Tryptophan 95, an Amino Acid Residue of the Caprine Arthritis Encephalitis Virus Vif Protein Which Is Essential for Virus Replication

AbstractThe Caprine arthritis encephalitis virus (CAEV) vif gene was demonstrated to be essential for efficient virus replication. CAEV Vif deletion mutants demonstrated an attenuated replication phenotype in primary goat cell cultures and resulted in abortive infection when inoculated into goats. In this study, we determined the in vitro replication phenotype of five CAEV Vif point mutant infectious molecular clones and the ability of the corresponding in vitro translated Vif proteins to interact with the CAEV Pr55gag in the glutathione S–transferase (GST) binding assay. Here we show that (i) three of the mutants (S170E, S170G, S197G) behaved as the wild-type CAEV according to virus replication and Vif–Gag interactions; (ii) one mutant (Vif 6mut) was replication incompetent and bound weakly to GST–Gag fusion proteins; and (iii) one mutant (Vif RG) was impaired for replication while retaining its interaction properties. This mutant points out the critical importance of the CAEV Vif tryptophan residue at position 95 for efficient virus replication, defining for this lentivirus a functional domain unrelated to the Gag binding region

An Investigation of Maize at Four Sites (LA 20241, LA 38597, LA 112766, and LA 131202) in Eddy County, New Mexico

A topic of interest for many New Mexico archaeologists is the introduction and domestication of maize in the Southwest. This investigation adds to the archaeological record of when and to what extent maize was integrated into the subsistence of southeastern New Mexico prehistoric groups. Currently, the accepted date range for the introduction of maize in southeast New Mexico is 500–200 BC (Vierra 2020). Preliminary results of this investigation indicate the presence of maize in the Permian Basin of southeastern New Mexico dating to 2501 +/-125 calibrated (cal) BC; 1000 years prior to the earliest maize site recorded in the archaeological record for the area. The significance of this early date is twofold 1) the Middle Archaic date in comparison to other old maize sites in the area; and 2) the Middle Archaic date challenges the currently accepted migration patterns of maize into southeastern New Mexico. Dr. Jonathan Mabry’s 2008 study suggest that maize was introduced no later than 2100 BC in the southwest; however, Mabry states that maize use did not become common in the North American southwest until around 1400 BC (Mabry 2008). This investigation focuses on a case study of four sites, LA 20241, LA 38597, LA 112766, and LA 131202, in what is now known as Eddy County within the Permian Basin of southeastern New Mexico. I chose these sites because of my direct involvement in the data recovery field investigation and curation. I spent several weeks directing the excavation at Sites LA 112766 and LA 131202. and served as the laboratory manager for processing the artifact collections and flotation samples for all four sites. Evidence recovered from these four archaeological sites in southeast New Mexico, specifically Eddy County, suggest that maize use was low through the Archaic period and did not increase until AD 700–850 (Diehl 1996, Miller 2016, Railey 2016). This thesis demonstrates that maize was present much earlier in the archaeological record than previously reported for southeastern New Mexico. The analysis of macrobotanical, phytolith, and starch remains, and ceramics, and radiocarbon dates from cultural features at Sites LA 20241, LA 38597, LA 112766 and LA 131202 were examined to answer the question: when and to what extent was maize integrated into the subsistence of southeastern New Mexico prehistoric groups? A radiocarbon date from Feature 5, at Site LA 112766, indicates evidence of maize as early as 2501 +/-125 calibrated (cal) BC. Additionally, radiocarbon dates identified six Late Archaic features and thirteen Early Formative features that contained maize residue collectively from Sites LA 20241, LA 38597, LA 112766, and LA 131202. Lastly, Site LA 20241 had a single Late Formative feature that yielded maize residue. This thesis will focus on the signature of maize in the archaeological record of Archaic and Formative groups of southeastern New Mexico

Revised Mimivirus major capsid protein sequence reveals intron-containing gene structure and extra domain

Background: Acanthamoebae polyphaga Mimivirus (APM) is the largest known dsDNA virus. The viral particle has a nearly icosahedral structure with an internal capsid shell surrounded with a dense layer of fibrils. A Capsid protein sequence, D13L, was deduced from the APM L425 coding gene and was shown to be the most abundant protein found within the viral particle. However this protein remained poorly characterised until now. A revised protein sequence deposited in a database suggested an additional N-terminal stretch of 142 amino acids missing from the original deduced sequence. This result led us to investigate the L425 gene structure and the biochemical properties of the complete APM major Capsid protein. Results: This study describes the full length 3430 bp Capsid coding gene and characterises the 593 amino acids long corresponding Capsid protein 1. The recombinant full length protein allowed the production of a specific monoclonal antibody able to detect the Capsid protein 1 within the viral particle. This protein appeared to be post-translationnally modified by glycosylation and phosphorylation. We proposed a secondary structure prediction of APM Capsid protein 1 compared to the Capsid protein structure of Paramecium Bursaria Chlorella Virus 1, another member of the Nucleo-Cytoplasmic Large DNA virus family. Conclusion: The characterisation of the full length L425 Capsid coding gene of Acanthamoebae polyphaga Mimivirus provides new insights into the structure of the main Capsid protein. The production of a full length recombinant protein will be useful for further structural studies

Requirement of Caprine Arthritis Encephalitis VirusvifGene forin VivoReplication

AbstractReplication ofvif-caprine arthritis encephalitis virus (CAEV) is highly attenuated in primary goat synovial membrane cells and blood-derived macrophages compared to the wild-type (wt) virus. We investigated the requirement for CAEV Vif forin vivoreplication and pathogenicity in goats by intra-articular injection of either infectious proviral DNA or viral supernatants. Wild-type CAEV DNA or virus inoculation induced persistent infection resulting in severe inflammatory arthritic lesions in the joints. We were unable to detect any sign of virus replication invif-CAEV DNA inoculated goats, whilevif-CAEV virus inoculation resulted in the seroconversion of the goats. However, virus isolation and RT-PCR analyses on blood-derived macrophage cultures remained negative throughout the experiment as well as in joint or lymphoid tissues taken at necropsy. No pathologic lesions could be observed in joint tissue sections examined at necropsy. Goats inoculated with thevif-virus demonstrated no protection against a pathogenic virus challenge. These results demonstrate that CAEV Vif is absolutely required for efficientin vivovirus replication and pathogenicity and provide additional evidence that live attenuated lentiviruses have to establish a persistent infection to induce efficient protective immunity

Proceedings of the 2019 Canadian Inflammatory Myopathy Study Symposium: Clinical Trial Readiness in Myositis.

The Canadian Inflammatory Myopathy Study (CIMS) is a multicenter prospective cohort recruiting in 8 centers across Canada. One of the aims of CIMS is to conduct and participate in clinical trials in autoimmune inflammatory myopathies (AIM). Conducting clinical trials in rare diseases such as AIM presents challenges. During this symposium, experts in the field presented different solutions to successfully conduct clinical trials in AIM, including the importance of collaboration and careful trial design, as well as training and mentoring of young investigators

Reprocessability of PHB in extrusion: ATR-FTIR, tensile tests and thermal studies

Mechanical recycling of biodegradable plastics has to be encouraged, since the consumption of energy and raw materials can be reduced towards a sustainable development in plastics materials. In this study, the evolution of thermal and mechanical properties, as well as structural changes of poly(hydroxybutyrate) (PHB) up to three extrusion cycles were investigated. Results indicated a significant reduction in mechanical properties already at the second extrusion cycle, with a reduction above 50% in the third cycle. An increase in the crystallinity index was observed due to chemicrystallization process during degradation by chain scission. On the other hand, significant changes in the chemical structure or in thermal stability of PHB cannot be detected by Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analyses (TGA), respectively.National Foundation for Science and Technology Development (CNPq)Univ Fed Rio Grande Norte UFRN, Dept Mat Engn DEMat, Natal, RN, BrazilUniv Fed Sao Carlos UFSCar, Dept Mat Engn DEMa, Sao Carlos, SP, BrazilUniv Fed Paraiba UFPB, Dept Mat Engn DEMAT, Joao Pessoa, Paraiba, BrazilUniv Fed Rio Grande Norte UFRN, Dept Chem DQ, Natal, RN, BrazilUniv Fed Sao Paulo UNIFESP, Inst Sci & Technol ICT, Sao Jose Dos Campos, SP, BrazilUniv Fed Sao Paulo UNIFESP, Inst Sci & Technol ICT, Sao Jose Dos Campos, SP, BrazilWeb of Scienc

Ultrastructural Characterization of the Giant Volcano-like Virus Factory of Acanthamoeba polyphaga Mimivirus

Acanthamoeba polyphaga Mimivirus is a giant double-stranded DNA virus defining a new genus, the Mimiviridae, among the Nucleo-Cytoplasmic Large DNA Viruses (NCLDV). We used utrastructural studies to shed light on the different steps of the Mimivirus replication cycle: entry via phagocytosis, release of viral DNA into the cell cytoplasm through fusion of viral and vacuolar membranes, and finally viral morphogenesis in an extraordinary giant cytoplasmic virus factory (VF). Fluorescent staining of the AT-rich Mimivirus DNA showed that it enters the host nucleus prior to the generation of a cytoplasmic independent replication centre that forms the core of the VF. Assembly and filling of viral capsids were observed within the replication centre, before release into the cell cytoplasm where progeny virions accumulated. 3D reconstruction from fluorescent and differential contrast interference images revealed the VF emerging from the cell surface as a volcano-like structure. Its size dramatically grew during the 24 h infectious lytic cycle. Our results showed that Mimivirus replication is an extremely efficient process that results from a rapid takeover of cellular machinery, and takes place in a unique and autonomous giant assembly centre, leading to the release of a large number of complex virions through amoebal lysis

Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5

The membrane attack complex (MAC or C5b-9) is an important effector of the immune system to kill invading microbes. MAC formation is initiated when complement enzymes on the bacterial surface convert complement component C5 into C5b. Although the MAC is a membrane-inserted complex, soluble forms of MAC (sMAC), or terminal complement complex (TCC), are often detected in sera of patients suffering from infections. Consequently, sMAC has been proposed as a biomarker, but it remains unclear when and how it is formed during infections. Here, we studied mechanisms of MAC formation on different Gram-negative and Gram-positive bacteria and found that sMAC is primarily formed in human serum by bacteria resistant to MAC-dependent killing. Surprisingly, C5 was converted into C5b more potently by MAC-resistant compared to MAC-sensitive Escherichia coli strains. In addition, we found that MAC precursors are released from the surface of MAC-resistant bacteria during MAC assembly. Although release of MAC precursors from bacteria induced lysis of bystander human erythrocytes, serum regulators vitronectin (Vn) and clusterin (Clu) can prevent this. Combining size exclusion chromatography with mass spectrom-etry profiling, we show that sMAC released from bacteria in serum is a heterogeneous mixture of complexes composed of C5b-8, up to three copies of C9 and multiple copies of Vn and Clu. Alto-gether, our data provide molecular insight into how sMAC is generated during bacterial infections. This fundamental knowledge could form the basis for exploring the use of sMAC as biomarker

Interventions targeting glucocorticoid-Krüppel-like factor 15-branched-chain amino acid signaling improve disease phenotypes in spinal muscular atrophy mice

The circadian glucocorticoid-Krüppel-like factor 15-branched-chain amino acid (GC-KLF15-BCAA) signaling pathway is a key regulatory axis in muscle, whose imbalance has wide-reaching effects on metabolic homeostasis. Spinal muscular atrophy (SMA) is a neuromuscular disorder also characterized by intrinsic muscle pathologies, metabolic abnormalities and disrupted sleep patterns, which can influence or be influenced by circadian regulatory networks that control behavioral and metabolic rhythms. We therefore set out to investigate the contribution of the GC-KLF15-BCAA pathway in SMA pathophysiology of Taiwanese Smn−/−;SMN2 and Smn2B/− mouse models. We thus uncover substantial dysregulation of GC-KLF15-BCAA diurnal rhythmicity in serum, skeletal muscle and metabolic tissues of SMA mice. Importantly, modulating the components of the GC-KLF15-BCAA pathway via pharmacological (prednisolone), genetic (muscle-specific Klf15 overexpression) and dietary (BCAA supplementation) interventions significantly improves disease phenotypes in SMA mice. Our study highlights the GC-KLF15-BCAA pathway as a contributor to SMA pathogenesis and provides several treatment avenues to alleviate peripheral manifestations of the disease. The therapeutic potential of targeting metabolic perturbations by diet and commercially available drugs could have a broader implementation across other neuromuscular and metabolic disorders characterized by altered GC-KLF15-BCAA signaling

Determinants of participation in a web-based health risk assessment and consequences for health promotion programs

Background: The health risk assessment (HRA) is a type of health promotion program frequently offered at the workplace. Insight into the underlying determinants of participation is needed to evaluate and implement these interventions. Objective: To analyze whether individual characteristics including demographics, health behavior, self-rated health, and work-related factors are associated with participation and nonparticipation in a Web-based HRA. Methods: Determinants of participation and nonparticipation were investigated in a cross-sectional study among individuals employed at five Dutch organizations. Multivariate logistic regression was performed to identify determinants of participation and nonparticipation in the HRA after controlling for organization and all other variables. Results: Of the 8431 employees who were invited, 31.9% (2686/8431) enrolled in the HRA. The online questionnaire was completed by 27.2% (1564/5745) of the nonparticipants. Determinants of participation were some periods of stress at home or work in the preceding year (OR 1.62, 95% CI 1.08-2.42), a decreasing number of weekdays on which at least 30 minutes were spent on moderate to vigorous physical activity (ORdayPA0.84, 95% CI 0.79-0.90), and increasing alcohol consumption. Determinants of nonparticipation were less-than-positive self-rated health (poor/very poor vs very good, OR 0.25, 95% CI 0.08-0.81) and tobacco use (at least weekly vs none, OR 0.65, 95% CI 0.46-0.90). Conclusions: This study showed that with regard to isolated health behaviors (insufficient physical activity, excess alcohol consumption, and stress), those who could benefit most from the HRA were more likely to participate. However, tobacco users and those who rate