enhanced visible light photocatalytic activity of TiO2 co-doped with Fe, Co, and S for degradation of Cango red.

The photocatalytic activity of TiO2 was enhanced under visible light when it was co-doped with Fe, Co, and S through the sol-gel method. The resultant nano photocatalysts were characterized by XRD, EDX coupled with SEM, FT-IR, UV-Vis, and UV-DRS. The concentration ratio of precursor salts of Fe and S was kept at 1% each and of Co varied between 0.5% -1.5%. The photodegradation of Congo red was carried out and various factors like the concentration of catalyst, initial concentration of dye, pH, and irradiation time were also investigated to optimize the degradation process. Under optimized degradation conditions, 99.3% of Congo red (30 ppm) was degraded at slightly acidic pH, with 0.14 g of photocatalyst within 70 min of irradiation time.

Sending mixed signals: polyomavirus entry and trafficking.

Polyomaviruses are mostly non-pathogenic, yet some can cause human disease especially under conditions of immunosuppression, including JC, BK, and Merkel cell polyomaviruses. Direct interactions between viruses and the host early during infection dictate the outcome of disease, many of which remain enigmatic. However, significant work in recent years has contributed to our understanding of how this virus family establishes an infection, largely due to advances made for animal polyomaviruses murine and SV40. Here we summarize the major findings that have contributed to our understanding of polyomavirus entry, trafficking, disassembly, signaling, and immune evasion during the infectious process and highlight major unknowns in these processes that are open areas of study.

Electrochemical Properties of Tin Sulfide Nano-Sheets as Cathode Material for Lithium-Sulfur Batteries.

Unprecedented self-assembled hierarchical nano-sheets of SnS were synthesized by the hydrothermal method. In a typical reaction, SnCl2.2H2O and Na2S.9H2O were used as reactants. Structural and morphological properties were studied by X-ray diffraction (XRD), and scanning electron microscopy (SEM) while the electrochemical properties were measured by cyclic voltammetry, charge-discharge cycles, and electrochemical impedance spectroscopy (EIS). SEM results showed the 1-D SnS nano-sheets with an average thickness of around 20 nm. Cyclic voltammogram and charge-discharge spectra showed good cycling stability. All these results showed that SnS nano-sheets are promising candidate material to be used as electrode for Li-S batteries.

Influenza Hemagglutinin Modulates Phosphatidylinositol 4,5-Bisphosphate Membrane Clustering.

The lipid phosphatidylinositol 4,5-bisphosphate (PIP2) forms nanoscopic clusters in cell plasma membranes; however, the processes determining PIP2 mobility and thus its spatial patterns are not fully understood. Using super-resolution imaging of living cells, we find that PIP2 is tightly colocalized with and modulated by overexpression of the influenza viral protein hemagglutinin (HA). Within and near clusters, HA and PIP2 follow a similar spatial dependence, which can be described by an HA-dependent potential gradient; PIP2 molecules move as if they are attracted to the center of clusters by a radial force of 0.079 ± 0.002 pN in HAb2 cells. The measured clustering and dynamics of PIP2 are inconsistent with the unmodified forms of the raft, tether, and fence models. Rather, we found that the spatial PIP2 distributions and how they change in time are explained via a novel, to our knowledge, dynamic mechanism: a radial gradient of PIP2 binding sites that are themselves mobile. This model may be useful for understanding other biological membrane domains whose distributions display gradients in density while maintaining their mobility.

Dupatta (Long Scarf) related injuries in female pillion riders in Karachi Pakistan.

OBJECTIVE: To assess the frequency, type of injuries and reason behind avoidable injuries due to dupatta entrapment in motorcycle wheels among female pillion riders. METHODS: This observational / descriptive study was conducted at the Jinnah Postgraduate Medical Centre, Karachi, from July 2013 to December 2014, and comprised female patients with dupatta-related motorbike injuries. The patients had injuries around head and neck, face, chest, abdomen, etc. SPSS 11 was used for data analysis. RESULTS: Of the 450 dupatta-related injuries, 49(10.89%) patients were admitted, whereas the remainder with minor injuries on neck and other parts of body were primarily managed in the accident and emergency department, detained for observation and discharged after first aid. The mean age was 31.5±19.09 years (range: 18 to 45 years). Besides, 37(75.51%) patients were admitted to the orthopaedic department, 5(10.2%) to the ear, nose and throat department, 4(8.16%) to the neurosurgery department and 2(4.08%) to the general surgery department. The number of injuries related to head and neck was 11(23%), face 8(17%), chest, abdomen and pelvic contents 1(2%) each, extremity and pelvic girdle 14(29%) and external injuries 13(28%). CONCLUSIONS: Dupatta entrapment was found to be a common cause of motorcycle accidents.

Inhibition of GLI1 Expression by Targeting the CRD-BP-GLI1 mRNA Interaction Using a Specific Oligonucleotide.

The stabilization of glioma-associated oncogene 1 (GLI1) mRNA by coding region determinant binding protein (CRD-BP) through the Wnt/ß-catenin signaling pathway is implicated in the proliferation of colorectal cancer and basal cell carcinoma. Here, we set out to characterize the physical interaction between CRD-BP and GLI1 mRNA so as to find inhibitors for such interaction. Studies using CRD-BP variants with a point mutation in the GXXG motif at each KH domain showed that KH1 and KH2 domain are critical for the binding of GLI1 RNA. The smallest region of GLI1 RNA binding to CRD-BP was mapped to nucleotides (nts) 320-380. A 37-nt S1 RNA sense oligonucleotide, containing two distinct stem-loops present in nts 320-380 of GLI1 RNA, was found to be effective in blocking CRD-BP-GLI1 RNA interaction. Studies using various competitor RNAs with modifications to S1 RNA oligonucleotide further displayed that both the sequences and the structure of the two stem-loops are important for CRD-BP-GLI1 RNA binding. The role of the two-stem-loop motif in influencing CRD-BP-RNA interaction was further investigated in cells. The 2'-O-methyl derivative of the S1 RNA oligonucleotide significantly decreased GLI1, c-myc, and CD44 mRNA levels, in a panel of colon and breast cancer cells. The results from this study demonstrate the potential importance of the two-stem-loop motif as a target region for the inhibition of the CRD-BP-GLI1 RNA interaction and Hedgehog signaling pathway. Such results pave the way for the development of novel inhibitors that act by destabilizing the CRD-BP-GLI1 mRNA interaction.

Molecular insights into the coding region determinant-binding protein-RNA interaction through site-directed mutagenesis in the heterogeneous nuclear ribonucleoprotein-K-homology domains.

The ability of its four heterogeneous nuclear RNP-K-homology (KH) domains to physically associate with oncogenic mRNAs is a major criterion for the function of the coding region determinant-binding protein (CRD-BP). However, the particular RNA-binding role of each of the KH domains remains largely unresolved. Here, we mutated the first glycine to an aspartate in the universally conserved GXXG motif of the KH domain as an approach to investigate their role. Our results show that mutation of a single GXXG motif generally had no effect on binding, but the mutation in any two KH domains, with the exception of the combination of KH3 and KH4 domains, completely abrogated RNA binding in vitro and significantly retarded granule formation in zebrafish embryos, suggesting that any combination of at least two KH domains cooperate in tandem to bind RNA efficiently. Interestingly, we found that any single point mutation in one of the four KH domains significantly impacted CRD-BP binding to mRNAs in HeLa cells, suggesting that the dynamics of the CRD-BP-mRNA interaction vary over time in vivo. Furthermore, our results suggest that different mRNAs bind preferentially to distinct CRD-BP KH domains. The novel insights revealed in this study have important implications on the understanding of the oncogenic mechanism of CRD-BP as well as in the future design of inhibitors against CRD-BP function.