Miracle-dz/UniFAR-FTUs huggingface dataset:
Pre-clinical development of cell culture (Vero)-derived H5N1 pandemic vaccines

Abstract The rapid spread of avian influenza (H5N1) and its transmission to humans has raised the possibility of an imminent pandemic and concerns over the ability of standard influenza vaccine production methods to supply sufficient amounts of an effective vaccine. We report here on a robust and flexible strategy which uses wild-type virus grown in a continuous cell culture (Vero) system to produce an inactivated whole virus vaccine. Candidate vaccines based on clade 1 and clade 2 influenza H5N1 strains, produced at a variety of manufacturing scales, were demonstrated to be highly immunogenic in animal models without the need for adjuvant. The vaccines induce cross-neutralising antibodies and are protective in a mouse challenge model not only against the homologous virus but against other H5N1 strains, including those from other clades. These data indicate that cell culture-grown, whole virus vaccines, based on the wild-type virus, allow the rapid high-yield production of a candidate pandemic vaccine.
Mathematical modeling of electromagnetic transmission in flat heterogeneous guided systems

Mathematical simulation of electromagnetic wave propagation in the flat isotropic heterogeneous linear guided systems is considered in the presence of expofunctional influences. Main types of boundary problems are proposed as analytical models of the aforesaid electrodynamic phenomena in excited media. All given statements have different boundary and initial conditions but are based on one and the same general wave equation regarding all unknown scalar components of electromagnetic field vector intensities. In its turn, originally, this wave PDE (partial differential equation) is generated by the specific case of differential Maxwell system. Equivalence of both objects in class of non generalized functions is proved by criterion whose additional solvability conditions allow formulating respective electrodynamic boundary problems correctly.
Debt and Foregone Medical Care

Most American households carry debt, yet we have little understanding of how debt influences health behavior, especially health care seeking. We examined associations between foregone medical care and debt using a population-based sample of 914 southeastern Michigan residents surveyed in the wake of the late-2000s recession. Overall debt and ratios of debt to income and debt to assets were positively associated with foregoing medical or dental care in the past 12 months, even after adjusting for the poorer socioeconomic and health characteristics of those foregoing care and for respondents’ household incomes and net worth. These overall associations were driven largely by credit card and medical debt, while housing debt and automobile and student loans were not associated with foregoing care. These results suggest that debt is an understudied aspect of health stratification.
Omentin Val109Asp polymorphism and risk of coronary artery disease

Background Omentin is an adipocytokine with antiinflammatory properties. It has been reported to be involved in atherosclerosis and coronary artery disease. We aimed to investigate the association of omentin Val109Asp polymorphism with coronary artery disease in an Iranian population. Methods For a case-control study, 400 individuals were recruited: 200 with coronary artery disease and 200 healthy controls. Patients with coronary artery disease were diagnosed by angiography as having at least one main coronary artery with more than 50% stenosis. Genotyping of Val109Asp was carried out using a polymerase chain reaction DNA-restriction fragment length polymorphism technique. Results There was no association between Val109Asp polymorphism and the risk of coronary artery disease in our study population (p = 0.20). However, when subgroup analysis was performed according to sex, there was a significant difference in the distribution of alleles between groups for men (p = 0.031, odds ratio = 0.57, 95% confidence interval: 0.35–0.95) but not for women (p = 0.88, odds ratio = 1.03, 95% confidence interval: 0.66–1.61). Conclusions Our results indicate that the Asp allele of Val109Asp (T allele of rs2274907) is more frequent among men with coronary artery disease than healthy men, so it is possibly a risk factor for coronary artery disease in men only. The difference in association between men and women may be due to the different distribution and metabolism of adipose tissue in men and women. More studies with larger sample sizes and in different populations are required to validate our study results.
Cell fate determination in cisplatin resistance and chemosensitization

Understanding the determination of cell fate choices after cancer treatment will shed new light on cancer resistance. In this study, we quantitatively analyzed the individual cell fate choice in resistant UM-SCC-38 head and neck cancer cells exposed to cisplatin. Our study revealed a highly heterogeneous pattern of cell fate choices in UM-SCC-38 cells, in comparison to that of the control, non-tumorigenic keratinocyte HaCaT cells. In both UM-SCC-38 and HaCaT cell lines, the majority of cell death occurred during the immediate interphase without mitotic entry, whereas significant portions of UM-SCC-38 cells survived the treatment via either checkpoint arrest or checkpoint slippage. Interestingly, checkpoint slippage occurred predominantly in cells treated in late S and G2 phases, and cells in M-phase were hypersensitive to cisplatin. Moreover, although the cisplatin-resistant progression of mitosis exhibited no delay in general, prolonged mitosis was correlated with the induction of cell death in mitosis. The finding thus suggested a combinatorial treatment using cisplatin and an agent that blocks mitotic exit. Consistently, we showed a strong synergy between cisplatin and the proteasome inhibitor Mg132. Finally, targeting the DNA damage checkpoint using inhibitors of ATR, but not ATM, effectively sensitized UM-SCC-38 to cisplatin treatment. Surprisingly, checkpoint targeting eliminated both checkpoint arrest and checkpoint slippage, and augmented the induction of cell death in interphase without mitotic entry. Taken together, our study, by profiling cell fate determination after cisplatin treatment, reveals new insights into chemoresistance and suggests combinatorial strategies that potentially overcome cancer resistance.
Activation of Apoptosis Signal-Regulating Kinase 1 (ASK1) by Tumor Necrosis Factor Receptor-Associated Factor 2 Requires Prior Dissociation of the ASK1 Inhibitor Thioredoxin

ABSTRACT The stress-activated protein kinases (SAPKs, also called c-Jun NH2-terminal kinases) and the p38s, two mitogen-activated protein kinase (MAPK) subgroups activated by cytokines of the tumor necrosis factor (TNF) family, are pivotal to the de novo gene expression elicited as part of the inflammatory response. Apoptosis signal-regulating kinase 1 (ASK1) is a MAPK kinase kinase (MAP3K) that activates both the SAPKs and p38s in vivo. Here we show that TNF receptor (TNFR) associated factor 2 (TRAF2), an adapter protein that couples TNFRs to the SAPKs and p38s, can activate ASK1 in vivo and can interact in vivo with the amino- and carboxyl-terminal noncatalytic domains of the ASK1 polypeptide. Expression of the amino-terminal noncatalytic domain of ASK1 can inhibit TNF and TRAF2 activation of SAPK. TNF can stimulate the production of reactive oxygen species (ROS), and the redox-sensing enzyme thioredoxin (Trx) is an endogenous inhibitor of ASK1. We also show that expression of TRAF2 fosters the production of ROS in transfected cells. We demonstrate that Trx significantly inhibits TRAF2 activation of SAPK and blocks the ASK1-TRAF2 interaction in a reaction reversed by oxidants. Finally, the mechanism of ASK1 activation involves, in part, homo-oligomerization. We show that expression of ASK1 with TRAF2 enhances in vivo ASK1 homo-oligomerization in a manner dependent, in part, upon the TRAF2 RING effector domain and the generation of ROS. Thus, activation of ASK1 by TNF requires the ROS-mediated dissociation of Trx possibly followed by the binding of TRAF2 and consequent ASK1 homo-oligomerization.
Coordination Deficits Induced in Young Adult Mice Treated with Methylmercury

Male and female C57BL/6J mice starting at postnatal (P) day 34 were exposed orally to five divided doses totaling 1.0 or 5.0 mg/kg of methylmercury (MeHg; given as methylmercuric chloride) or sterile deionized water in moistened rodent chow. After a 5-day waiting period, control and MeHg-treated mice were subjected to a standard battery of behavior tests for balance and motor coordination. Latency to falling on the accelerating rota-rod was significantly decreased in 5.0 mg/kg MeHg-exposed mice when compared to control mice. In the open field, horizontal exploration with respect to total distance traveled during the first 5 min on the first test day was significantly reduced in 1.0 mg/kg MeHg-exposed mice when compared to control mice. Rearing activity was not affected by MeHg treatment. In the footprint analysis, angle of foot placement measured in 1.0 mg/kg MeHg-treated mice was significantly greater compared to control mice. Base stance and stride length were unaffected by MeHg treatment. On the vertical pole test, 10 mice from each treatment group fell off the pole during the time the pole was shifted from a horizontal position to a vertical position, whereas none of the control mice fell. These results indicate that short-term, low to moderate doses of MeHg in young adult mice can be detrimental to motor coordination and balance.
Characterization of scFv-Ig constructs generated from the anti-CD20 mAb 1F5 using linker peptides of varying lengths.

The heavy (VH) and light (VL) chain variable regions of the murine anti-human CD20 mAb 1F5 were cloned, and four single-chain Ab (scFv) molecules were constructed using linker peptides of variable lengths to join the VH and VL domains. Three constructs were engineered using linker peptides of 15, 10, and 5 aa residues consisting of (GGGGS)3, (GGGGS)2, and (GGGGS)1 sequences, respectively, whereas the fourth was prepared by joining the VH and VL domains directly. Each construct was fused to a derivative of human IgG1 (hinge plus CH2 plus CH3) to facilitate purification using staphylococcal protein A. The aggregation and CD20 binding properties of these four 1F5 scFv-Ig derivatives produced were investigated. Both size-exclusion HPLC column analysis and Western blots of proteins subjected to nonreducing SDS-PAGE suggested that all four 1F5 scFv-Ig were monomeric with m.w. of approximately 55 kDa. The CD20 binding properties of the four 1F5 scFv-Ig were studied by ELISA and flow cytometry. The 1F5 scFv-Ig with the 5-aa linker (GS1) demonstrated significantly superior binding to CD20-expressing target cells, compared with the other scFv-Ig constructs. Scatchard analysis of the radiolabeled monovalent GS1 scFv-Ig revealed a binding avidity of 1.35 x 108 M-1 compared with an avidity of 7.56 x 108 M-1 for the native bivalent 1F5 Ab. These findings suggest that the GS1 scFv-Ig with a short linker peptide of approximately 5 aa is the best of the engineered constructs for future studies.
Commodities’ Common Factor. An Empirical Assessment of the Markets’ Drivers

Using a generalized dynamic factor model, we identify a latent common factor in a broad sample of thirty-one commodity futures’ returns between 1996 and 2015. An investigation of sub-periods reveals an increasing correlation between the common factor and changes in gold and oil prices during the financial crisis. We also consider whether the common factors of commodity subsectors give an advantage to the pricing of commodity futures’ returns. In the cross-section of individual futures’ returns we suggest that two- or three-factor models that include energy's or agriculture's common factors can explain commodity returns. Thus, our results indicate an increasing homogeneity of the commodity markets in recent years.
Matrix Protein and Another Viral Component Contribute to Induction of Apoptosis in Cells Infected with Vesicular Stomatitis Virus

ABSTRACT The induction of apoptosis in host cells is a prominent cytopathic effect of vesicular stomatitis virus (VSV) infection. The viral matrix (M) protein is responsible for several important cytopathic effects, including the inhibition of host gene expression and the induction of cell rounding in VSV-infected cells. This raises the question of whether M protein is also involved in the induction of apoptosis. HeLa or BHK cells were transfected with M mRNA to determine whether M protein induces apoptosis when expressed in the absence of other viral components. Expression of M protein induced apoptotic morphological changes and activated caspase-3 in both cell types, indicating that M protein induces apoptosis in the absence of other viral components. An M protein containing a point mutation that renders it defective in the inhibition of host gene expression (M51R mutation) activated little, if any, caspase-3, while a deletion mutant lacking amino acids 4 to 21 that is defective in the virus assembly function but fully functional in the inhibition of host gene expression was as effective as wild-type (wt) M protein in activating caspase-3. To determine whether M protein influences the induction of apoptosis in the context of a virus infection, the M51R M protein mutation was incorporated onto a wt background by using a recombinant infectious cDNA clone (rM51R-M virus). The timing of the induction of apoptosis by rM51R-M virus was compared to that by the corresponding recombinant wt (rwt) virus and to that by tsO82 virus, the mutant virus in which the M51R mutation was originally identified. In HeLa cells, rwt virus induced apoptosis faster than did rM51R-M virus, demonstrating a role for M protein in the induction of apoptosis. In contrast to the results obtained with HeLa cells, rwt virus induced apoptosis more slowly than did rM51R-M virus in BHK cells. This indicates that a viral component other than M protein contributes to induction of apoptosis in BHK cells and that wt M protein acts to delay induction of apoptosis by the other viral component. tsO82 virus induced apoptosis more rapidly than did rM51R-M virus in both HeLa and BHK cells. These two viruses contain the same point mutation in their M proteins, suggesting that sequence differences in genes other than that for M protein affect their rates of induction of apoptosis.
Cardiac Sympathovagal Balance and Peripheral Sympathetic Vasoconstriction: Epidural Versus General Anesthesia

Both epidural and general anesthetics alter autonomic balance. However, the relative differences between epidural (EA) and general anesthetics (GA) with regard to cardiac and peripheral sympathovagal balance have not been described. Twenty consecutive patients scheduled for radical retropubic prostatectomy were randomized to receive EA (n = 10) or GA (n = 10). Power spectral analysis was performed on the electrocardiographic recordings, with the ratio of low (0.05–0.125 Hz)/high (0.125–0.5 Hz) frequency power used as an index of cardiac sympathovagal balance. The forearm minus fingertip skin-surface temperature gradient (>4°C) was used as an indicator of sympathetically mediated peripheral vasoconstriction. Patients in the EA group demonstrated a significantly greater low/high frequency power ratio and a more frequent incidence of peripheral vasoconstriction than the GA group during the intraoperative period. During the postoperative period, the GA group demonstrated an increase in the low/high ratio and the incidence of vasoconstriction relative to the intraoperative period. Intraoperatively, upper body vasoconstriction appears to be accompanied by a significant shift in cardiac sympathovagal balance toward sympathetic predominance with EA relative to GA. Postoperatively, GA is associated with a shift in the sympathovagal balance toward sympathetic predominance. Further research is required to determine whether this results in cardiovascular compromise in the high-risk patient.
Genotyping of Hepatitis E Virus from Vietnam

To identify the genotype of Vietnamese isolates of human hepatitis E virus (HEV), phylogenetic analysis was performed for the open reading frame (ORF) 1 and ORF2 nucleotide sequences of the viral genome. HEV was detected by RT-PCR in 9 out of 141 sera collected from patients with a diagnosis of acute sporadic hepatitis in Hanoi, Vietnam. All of them had sequences related most closely to genotype 4. In addition, the Vietnamese isolate had a single nucleotide insertion in the ORF 3 region, a characteristic reported for genotype 4 with the possible change of initiation of ORF 3 and ORF 2.
Infection of a goat with a tick-transmitted Ehrlichia from Georgia, U.S.A., that is closely related to Ehrlichia ruminantium

ABSTRACT We detected a novel tick-transmitted Ehrlichia in a goat following exposure to lone star ticks (Amblyomma americanum) from a park in the metropolitan area of Atlanta, GA, U.S.A. Nineteen days after infestation with field-collected adult ticks, the goat developed a fever of two days duration, which coincided with mild clinical pathologic changes and the presence of DNA from a novel Ehrlichia in peripheral blood. The goat transmitted ehrlichiae to uninfected nymphal A. americanum that fed upon the goat, and the ticks maintained the pathogen transstadially. Five months after exposure, immunosuppression of the goat resulted in transient ehrlichemia with transmission of ehrlichiae to feeding ticks. Sequencing and phylogenetic reconstructions of the 16S rRNA, gltA, map1, map2, and ribonuclease III genes suggest the agent might be a divergent strain of Ehrlichia ruminantium, the agent of heartwater, or a new, closely related species. Convalescent serum from the goat reacted with the MAP-1 protein of E. ruminantium and with whole-cell Ehrlichia chaffeensis antigen. DNA from the novel Ehrlichia was detected in 5/302 field-collected adult A. americanum from the park. Our data suggest that A. americanum is a natural vector and reservoir of this Ehrlichia and that domestic goats can be reservoirs The geographic range of the agent and its pathogenicity to humans and livestock needs to be evaluated.
Role of two residues proximal to the active site of Ubc9 in substrate recognition by the Ubc9.SUMO-1 thiolester complex.

The small ubiquitin-like modifier SUMO-1 is covalently attached to lysine residues on target proteins by a specific conjugation pathway involving the E1 enzyme SAE1/SAE2 and the E2 enzyme Ubc9. In an ATP-dependent manner, the C-terminus of SUMO-1 forms consecutive thiolester bonds with cysteine residues in the SAE2 subunit and Ubc9, before the Ubc9.SUMO-1 thiolester complex catalyzes the formation of an isopeptide bond between SUMO-1 and the epsilon-amino group of the target lysine residue on the protein substrate. The SUMO-1 conjugation pathway bears many similarities with that of ubiquitin and other ubiquitin-like protein modifiers (Ubls), and because of its production of a singly conjugated substrate and the lack of absolute requirement in vitro for E3 enzymes, the SUMO-1/Ubc9 system is a good model for the analysis of protein conjugation pathways that share this basic chemistry. Here we describe methods of both steady-state and half-reaction kinetic analysis of Ubc9, and use these techniques to determine the role of two residues, Asp(100) and Lys(101) of Ubc9 which are not found in E2 enzymes from other protein conjugation pathways. These residues are found close to the active site Cys in the tertiary structure of Ubc9, and although they are shown to inhibit the transesterification reaction from SAE1/SAE2, they are important for substrate recognition in the context of the thiolester complex with SUMO-1.
Overreporting of vitamin D deficiency with the Roche Elecsys Vitamin D3 (25-OH) method

Background: Vitamin D deficiency is common. Recently Roche Diagnostics removed their Elecsys Vitamin D3 (25OH) electrochemiluminescence immunoassay (ECLIA) from use, citing deteriorating traceability to the reference method (liquid chromatography tandem mass spectrometry; LCMSMS). We investigated the performance of the Roche assay (2 assay formulations) against an LCMSMS method and the widely used DiaSorin radioimmunoassay (RIA) method. Methods: Two sets of samples from separate populations were assayed for vitamin D. The first set was assayed using three different methods: RIA (DiaSorin) in 2004, polyclonal ECLIA (Roche) in early 2009 and LCMSMS in early 2010. The second set was assayed using polyclonal and monoclonal ECLIA (Roche) and LCMSMS in mid-2010. Results: The correlation of the polyclonal ECLIA with the RIA was poor (ECLIA = 0.45 × RIA + 19, r2 = 0.59, n = 773). LCMSMS results correlated with RIA (RIA = 0.86 × LCMSMS + 4, r2 = 0.69, n = 49) better than with polyclonal ECLIA (polyclonal ECLIA = 0.55 × LCMSMS + 6, r2 = 0.62, n = 55) despite a storage interval of 6 years. In recently collected samples monoclonal and polyclonal immunoassays gave similar results (monoclonal ECLIA = 0.93 polyclonal ECLIA −3, r2 = 0.60, n = 153). The correlation between monoclonal Roche ECLIA and LCMSMS in these samples was very poor (monoclonal ECLIA = 0.31 × LCMSMS + 23, r2 = 0.27). Conclusions: At the time of its removal from the market, the Roche Elecsys Vitamin D3 (25OH) assay showed unacceptable performance, underestimating vitamin D levels. It seems that this bias preceded the introduction of the monoclonal assay. The worldwide distribution of the assay and the duration of this bias likely led to a significant number of patients starting supplementation unnecessarily.
Intracellular calcium ion responses to somatostatin in cells from human somatotroph adenomas

OBJECTIVE Various GH secretory responses to long‐acting somatostatin (SRIH) analogues have been observed during the treatment of acromegalic patients. The effects of SRIH on intracellular Ca2+ homeostasis in human somatotroph adenoma cells has not been examined in detail, and the underlying mechanisms therefore remain to be determined. Using isolated cells from human somatotroph adenomas, we have investigated the SRIH‐induced intracellular Ca2+ responses at a single‐cell level with computerized real time intracellular calcium ion (Ca2+i) imaging.
A putative role for c‐Fos in the pathophysiology of paget's disease

The molecular mechanisms underlying Paget's disease and subsequent osteosarcoma formation are not well understood. In this study, we aim to delineate the function of the c‐Fos oncogene in Paget's disease using transgenic mice, based on previous findings that c‐Fos is highly expressed in Pagetic osteoclasts and that c‐Fos is an essential gene for osteoclast differentiation and skeletal neoplasia. We have generated transgenic mice in which c‐Fos is overexpressed specifically in osteoclasts using the tartrate‐resistant acid phosphatase (TRAP) promoter, and five founder mice have been identified. All transgene‐expressing animals developed severe bone remodeling lesions, some of which progressed to large bone tumors. Histopathologic analysis indicated that the lesions contained a marked increase in the number of osteoclasts that contained a large number of nuclei. Osteoclasts were identified by histochemical staining for TRAP and by in situ hybridization for matrix metalloproteinase‐9 (MMP‐9) expression. Moreover, transgenic osteoclasts, and in some cases, osteoblasts and chondrocytes, expressed high levels of c‐Fos protein as judged by immunocytochemistry. This phenotype of increased osteoclast number and activity, together with an apparently high rate of bone turnover, resembles some characteristics of Paget's disease. These data therefore support an important function for c‐Fos in the Pagetic phenotype, and further support the notion that this gene is important in osteoclastogenesis and in bone remodeling disorders.