Respiratory synctial virus interactions with host-cell RNA-processing structures and proteins

Thesis (Ph.D.)--Boston UniversityRespiratory syncytial virus (RSV) is a negative-strand RNA virus that causes significant pneumonia-related morbidity and mortality worldwide. There are currently neither vaccines nor effective therapies for RSV. As with other viruses, RSV mRNAs are translated using host-cell machinery, rendering the virus subject to cellular factors that regulate mRNA homeostasis. Stress granules (SGs) and processing bodies (p-bodies) are inter-dependent, stress-response cytoplasmic structures involved in mRNA triage and degradation, respectively. We hypothesized that RSV has evolved to manipulate cellular stress responses in order to facilitate optimal virus propagation. While wild-type (wt) RSV induced SGs in approximately 1% of infected cells, a mutant version of RSV whose Tr region was replaced with an inverted LeC sequence (LeC virus) induced SG formation in approximately 50% to 70% of infected cells. A 12U to A substitution relative to the 5' end of the LeC virus abrogated SG induction. Mixed-infection studies showed that wt RSV was able to prevent LeC-mediated SG induction. Unlike Sendai virus, RSV-mediated prevention of SG formation was independent of SG-associated t-cell intracellular antigen related (TIAR) protein. RSV infection altered neither the number nor distribution of p-bodies; however, p-body-associated decapping protein 1 (dcp1) was phosphorylated throughout RSV infection via the extracellular signal-regulated kinase (ERK) 1/2 pathway. RSV-mediated dcp1 phosphorylation was limited to serine 315, serine 319, and threonine 321. Dcp1 phosphorylation occurred in response to some, but not all, environmental stresses, and dcp1 was not phosphorylated during infection with HIV-1, measles, mumps, or canine distemper virus. Overexpression of dcp1 significantly attenuated RSV cytopathic effects, and preliminary data suggested that dcp1 phosphorylation regulated RSV-induced interleukin-8 production. Finally, an antibody toward cellular SG- and p-body-associated, RNA-binding protein p54 was able to recognize a subset of RSV nucleoprotein (N). p54 and RSV N contain a similar amino acid sequence motif, suggesting that they may have similar or competing activities that are important during RSV replication. Taken together, our results demonstrate that RSV can manipulate cellular RNA-processing structures and proteins to facilitate viral propagation

UPAYA GURU DALAM MENINGKATKAN HASIL BELAJAR SISWA MATA PELAJARAN IPA PADA POKOK BAHASAN PERUBAHANWUJUD BENDA DENGAN MENGGUNAKAN MODEL PEMBELAJARAN INKUIRI (PTK di Kelas IVMI Al Washliyah Perbutulan Kecamatan Sumber Kabupaten Cirebon)

Yuyun Yuniah : Siswa kelas IV MI Al Washliyah Perbutulan Kecamatan Sumber Kabupaten Cirebon, pada hasil nilai ulangan harian IPA pada pokok bahasan perubahan wujud benda, masih banyak siswa yang tidak tuntas belajar, 21 dari 30 siswa nilainya masih di bawah KKM (Kriteria Ketuntasan Minimal). Nilai KKM IPA kelas IV tahun ajaran 2012/2013 adalah 70. Siswa kelas IV juga memiliki karakteristik aktivitas siswa yang pasif, yaitu siswa kurang komunikatif dengan guru dan teman sekelasnya. Selama pembelajaran IPA, keterlibatan siswa jarang di libatkan dalam praktikum. Untuk mengatasi permasalahan ini, maka diperlukan suatu model pembelajaran yang banyak melibatkan siswa sehingga dapat meningkatkan aktivitas hasil dan ketuntasan belajar siswa. Salah satu model yang digunakan adalah model inkuiri pada pembelajaran IPA. Pengajaran berdasarkan inkuiri adalah suatu strategi yang berpusat pada siswa di mana kelompok siswa inkuiri kedalam suatu isu atau mencari jawabanjawaban terhadap isi pertanyaan melalui suatu prosedur yang digariskan secara jelas dan struktural kelompok. Tujuan dari penelitian ini adalah (1) Untuk mengkaji keberhasilan penerapan model inkuiri, pada proses pembelajaran IPA pada pokok bahasan perubahan wujud benda di kelas IV, MI Al Washliyah Perbutulan. (2) Untuk mengkaji hasil belajar siswa kelas IV MI Al Washliyah Perbutulan pada pelajaran Ilmu Pengetahuan Alam pada pokok bahasan perubahan wujud benda. (3) Seberapa besar pengaruh penggunaan inkuiri, dengan hasil dan ketuntasan belajar siswa pada pelajaran Ilmu Pengetahuan Alam pada pokok bahasan perubahan wujud benda di kelas IV MI Al Washliyah Perbutulan. Peneliti ini dilakukan di kelas IV MI Al Washliyah Perbutulan Kecamatan Sumber Kabupaten Cirebon, semester ganjil tahun 2012/2013 yang berjumlah 30 siswa dengan menggunakan desain penelitian tindakan kelas dalam dua siklus. Tiap-tiap siklus terdiri dari empat tahap yaitu perencanaan, pelaksanaan tindakan, pengamatan dan refleksi. Teknik pengumpulan data dilakukan dengan cara observasi, wawancara dan penilaian hasil belajar. Berdasarkan hasil penelitian diperoleh nilai rata-rata hasil belajar siswa pada siklus I adalah 61,33 dan siklus II adalah 79. Secara umum dapat disimpulkan bahwa pembelajaran inkuiri dapat meningkatkan aktivitas siswa

An Economic Assessment of Patent Settlements in the Pharmaceutical Industry

This article demonstrates that in recent years, patent settlements between branded and generic manufacturers involving reverse payments from branded manufacturers to generic manufacturers have received close antitrust scrutiny, driven by concerns that such settlements harm consumers by delaying the entry of lower-priced generic drugs. The authors note that such settlements will be a focus of the Obama Administration\u27s antitrust enforcement policy, yet there is a growing consensus among the courts that such settlements are anticompetitive only under narrow sets of circumstances. In this article, the authors present an analytical framework for evaluating the competitive effects of patent settlements, including those involving reverse payments, and demonstrate that these settlements can benefit consumers. Thus, the authors conclude that while continued scrutiny of such settlements is important, broad brush treatments are inappropriate and only a more individualized evaluation can correctly determine the competitive effects of a particular settlement agreement

Guidelines for the treatment of bacterial vaginosis: focus on tinidazole

Bacterial vaginosis (BV) is a complex vaginal infection most commonly associated with women of child-bearing age. Risk factors for BV are numerous. There are currently multiple clinical and laboratory tests for diagnosis of BV, including the most commonly used diagnostic methods: Amsel’s criteria or Nugent’s Gram stain scale. The mainstay of BV therapy is metronidazole, but tinidazole as well as a few other agents have also been used successfully. Tinidazole is the second nitroimidazole antiprotozoal agent and a structural derivative of metronidazole. With a favorable pharmacokinetic profile and reduced side effects, tinidazole is an alternative agent for BV treatment. There are minimal head-to-head comparative data to establish tinidazole’s superiority to metronidazole or other therapeutic agents. Available data suggest tinidazole has a role in special populations particularly for refractory or relapsing BV

Хохотнул, схитрил: The Relationship between Semelfactives Formed with -NU- and S- in Russian

To appear in Russian LinguisticsWe know relatively little about the relationship between verbs with the suffix -nu-, such as крикнуть ‘shout once’, and verbs with the prefix s-, such as сглупить ‘do something stupid’ and сходить ‘walk someplace and back once’. In her Cluster Model of Russian aspect Janda (2007) claims that there is a single group of such Perfectives, namely the Single Act Perfectives. In order to evaluate this claim, we have gathered a database that includes both types of Single Act Perfectives and undertaken a statistical analysis. On the basis of this analysis we show that the distribution of the -nu- and s- morphemes is very much dependent on the morphological class of the verb. In addition, we have traced the historical development of the meaning of semelfactivity for the prefix s- and our study shows that this development comports well with the development of actional meanings for prefixes in Russian in the sixteenth and seventeenth centuries. This article presents new data concerning semelfactive Perfectives in Russian, along with an analysis of this data. The article raises questions about the historical development of aspect in Russian and about allomorphy in general

Hook3 is a scaffold for the opposite-polarity microtubule-based motors cytoplasmic dynein-1 and KIF1C.

The unidirectional and opposite-polarity microtubule-based motors, dynein and kinesin, drive long-distance intracellular cargo transport. Cellular observations suggest that opposite-polarity motors may be coupled. We recently identified an interaction between the cytoplasmic dynein-1 activating adaptor Hook3 and the kinesin-3 KIF1C. Here, using in vitro reconstitutions with purified components, we show that KIF1C and dynein/dynactin can exist in a complex scaffolded by Hook3. Full-length Hook3 binds to and activates dynein/dynactin motility. Hook3 also binds to a short region in the "tail" of KIF1C, but unlike dynein/dynactin, this interaction does not activate KIF1C. Hook3 scaffolding allows dynein to transport KIF1C toward the microtubule minus end, and KIF1C to transport dynein toward the microtubule plus end. In cells, KIF1C can recruit Hook3 to the cell periphery, although the cellular role of the complex containing both motors remains unknown. We propose that Hook3's ability to scaffold dynein/dynactin and KIF1C may regulate bidirectional motility, promote motor recycling, or sequester the pool of available dynein/dynactin activating adaptors

Disrupted CXCR2 Signaling in Oligodendroglia Lineage Cells Enhances Myelin Repair in a Viral Model of Multiple Sclerosis.

CXCR2 is a chemokine receptor expressed on oligodendroglia that has been implicated in the pathogenesis of neuroinflammatory demyelinating diseases as well as enhancement of the migration, proliferation, and myelin production by oligodendroglia. Using an inducible proteolipid protein (Plp) promoter-driven Cre-loxP recombination system, we were able to assess how timed ablation of Cxcr2 in oligodendroglia affected disease following intracranial infection with the neurotropic JHM strain of mouse hepatitis virus (JHMV). Generation of Plp-Cre-ER(T)::Cxcr2flox/flox transgenic mice (termed Cxcr2-CKO mice) allows for Cxcr2 to be silenced in oligodendrocytes in adult mice following treatment with tamoxifen. Ablation of oligodendroglia Cxcr2 did not influence clinical severity in response to intracranial infection with JHMV. Infiltration of activated T cells or myeloid cells into the central nervous system (CNS) was not affected, nor was the ability to control viral infection. In addition, the severity of demyelination was similar between tamoxifen-treated mice and vehicle-treated controls. Notably, deletion of Cxcr2 resulted in increased remyelination, as assessed by g-ratio (the ratio of the inner axonal diameter to the total outer fiber diameter) calculation, compared to that in vehicle-treated control mice. Collectively, our findings argue that CXCR2 signaling in oligodendroglia is dispensable with regard to contributing to neuroinflammation, but its deletion enhances remyelination in a preclinical model of the human demyelinating disease multiple sclerosis (MS).IMPORTANCE Signaling through the chemokine receptor CXCR2 in oligodendroglia is important for developmental myelination in rodents, while chemical inhibition or nonspecific genetic deletion of CXCR2 appears to augment myelin repair in animal models of the human demyelinating disease multiple sclerosis (MS). To better understand the biology of CXCR2 signaling on oligodendroglia, we generated transgenic mice in which Cxcr2 is selectively ablated in oligodendroglia upon treatment with tamoxifen. Using a viral model of neuroinflammation and demyelination, we demonstrate that genetic silencing of CXCR2 on oligodendroglia did not affect clinical disease, neuroinflammation, or demyelination, yet there was increased remyelination. These findings support and extend previous findings suggesting that targeting CXCR2 may offer a therapeutic avenue for enhancing remyelination in patients with demyelinating diseases

Biofeedback as an intervention for persistent post-concussive symptoms: A randomized feasibility trial

Background Case reports indicate that low-resolution electromagnetic tomography neurofeedback and heart rate variability biofeedback may improve physiological functioning in individuals with persistent post-concussive symptoms. However, it is unclear whether larger-scale studies are feasible. Purpose To evaluate the feasibility of a combined low-resolution electromagnetic tomography neurofeedback and heart rate variability biofeedback intervention for individuals with persistent post-concussive symptoms. Methods Individuals with persistent post-concussive symptoms were randomized into intervention and control groups, and their baseline and post-test assessments were compared to a healthy control group. Outcomes included self-report questionnaires, resting electroencephalograph and electrocardiograph recordings, and a driving simulation task. Participants in the intervention group completed three 20 min low-resolution electromagnetic tomography neurofeedback sessions per week and at-home heart rate variability biofeedback training every morning and night for 8 weeks. Feasibility was evaluated according to recruitment capability and sample characteristics, data collection procedures, suitability of the intervention and study procedures, management and implementation of the study intervention, and preliminary participant responses to the intervention. Results Thirty-three individuals were recruited and 24 completed this study (seven intervention participants, nine persistent post-concussive symptoms control participants, and eight healthy control participants). One-quarter of participants (four intervention participants and three persistent post-concussive symptoms control participants) experienced simulator sickness during the driving simulator task and had to withdraw from the study. Intervention participants had an 88% and 86% compliance rate for the low-resolution electromagnetic tomography neurofeedback and heart rate variability biofeedback sessions, respectively. Low-resolution electromagnetic tomography neurofeedback sessions took approximately 1 h to complete per participant. Preliminary analysis indicated that the intervention reduced electroencephalograph z-score deviation with a very large effect size (d = 1.36) compared to the other study groups. Conclusions Pilot studies evaluating the efficacy of low-resolution electromagnetic tomography neurofeedback and heart rate variability biofeedback should be performed to confirm these preliminary findings. However, the protocol should be modified to reduce participant fatigue and withdrawal. This trial was registered with Clinicialtrials.gov (NCT03338036; https://clinicaltrials.gov/ct2/show/NCT03338036?term=03338036&draw=2&rank=1)

MicroRNA-155 enhances T cell trafficking and antiviral effector function in a model of coronavirus-induced neurologic disease

BackgroundMicroRNAs (miRNAs) are noncoding RNAs that modulate cellular gene expression, primarily at the post-transcriptional level. We sought to examine the functional role of miR-155 in a model of viral-induced neuroinflammation.MethodsAcute encephalomyelitis and immune-mediated demyelination were induced by intracranial injection with the neurotropic JHM strain of mouse hepatitis virus (JHMV) into C57BL/6 miR-155 (+/+) wildtype (WT) mice or miR-155 (-/-) mice. Morbidity and mortality, viral load and immune cell accumulation in the CNS, and spinal cord demyelination were assessed at defined points post-infection. T cells harvested from infected mice were used to examine cytolytic activity, cytokine activity, and expression of certain chemokine receptors. To determine the impact of miR-155 on trafficking, T cells from infected WT or miR-155 (-/-) mice were adoptively transferred into RAG1 (-/-) mice, and T cell accumulation into the CNS was assessed using flow cytometry. Statistical significance was determined using the Mantel-Cox log-rank test or Student's T tests.ResultsCompared to WT mice, JHMV-infected miR-155 (-/-) mice developed exacerbated disease concomitant with increased morbidity/mortality and an inability to control viral replication within the CNS. In corroboration with increased susceptibility to disease, miR-155 (-/-) mice had diminished CD8(+) T cell responses in terms of numbers, cytolytic activity, IFN-γ secretion, and homing to the CNS that corresponded with reduced expression of the chemokine receptor CXCR3. Both IFN-γ secretion and trafficking were impaired in miR-155 (-/-) , virus-specific CD4(+) T cells; however, expression of the chemokine homing receptors analyzed on CD4(+) cells was not affected. Except for very early during infection, there were not significant differences in macrophage infiltration into the CNS between WT and miR-155 (-/-) JHMV-infected mice, and the severity of demyelination was similar at 14 days p.i. between WT and miR-155 (-/-) JHMV-infected mice.ConclusionsThese findings support a novel role for miR-155 in host defense in a model of viral-induced encephalomyelitis. Specifically, miR-155 enhances antiviral T cell responses including cytokine secretion, cytolytic activity, and homing to the CNS in response to viral infection. Further, miR-155 can play either a host-protective or host-damaging role during neuroinflammation depending on the disease trigger