Visual outcomes of the surgical rehabilitative process following open globe injury repair

BackgroundThe path of rehabilitation of an eye after open globe injury (OGI) may require multiple additional secondary surgeries after the initial repair. Although much has been studied regarding the outcomes of secondary surgeries after open globe repair, it can be challenging to understand the possible implications of the surgical rehabilitative process. This retrospective study considers the benefits of the required additional secondary surgeries for a consecutive series of OGI patients.MethodsOGI patients who had at least one additional surgery after the initial open globe repair (OGR) were studied retrospectively. Additional inclusion criteria included: follow up of at least 12 months since the initial injury and at least 3 months since their most recent surgery, and no additional planned interventions. Preoperative visual acuity was compared to final visual acuity. Additionally, the odds of achieving ambulatory vision (≥20/800) and reading vision (≥20/40) were calculated after each indicated consecutive surgery.ResultsA cohort of 74 eyes from 73 patients met our inclusion criteria. These patients underwent a mean of two additional surgeries. The mean logMAR VA improved from 2.3 (HM) at presentation to 1.4 (20/150), or a 9-line Snellen equivalent improvement. Upon reaching their final visit status, 50% of patients had achieved ambulatory vision and 30% of patients had achieved reading vision. The odds of achieving ambulatory vision after completion of all the rehabilitative surgical process compared to the vision prior to the secondary rehabilitative surgery were higher (OR: 19.1, 95% CI: 7.9 – 30.4, p = 0.0008) as were the odds of achieving reading vision (OR: 4.6, 95% CI: 0.2 – 9.0, p = 0.04). With subsequent second, third, and fourth additional surgeries, the odds of achieving either ambulatory or reading vision at the final visit compared to their preoperative visual acuities were not significant (p > 0.05) but the visual acuity continued to trend toward visual improvement.ConclusionApproximately 50% of individuals who required additional surgery at UMN achieved ambulatory vision and 30% achieved reading vision. The odds of visual improvement through the surgical rehabilitative process were very high, with the greatest gains generally achieved after the first surgery

A global experiment on motivating social distancing during the COVID-19 pandemic

Finding communication strategies that effectively motivate social distancing continues to be a global public health priority during the COVID-19 pandemic. This cross-country, preregistered experiment (n = 25,718 from 89 countries) tested hypotheses concerning generalizable positive and negative outcomes of social distancing messages that promoted personal agency and reflective choices (i.e., an autonomy-supportive message) or were restrictive and shaming (i.e., a controlling message) compared with no message at all. Results partially supported experimental hypotheses in that the controlling message increased controlled motivation (a poorly internalized form of motivation relying on shame, guilt, and fear of social consequences) relative to no message. On the other hand, the autonomy-supportive message lowered feelings of defiance compared with the controlling message, but the controlling message did not differ from receiving no message at all. Unexpectedly, messages did not influence autonomous motivation (a highly internalized form of motivation relying on one’s core values) or behavioral intentions. Results supported hypothesized associations between people’s existing autonomous and controlled motivations and self-reported behavioral intentions to engage in social distancing. Controlled motivation was associated with more defiance and less long-term behavioral intention to engage in social distancing, whereas autonomous motivation was associated with less defiance and more short- and long-term intentions to social distance. Overall, this work highlights the potential harm of using shaming and pressuring language in public health communication, with implications for the current and future global health challenges

Para-infectious brain injury in COVID-19 persists at follow-up despite attenuated cytokine and autoantibody responses

To understand neurological complications of COVID-19 better both acutely and for recovery, we measured markers of brain injury, inflammatory mediators, and autoantibodies in 203 hospitalised participants; 111 with acute sera (1–11 days post-admission) and 92 convalescent sera (56 with COVID-19-associated neurological diagnoses). Here we show that compared to 60 uninfected controls, tTau, GFAP, NfL, and UCH-L1 are increased with COVID-19 infection at acute timepoints and NfL and GFAP are significantly higher in participants with neurological complications. Inflammatory mediators (IL-6, IL-12p40, HGF, M-CSF, CCL2, and IL-1RA) are associated with both altered consciousness and markers of brain injury. Autoantibodies are more common in COVID-19 than controls and some (including against MYL7, UCH-L1, and GRIN3B) are more frequent with altered consciousness. Additionally, convalescent participants with neurological complications show elevated GFAP and NfL, unrelated to attenuated systemic inflammatory mediators and to autoantibody responses. Overall, neurological complications of COVID-19 are associated with evidence of neuroglial injury in both acute and late disease and these correlate with dysregulated innate and adaptive immune responses acutely

Pten-Null Tumors Cohabiting the Same Lung Display Differential AKT Activation and Sensitivity to Dietary Restriction

PTEN loss is considered a biomarker for activated phosphoinositide 3-kinase (PI3K)/AKT, a pathway frequently mutated in cancer, and was recently shown to confer resistance to dietary restriction. Here, we show that Pten loss is not sufficient to drive AKT activation and resistance to dietary restriction in tumors with low growth factor receptor levels. We describe a murine Pten-null Kras-driven lung cancer model that harbors both dietary restriction–resistant, higher-grade, bronchiolar tumors with high AKT activity, and dietary restriction–sensitive, lower-grade, alveolar tumors with low AKT activity. We find that this phenotype is cell autonomous and that normal bronchiolar cells express higher levels of insulin-like growth factor-I receptor (IGF-IR) and of ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5), an endoplasmic reticulum enzyme known to modulate growth factor receptor levels. Suppression of ENTPD5 is sufficient to decrease IGF-IR levels and sensitize bronchiolar tumor cells to serum in vitro and to dietary restriction in vivo. Furthermore, we find that a significant percentage of human non–small cell lung carcinomas (NSCLC) have low AKT activity despite PTEN loss. SIGNIFICANCE: Our studies point to a heterogeneity of AKT activation in the same murine Pten-null lung tissue and in human NSCLC, further underscoring the challenges of personalizing cancer therapy based solely on cancer genotype. Our findings therefore indicate that the tumor response to anticancer therapies, including dietary restriction, needs to be based on PI3K/AKT activity per se, rather than on genetic alterations in the PTEN/PI3K pathway.Howard Hughes Medical Institute (Investigator)National Institutes of Health (U.S.) (NIH grant R01 AI047389)National Institutes of Health (U.S.) (NIH grant R01 CA129105)National Institutes of Health (U.S.) (NIH grant 2-P30-CA14051)Alexander and Margaret Stewart Trust (Award)Boston Children's HospitalDavid H. Koch Institute for Integrative Cancer Research at MIT (Anna Fuller Fund fellowship

Pten

PTEN loss is considered a biomarker for activated PI3K/Akt, a pathway frequently mutated in cancer, and recently shown to confer resistance to dietary restriction (DR). Here we demonstrate that PTEN loss is not sufficient to drive Akt activation and resistance to DR in tumors with low growth factor receptor levels. We describe a murine PTEN-null Kras-driven lung cancer model that harbors both DR-resistant, higher-grade, bronchiolar tumors with high-Akt-activity, and DR-sensitive, lower-grade, alveolar tumors with low-Akt-activity. We find that this phenotype is cell-autonomous and that normal bronchiolar cells express higher levels of IGF1R and ENTPD5, an endoplasmic reticulum (ER) enzyme known to modulate growth factor receptor levels. Suppression of ENTPD5 is sufficient to decrease IGF1R levels and sensitize bronchiolar tumor cells to serum in vitro and to DR in vivo. Furthermore, we find that a significant percentage of human non-small cell lung cancer (NSCLC) have low Akt activity despite PTEN loss

The C-terminal 50 amino acid residues of dengue NS3 protein are important for NS3-NS5 interaction and viral replication

Dengue virus multifunctional proteins NS3 protease/helicase and NS5 methyltransferase/RNA-dependent RNA polymerase form part of the viral replication complex and are involved in viral RNA genome synthesis, methylation of the 5′-cap of viral genome, and polyprotein processing among other activities. Previous studies have shown that NS5 residue Lys-330 is required for interaction between NS3 and NS5. Here, we show by competitive NS3-NS5 interaction ELISA that the NS3 peptide spanning residues 566–585 disrupts NS3-NS5 interaction but not the null-peptide bearing the N570A mutation. Small angle x-ray scattering study on NS3(172–618) helicase and covalently linked NS3(172–618)-NS5(320–341) reveals a rigid and compact formation of the latter, indicating that peptide NS5(320–341) engages in specific and discrete interaction with NS3. Significantly, NS3:Asn-570 to alanine mutation introduced into an infectious DENV2 cDNA clone did not yield detectable virus by plaque assay even though intracellular double-stranded RNA was detected by immunofluorescence. Detection of increased negative-strand RNA synthesis by real time RT-PCR for the NS3:N570A mutant suggests that NS3-NS5 interaction plays an important role in the balanced synthesis of positive- and negative-strand RNA for robust viral replication. Dengue virus infection has become a global concern, and the lack of safe vaccines or antiviral treatments urgently needs to be addressed. NS3 and NS5 are highly conserved among the four serotypes, and the protein sequence around the pinpointed amino acids from the NS3 and NS5 regions are also conserved. The identification of the functionally essential interaction between the two proteins by biochemical and reverse genetics methods paves the way for rational drug design efforts to inhibit viral RNA synthesis.MOE (Min. of Education, S’pore)NMRC (Natl Medical Research Council, S’pore)MOH (Min. of Health, S’pore)Published versio

Characterization of the binding of DENV1-4 NS5 residues 865–900 to Impα.

<p><b>(A)</b> After pre-incubation of glutathione coated plate with GST-NLS fusion protein, 2-fold serially diluted of Impα (starting from 12.5 μM) was added. Bound Impα protein was detected with anti-6x His tag HRP antibody. Following development with the colorimetric substrate TMB, the reaction was stopped and absorbance at 450nm was taken. Data were fitted to one site specific binding equation using Graphpad Prism 6.0f from triplicate measurements of two independent experiments.<b>(B and C)</b> Result of binding assay performed with glutathione beads that were incubated with GST-NLS fusion protein. <b>(B)</b> 2-fold serially diluted of Impα (starting from 800 nM) was incubated with GST-NLS fusion protein in a final volume of 50 ml. After incubation, beads were sedimented and protein was eluted using GST B buffer. <b>(C)</b> Elutled proteins were visualised on SDS-PAGE and analysis was performed using one site specific binding function in Graphpad Prism 6.0f. Results are shown as the mean ± SD of duplicates from two independent experiments.</p

PISA interface analysis of DENV C-terminal NLS Impα bond forming residues.

<p>PISA interface analysis of DENV C-terminal NLS Impα bond forming residues.</p

Sequence variation and conformational flexibility of NS5 Cter18 regulates the oligomeric state, nuclear localization, viral replication.

<p><b>(A)</b> Sequence variation of residues 828–848 and 883–900 of DENV1–4 and representative flaviviruses. Arrow highlights the completely conserved Y838 and R888. The alignment was performed using Clustal Omega. The virus sequences and their GenBank accession numbers are as follows: DENV1-4 (same GenBank accession number as above), Japanese Encephalitis virus (JEV; M55506), West Nile virus (WNV; M12294), Murray Valley Encephalitis virus (MVE; AF161266), Tick-Borne Encephalitis Virus (TBEV; U27495), Yellow Fever virus (YFV; X15062) and Zika virus (ZIKV;KU497555) [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005886#ppat.1005886.ref058" target="_blank">58</a>]. <b>(B)</b> Model of the role of the C-terminal region of NS5 that is required for the formation of dimer and the initiation of <i>de novo</i> viral RNA replication and interaction with importin-α or NS3. On the left: <i>de novo</i> RNA synthesis by NS5 oligomers independent of NS3 interaction as implied by NS3 N570A mutation [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005886#ppat.1005886.ref059" target="_blank">59</a>] and non-viability of NS5 R888A as identified in this study. On the right: NS3 interaction with NS5 thumb domain in the RC will support higher plus strand RNA synthesis or NS5 Cter₁₈ interacts with host importins or other factors to be localized to nucleus or cytoplasm in a regulated manner.</p

Structure of the DENV2 and DENV3 Cter₁₈-Impα complex.

<p><b>(A and B)</b> Atomic resolution structures of DENV2 Cter₁₈-Impα complex and DENV3 Cter₁₈-Impα complex. <b>(A)</b> DENV2 Cter₁₈ (purple) and DENV3 Cter₁₈ (red), in complex with Impα (yellow) reveal they are both monopartite NLSs and bind Impα in the major binding pocket (ARMs 2–4). <b>(B)</b> The 3σ simulated annealing Fo-Fc omit map, supports this model and position of the c-terminal NLS residues. The biggest energetic contribution to the interaction is from K887 which forms a salt bridge with Impα residue D192, R888 is also critical for hydrogen binding Impα residues N188 and N228, as is R890 that forms hydrogen bonds with Impα residues N146 and Q181. <b>(C)</b> NLS binding determinants in the DENV1-4 Cter_18.</p