Development of Monoclonal Antibodies Against Human IRF-5 and Their Use in Identifying the Binding of IRF-5 to Nuclear Import Proteins Karyopherin-α1 and -β1

PURPOSE: IRF-5 is a direct transducer of virus-mediated and TLR-mediated signaling pathways for the expression of cytokines and chemokines which form homodimers or heterodimers with IRF-7. However, direct IRF-5-specific monoclonal antibodies (mAbs) are not available at present. These could be used to further evaluate the functions of IRF-5. In this study, we produced and characterized three mouse mAbs to human IRF-5. The binding of IRF-5 to nuclear import proteins was first identified using a mAb. MATERIALS AND METHODS: His-tagged human IRF-5 protein spanning amino acid residues 193-257 was used as an antigen and three mAbs were produced. The mAbs were tested with ELISA, Western blot analysis (WB), immunofluorescent staining (IF), and immunoprecipitation (IP). In addition, the nuclear import protein which carried phosphorylated IRF-5 was identified using one of these mAbs. RESULTS: MAbs 5IRF8, 5IRF10 and 5IRF24 which reacted with the recombinant His-IRF-5(193-257) protein were produced. All mAbs bound to human IRF-5, but not to IRF-3 or IRF-7. They could be used for WB, IF, and IP studies. The binding of phosphorylated IRF-5 to karyopherin-alpha1 and -beta1 was also identified. CONCLUSION: Human IRF-5-specific mAbs are produced for studying the immunologic roles related to IRF-5. Phosphorylated IRF-5 is transported to the nucleus by binding to nuclear import proteins karyopherin-alpha1 and -beta1.ope

538 A.D. and the Transition from Pagan Roman Empire to Holy Roman Empire: Justinian’s Metamorphosis from Chief of Staffs to Theologian

The year 538 A.D. became the turning point in the history of the Roman Empire since so many aspects on political, administrative and economical levels were already switched off that when Justinian declared himself to be a theologian from this year and no longer a soldier, he crossed the barrier of his mandate between what is purely civil obligation and what is religious obligation, similarly to Constantine before, and entered in competition with the papal function and this role is evidence of Justinian’s ongoing caesaro-papism. The quest for unification of the empire by unification of the church, the fever for church-building projects with his wife Theodora, the persecution of enemies of the church and heretics, his disdain with the Sabbath although his second name was Sabbatini, his support for suppressing any eschatological fever in line with the church fathers and Oecumenius and yet trying to build the ‘Kingdom of God’ on earth, all this indicate the problem 538 was for the Roman Empire and the Catholic Church. Archaeological and historical original sources of Justinian and contemporaries of popes, biographer of Justinian and a commentator on Revelation (Oecumenius) are very revealing of these times and the shift or transition of what belonged to the Roman Empire handed over since 538 A.D. to the church and the papal function. The Code of Justinian was a persecuting instrument. Justinian upheld the supremacy of the papacy. He permitted through the Council of Orleans actions to be done on Sunday that Constantine prohibited like travel and preparation of food and cleaning the house. In Novellae CXLIV Justinian instituted a Seventh-day Sabbath persecution. He changed the times and laws ad hoc as his Novellae XLVI and coins of 538 A.D. (XII year) indicate. Private gatherings were persecuted. He had church-manual laws. Justinian studied Systematic Theology on the nature of Christ and wrote homiletical rules for preachers. He gave textcritical advice to Jews and condemned their doctrinal deviations. This theological hobby of the ruler of the once mighty Roman Empire was to be taken over by a more theological competent power that would eventually lead to papal-caesarism until the unsettling of this new aggrandizing paradigm in 1798 by Napoleon. The prophetic embedding of the 1260 days as “years” prophecies in both Daniel 7 and Revelation 12 definitely started in 538 A.D. contrary to W. Spicer’s (1918) suggestion of 533 or 538 as two alternative dates or any other dates suggested by other scholars in the history of interpretation in historicism. It is also not just a case of history of interpretation hermeneutics but data solidly supported by archaeology, iconography and original historical sources that coincides with the parameters provided by exegesis of the rest of the Books of Daniel and Revelation added with the exegesis of the detail of the passages under consideration. A necessary ingredient for the historical researcher remains to be the faith that God can predict the future and He did and that the data as well as the prophecies of the Biblical Text are evidence of that

Clinical Features of Probable Cluster Headache: A Prospective, Cross-Sectional Multicenter Study

Background: Epidemiological data of probable cluster headaches (CH) are scarce in the relevant literature. Here, we sought to assess the prevalence and clinical characteristics of probable CH in comparison with definite CH.Methods: Data used in this study were obtained from the Korean Cluster Headache Registry (KCHR), a prospective, cross-sectional, multicenter headache registry that collected data from consecutive patients diagnosed with CH.Results: In total, 159 patients were enrolled in this study; 20 (12.6%) were diagnosed with probable CH. The most common unfulfilled criterion in patients with probable CH was the duration of attack, which was found in 40% of patients with probable CH. Among clinical characteristics, the number of autonomic symptoms tended to be lower in probable CH than in definite CH (1.7 ± 1.2 vs. 2.4 ± 1.5, p = 0.051) and conjunctival injection and lacrimation showed an increased odds ratio (OR) [OR = 3.03; 95% confidence interval (CI): 1.03–8.33] in definite CH. The groups did not differ with regard to baseline demographic characteristics, disability, impact on life, or treatment response.Conclusions: Probable CH is relatively common among CH disorders, with a clinical impact similar to that of definite CH

Specific Detection of Influenza A and B Viruses by CRISPR-Cas12a-Based Assay

A rapid and accurate on-site diagnostic test for pathogens including influenza viruses is critical for preventing the spread of infectious diseases. Two types of influenza virus, A and B cause seasonal flu epidemics, whereas type A can cause influenza pandemics. To specifically detect influenza A (IAV) and B (IBV) viruses, we developed a clustered, regularly interspaced, short palindromic repeats (CRISPR) and CRISPR-associated (Cas) system-based assay. By coupling reverse transcription recombinase polymerase amplification (RT-RPA) and reverse transcription loop-mediated isothermal amplification (RT-LAMP), a CRISPR-Cas12a DNA endonuclease-targeted CRISPR trans-reporter (DETECTR) detected IAV and IBV titers as low as 1 × 100 plaque forming units (PFUs) per reaction without exhibiting cross-reactivity. Only 75 to 85 min were required to detect IAV and IBV, depending on isothermal nucleic acid amplification methods, and results were verified using a lateral flow strip assay that does not require additional analytic equipment. Taken together, our findings establish RT-RPA and RT-LAMP-coupled DETECTR-based diagnostic tests for rapid, specific and high-sensitivity detection of IAV and IBV using fluorescence and lateral flow assays. The diagnostic test developed in this study can be used to distinguish IAV and IBV infections, a capability that is necessary for monitoring and preventing the spread of influenza epidemics and pandemics

Impact of cluster headache on employment status and job burden: a prospective cross-sectional multicenter study

Abstract Background Cluster headaches (CH) are recurrent severe headaches, which impose a major burden on the life of patients. We investigated the impact of CH on employment status and job burden. Methods The study was a sub-study of the Korean Cluster Headache Registry. Patients with CH were enrolled from September 2016 to February 2018 from 15 headache clinics in Korea. We also enrolled a headache control group with age-sex matched patients with migraine or tension-type headache. Moreover, a control group including individuals without headache complaints was recruited. All participants responded to a questionnaire that included questions on employment status, type of occupation, working time, sick leave, reductions in productivity, and satisfaction with current occupation. The questionnaire was administered to participants who were currently employed or had previous occupational experience. Results We recruited 143 patients with CH, 38 patients with other types of headache (migraine or tension-type headache), and 52 headache-free controls. The proportion of employees was lower in the CH group compared with the headache and headache-free control groups (CH: 67.6% vs. headache controls: 84.2% vs. headache-free controls: 96.2%; p = 0.001). The CH group more frequently experienced difficulties at work and required sick leave than the other groups (CH: 84.8% vs. headache controls: 63.9% vs. headache-free controls: 36.5%; p <  0.001; CH: 39.4% vs. headache controls: 13.9% vs. headache-free controls: 3.4%; p <  0.001). Among the patients with CH, sick leave was associated with younger age at CH onset (25.8 years vs. 30.6 years, p = 0.014), severity of pain rated on a visual analogue scale (9.3 vs. 8.8, p = 0.008), and diurnal periodicity during the daytime (p = 0.003). There were no significant differences with respect to the sick leave based on sex, age, CH subtypes, and CH recurrence. Conclusions CH might be associated with employment status. Most patients with CH experienced substantial burdens at work