Causes and Consequences of Genetic Robustness and Fragility in HIV-1 Proteins

Genetic robustness describes a capacity to maintain function in the face of mutation. RNA viruses, like human immunodeficiency virus-1 (HIV-1), experience extremely high mutations rates in vivo, which contribute to the evolvability that permits HIV-1 to evade immune defenses. However, the compact nature of the HIV-1 viral genome, in which small or overlapping proteins are often multifunctional and essential, often necessitates a strong pressure to conserve functional roles, which is at opposition with immunological pressures to diversify. Under such conditions, and in consideration of the relatively rapid replication cycle of HIV-1, natural selection for robustness would be predicted to be beneficial. At the same time, robustness can come at the cost of fitness, and it is unclear whether robustness or fitness would constitute the dominant selective force in the natural setting. Within a given virus, genetic robustness is expected to vary amongst proteins, in accordance with specific function. Therefore, to more accurately examine genetic robustness within HIV-1, we selected one protein in which the competing needs to both conserve function yet diversify sequence is particularly acute. HIV-1 capsid (CA) is under strong pressure to preserve roles in viral assembly, maturation, uncoating and nuclear import, but is also under intense immunological pressure to diversify, particularly from adaptive immune responses. To evaluate the genetic robustness of HIV-1 CA, we generated a large library of random single amino acid substitution CA mutants. Surprisingly, after measuring the replicative fitness of the CA library mutants, we observed HIV-1 CA to be the most genetically fragile protein analyzed with such an approach, with 70% of random CA mutations resulting in nonviable, replicationdefective viruses (\u3c2% of WT fitness). While CA is involved in several steps of HIV-1 replication, analyses of conditionally (temperature-sensitive) and constitutively nonviable mutants indicated that the biological basis for the genetic fragility, or lack of robustness, was principally the need to organize accurate and efficient assembly of mature viral particles. Examination of the CA library mutations in naturally occurring HIV-1 subtype B populations indicated that all mutations occurring at a frequency \u3e3% in vivo had fitness levels that were \u3e40% of WT in vitro. Moreover, protective CTL epitopes were observed to preferentially occur in regions of HIV-1 CA that were even more genetically fragile than HIV-1 CA as a whole. Overall, these results suggest the extreme genetic fragility of CA may explain why immune responses to Gag, as opposed to other viral proteins, correlate with better prognosis in HIV-1 infection. To evaluate whether the fragility we observed in CA was unique to CA or a general property of HIV-1 proteins, and to more firmly establish causes and consequences of genetic robustness or fragility, we evaluated the robustness of another HIV-1 protein, integrase (IN). Again, a large library of unbiased single amino acid mutants was created. In contrast to CA, IN appeared comparatively robust, with only 35% of 156 single amino acid mutations resulting in nonviable viruses. However, when these nonviable mutants were mapped onto a model of the HIV-1 intasome, we noted a striking localization of nonviable mutants to certain IN subunit interfaces. To this end, single mutants affecting both particle production and IN expression in virions could be mapped to even more specific regions within the intasome model. Furthermore, after examining the prevalence of the IN library mutations in an in vivo cohort of subtype B IN sequences, mapping to the intasome helped explain why some in vitro library mutations with high fitness (\u3e40% of WT) never occur in vivo, and also revealed that residues that are highly variable in vivo are more likely to occur in regions in the intasome with more exposed surface area. Despite the relative robustness of HIV-IN, our analyses with an intasome model demonstrate that localized regions of fragility, namely certain subunit interfaces, exist. Understanding the genetic robustness of HIV-1 is particularly important given the continued need for novel therapeutic interventions. Not only has the evaluation of HIV-1 CA and IN robustness revealed particularly fragile regions in both proteins, which may prove ideal targets, but comparisons of their genetic robustness is suggestive of the direction future therapeutic research should take

The envelope gene of transmitted HIV-1 resists a late interferon gamma-induced block

Type I interferon (IFN) signaling engenders an antiviral state that likely plays an important role in constraining HIV-1 transmission and contributes to defining subsequent AIDS pathogenesis. Type II IFN (IFNγ) also induces an antiviral state but is often primarily considered to be an immunomodulatory cytokine. We report that IFNγ stimulation can induce an antiviral state that can be both distinct from that of type I interferon, and can potently inhibit HIV-1 in primary CD4+ T cells and a number of human cell lines. Strikingly, we find that transmitted/founder (TF) HIV-1 viruses can resist a late block that is induced by type II IFN, and the use of chimeric IFNγ- sensitive/resistant viruses indicates that interferon-resistance maps to the env gene. Simultaneously, in vitro evolution also revealed that just a single amino acid substitution in envelope can confer substantial resistance to IFN-mediated inhibition. Thus, the env gene of transmitted HIV-1 confers resistance to a late block that is phenotypically distinct from those previously described to be resisted by env, and is therefore mediated by unknown IFNγ-stimulated factor(s) in human CD4+ T cells and cell lines. This important unidentified block could play a key role in constraining HIV-1 transmission

Recurrent scoliosis one year after surgical correction

A year after anterolateral spondylodesis for progressive scoliosis, the patient showed a flexion gait pattern with recurrent deformity, due to late infection. Surgical debridement resolved all symptoms. Whereas most postoperative infections occur after posterior spondylodesis and present with back pain and mild increase of infection parameters, late infection after anterolateral approach is rare. In this case the patient did not present with the classic symptoms

Analysis and Testing of the FBA-11 Force [Balance] Accelerometer

The FBA-11 is a feedback-controlled accelerometer widely used to measure and record accelerations arising from earthquakes. It has found application both for structural response and for ground motion studies. The design intent of the FBA-11 was to provide electronic control of the natural frequency, damping, and output voltage. Included in this paper are (1) a circuit analysis yielding the complete closed-loop transfer function, and (2) the corroborative test results from shake table evaluations. The transfer function can be used to correct recorded accelerations for instrument response

A tailored hybrid BODIPY–oligothiophene donor for molecular bulk heterojunction solar cells with improved performances

Fixation of a 5-hexyl-2,2′-bithienyl unit on a conjugated BODIPY donor increases the conversion efficiency of the resulting molecular bulk heterojunction solar cells from 1.30 to 2.20%

Interferon-stimulated gene (ISG)-expression screening reveals the specific antibunyaviral activity of ISG20

Bunyaviruses pose a significant threat to human health, prosperity and food security. In response to viral infections, interferons (IFNs) upregulate the expression of hundreds of interferon stimulated genes (ISGs) whose cumulative action can potently inhibit the replication of bunyaviruses. We used a flow cytometry-based method to screen the ability of ∼500 unique ISGs from humans and rhesus macaques to inhibit the replication of Bunyamwera orthobunyavirus (BUNV), the prototype of both the Peribunyaviridae family and Bunyavirales order. Candidates possessing antibunyaviral activity were further examined using a panel of divergent bunyaviruses. Interestingly, one candidate, ISG20, exhibited potent antibunyaviral activity against most viruses examined from the Peribunyaviridae, Hantaviridae and Nairoviridae families, whereas phleboviruses (Phenuiviridae) largely escaped inhibition. Similar to other viruses known to be targeted by ISG20, the antibunyaviral activity of ISG20 is dependent upon its functional ribonuclease activity. Through use of an infectious VLP assay (based on the BUNV minigenome system), we confirmed that gene expression from all 3 viral segments is strongly inhibited by ISG20. Using in vitro evolution, we generated a substantially ISG20-resistant BUNV and mapped the determinants of ISG20 sensitivity/resistance. Taken together, we report that ISG20 is a broad and potent antibunyaviral factor yet some bunyaviruses are remarkably ISG20 resistant. Thus, ISG20 sensitivity/resistance could influence the pathogenesis of bunyaviruses, many of which are emerging viruses of clinical or veterinary significance

Duration of symptoms resulting from lumbar disc herniation: effect on treatment outcomes: analysis of the Spine Patient Outcomes Research Trial (SPORT).

BACKGROUND: The purpose of the present study was to determine if the duration of symptoms affects outcomes following the treatment of intervertebral lumbar disc herniation. METHODS: An as-treated analysis was performed on patients enrolled in the Spine Patient Outcomes Research Trial (SPORT) for the treatment of intervertebral lumbar disc herniation. Randomized and observational cohorts were combined. A comparison was made between patients who had had symptoms for six months or less (n = 927) and those who had had symptoms for more than six months (n = 265). Primary and secondary outcomes were measured at baseline and at regular follow-up intervals up to four years. The treatment effect for each outcome measure was determined at each follow-up period for the duration of symptoms for both groups. RESULTS: At all follow-up intervals, the primary outcome measures were significantly worse in patients who had had symptoms for more than six months prior to treatment, regardless of whether the treatment was operative or nonoperative. When the values at the time of the four-year follow-up were compared with the baseline values, patients in the operative treatment group who had had symptoms for six months or less had a greater increase in the bodily pain domain of the Short Form-36 (SF-36) (mean change, 48.3 compared with 41.9; p \u3c 0.001), a greater increase in the physical function domain of the SF-36 (mean change, 47.7 compared with 41.2; p \u3c 0.001), and a greater decrease in the Oswestry Disability Index score (mean change, -41.1 compared with -34.6; p \u3c 0.001) as compared with those who had had symptoms for more than six months (with higher scores indicating less severe symptoms on the SF-36 and indicating more severe symptoms on the Oswestry Disability Index). When the values at the time of the four-year follow-up were compared with the baseline values, patients in the nonoperative treatment group who had had symptoms for six months or less had a greater increase in the bodily pain domain of the SF-36 (mean change, 31.8 compared with 21.4; p \u3c 0.001), a greater increase in the physical function domain of the SF-36 (mean change, 29.5 compared with 22.6; p = 0.015), and a greater decrease in the Oswestry Disability Index score (mean change, -24.9 compared with -18.5; p = 0.006) as compared with those who had had symptoms for more than six months. Differences in treatment effect between the two groups related to the duration of symptoms were not significant. CONCLUSIONS: Increased symptom duration due to lumbar disc herniation is related to worse outcomes following both operative and nonoperative treatment. The relative increased benefit of surgery compared with nonoperative treatment was not dependent on the duration of the symptoms

Semi-conducteurs Isotropes et Stables pour Cellules solaires Organiques

Date du colloque : 03/2011</p

Identification of Interferon-Stimulated Genes with Antiretroviral Activity

SummaryInterferons (IFNs) exert their anti-viral effects by inducing the expression of hundreds of IFN-stimulated genes (ISGs). The activity of known ISGs is insufficient to account for the antiretroviral effects of IFN, suggesting that ISGs with antiretroviral activity are yet to be described. We constructed an arrayed library of ISGs from rhesus macaques and tested the ability of hundreds of individual macaque and human ISGs to inhibit early and late replication steps for 11 members of the retroviridae from various host species. These screens uncovered numerous ISGs with antiretroviral activity at both the early and late stages of virus replication. Detailed analyses of two antiretroviral ISGs indicate that indoleamine 2,3-dioxygenase 1 (IDO1) can inhibit retroviral replication by metabolite depletion while tripartite motif-56 (TRIM56) accentuates ISG induction by IFNα and inhibits the expression of late HIV-1 genes. Overall, these studies reveal numerous host proteins that mediate the antiretroviral activity of IFNs

Does the load-sharing classification predict ligamentous injury, neurological injury, and the need for surgery in patients with thoracolumbar burst fractures?: Clinical article.

OBJECT: The load-sharing score (LSS) of vertebral body comminution is predictive of results after short-segment posterior instrumentation of thoracolumbar burst fractures. Some authors have posited that an LSS \u3e 6 is predictive of neurological injury, ligamentous injury, and the need for surgical intervention. However, the authors of the present study hypothesized that the LSS does not predict ligamentous or neurological injury. METHODS: The prospectively collected spinal cord injury database from a single institution was queried for thoracolumbar burst fractures. Study inclusion criteria were acute (\u3c 24 hours) burst fractures between T-10 and L-2 with preoperative CT and MRI. Flexion-distraction injuries and pathological fractures were excluded. Four experienced spine surgeons determined the LSS and posterior ligamentous complex (PLC) integrity. Neurological status was assessed from a review of the medical records. RESULTS: Forty-four patients were included in the study. There were 4 patients for whom all observers assigned an LSS \u3e 6, recommending operative treatment. Eleven patients had LSSs ≤ 6 across all observers, suggesting that nonoperative treatment would be appropriate. There was moderate interobserver agreement (0.43) for the overall LSS and fair agreement (0.24) for an LSS \u3e 6. Correlations between the LSS and the PLC score averaged 0.18 across all observers (range -0.02 to 0.34, p value range 0.02-0.89). Correlations between the LSS and the American Spinal Injury Association motor score averaged -0.12 across all observers (range -0.25 to -0.03, p value range 0.1-0.87). Correlations describing the relationship between an LSS \u3e 6 and the treating physician\u27s decision to operate averaged 0.17 across all observers (range 0.11-0.24, p value range 0.12-0.47). CONCLUSIONS: The LSS does not uniformly correlate with the PLC injury, neurological status, or empirical clinical decision making. The LSSs of only one observer correlated significantly with PLC injury. There were no significant correlations between the LSS as determined by any observer and neurological status or clinical decision making