The large T ags encoded by polyomaviruses including JC

The large T-ags encoded by polyomaviruses, including JC virus, are modular proteins (Fig. 1A). Major domains within T-ag include the N-terminal J-domain, the origin-binding domain (OBD) and the C-terminal helicase domain (reviewed in (Fanning and Zhao, 2009)). Studies of the SV40 T-ag origin-binding domain (OBD) have established that it has multiple roles during the initiation of DNA replication (including site-specific binding to the viral origin (reviewed in (Borowiec et al., 1990; Bullock, 1997; Fanning and Zhao, 2009; Meinke and Bullock, 2012)), helping to melt the central region of the origin (Foster and Simmons, 2010), recruiting cellular proteins to the origin (e.g., (Jiang et al., 2006)) and functioning at the replication fork (Meinke and Bullock, 2012; Meinke et al., 2016)). It is assumed that the OBDs present in other polyomavirus T-ag\’s, such as JCV T-ag, will have very similar functions.
To understand how the T-ag OBDs are able to perform so many functions, we and others have purified the OBDs from several polyomaviruses and determined their structures (Chang et al., 2013; Harrison et al., 2013, 2011; Luo et al., 1996; Meinke et al., 2014, 2016); (reviewed in (Meinke and Bullock, 2012)). These structural studies have established that the individual OBDs contain unique structural features (e.g., (Meinke et al., 2014)). DNA sequencing studies have provided additional evidence that although they are highly homologous, the OBDs contain unique regions. Thus, we reasoned that the purified JCV T-ag OBD could serve as the immunogen for raising a monoclonal antibody that selectively binds JCV, but not SV40, T-ag. Herein we describe the isolation of a monoclonal antibody that selectively detects the OBD from JCV T-ag. We also establish the surface region on the JCV T-ag OBD that is recognized by the anti-JCV OBD monoclonal antibody.

Results
An alignment of the amino histone acetyltransferase residues comprising the JCV and SV40 T-ag OBDs is presented in Fig. 1B. Inspection of this figure establishes that while the OBDs are highly homologous (~81%), there are several unique regions (shown in color). Furthermore, since the structures of both the JCV (Meinke et al., 2014, 2016) and SV40 (reviewed in (Meinke and Bullock, 2012)) T-ag OBDs have been determined, it was histone acetyltransferase apparent which of the unique regions were on the surface of the JCV T-ag OBD (Fig. 1C). Collectively, these observations suggested that the JCV T-ag OBD could be used as the immunogen for raising an antibody against JCV T-ag that does not recognize SV40 T-ag.

Discussion
Monoclonal antibodies against SV40 T-ag have provided a wealth of information regarding polyomavirus DNA replication (e.g., (Gurney et al., 1986; Harlow et al., 1981; Mole et al., 1987)). For instance, the monoclonal antibody Pab416 was shown to inhibit the binding of topoisomerase 1 to SV40 T-ag (Simmons et al., 1996), suggesting architectural features of the initiation complex. In separate studies, a binding site for RPA was mapped to the DNA binding domain (OBD) using additional monoclonal Abs to T-ag (Weisshart et al., 1998).
Monoclonal antibodies that bind selectively to the T-ags encoded by the human polyomaviruses will be equally valuable. Indeed, the need for additional monoclonal Abs is apparent from the increasing correlation between polyomavirus infections and human diseases. For example, between 60% and 80% of adults in the USA and Europe are infected with BK virus (Demeter et al., 2015; Egli et al., 2009; Knowles et al., 2003). Similarly, over 60% of adults are seropositive for MCV infection (Tolstov et al., 2009). Therefore, raising monoclonal Abs that selectively target key viral proteins, such as the T-ags encoded by the human polyomaviruses, remains an important long-term goal. To help meet this need, we developed a protocol for generating monoclonal antibodies that selectively recognize the OBDs from polyomavirus T-ags. Using the purified JCV T-ag OBD as our initial immunogen, we identified and subsequently purified the anti-JCV OBD mAb. Additional experiments conducted with the anti-JCV OBD mAb were used to establish that it recognizes an epitope on the surface of the JCV T-ag OBD that includes residue His 144 (Fig. 7).

Our studies demonstrated that a viral protein is produced

Our studies demonstrated that a viral protein is produced during KHV latency. ORF6 protein may scd1 be the major protein made during KHV latency, since ORF6 RNA is the major transcript detectable during KHV latency by RNAseq analysis (Reed et al., 2015). Many herpesviruses express latency associated proteins during latent infection. During MHV68 latency, the latency associated M2 protein was shown to play a critical role in both the establishment of latency as well as reactivation (Barton et al., 2011; Herskowitz et al., 2008). An M2-null strain of MHV68 replicates with wild-type efficiency in mice but exhibits a dose-dependent defect in the establishment of latency (Jacoby et al., 2002). During EBV latency, up to nine latent proteins can be detected during latency (Kang and Kieff, 2015). EBNA-1 is the major latent protein of EBV and has multiple roles in latency establishment and reactivation (Sivachandran et al., 2012). The latent membrane protein 1 (LMP1) of EBV can block apoptosis and provide growth signals in latently infected cells (Kelly et al., 2006; Le Clorennec et al., 2008; Ndour et al., 2012). EBNA2 is required for protection of the EBV latently infected cells against specific apoptosis stimuli (Lee et al., 2004). KSHV contains a latency locus, which expresses a number of genes and micro RNA during latency, including LANA, vCyclin, vFLIP, kaposin and the twelve viral pre-miRNAs (Chang et al., 2016; Ye et al., 2011). KSHV LANA1 is necessary to maintain nuclear association of latent genomes during replication, as it tethers the viral episome to the chromosome of the daughter cell, thereby ensuring its persistence in replicating cells (Ballestas and Kaye, 2011; Barbera et al., 2004). LANA2 is a latent protein found in scd1 of Multicentric Castleman\’s Disease and primary effusion lymphoma and acts a potent inhibitor of p53 during latency transformation (Rivas et al., 2001). During HCMV latent infection, LUNA (Latency Unique Natural Antigen) has been found to be required for reactivation and immune suppression (Keyes et al., 2012; Mason et al., 2013). It will be interesting to know the function of ORF6 protein and its potential role in KHV latency.
Within KHV infected CCB cells, several species of proteins at different molecular mass were detected by the ORF6 specific antibody, ranging from 68kDa to 130kDa. It is not uncommon for proteins to be detected at an apparently greater molecular mass than predicted. ORF6 protein may have post-translational modifications during late stages of infection. ORF6 protein lacks any predicted coiled coils or a leucine-rich area that would indicate a leucine zipper domain, which is a common dimerization domain in transcription regulator proteins (Landschulz et al., 1988). No difference was observed in samples treated with or without reducing agents, which eliminates the possibility of dimer or heterodimer formation (unpublished data). Post-translation modifications often lead to the increase in the apparent size of a protein. One of the common post-translational modifications is sumoylation with the addition of the small ubiquitin-related modifier, SUMO, and other common modifications being glycosylation or phosphorylation (Anckar and Sistonen, 2007; Garaude et al., 2008; Gravel et al., 2004). Although the theoretical molecular weight of the SUMO proteins is approximately 11kDa, the size increase for each SUMO added on SDS-PAGE is typically in the range of 15–17kDa. In the case of a protein with multiple sumoylation sites, or where SUMO chains form on a lysine target site, correlative increases in molecular weight are expected, sometimes yielding very large shifts in mobility. Multiple bands representing different sumoylation states of the protein are also possible (Park-Sarge and Sarge, 2005, 2009), which may explain the various bands of ORF6 proteins observed at different time post-infections of KHV (Figs. 6, 7).

br Materials and methods br Acknowledgements We want to thank

Materials and methods

Acknowledgements
We want to thank David Pazos for providing with the algorithm to calculate the distribution of viral types among the progeny from the percentages percentages of genomes types determined by sequencing with Illumina Libraries. This study was initially supported by Grant AGL2006-09388/ACU, from the Ministerio de Educación y Ciencia-Proyectos de Investigación Científica y Desarrollo Tecnológico-Programa Recursos y Tecnologías Agroalimentarias.

Introduction
Existing approaches used in the development of vaccines have not worked for HIV-1 (Haynes, 2015) and therefore novel approaches are needed for the development of a successful HIV-1 vaccine. One method is the modeling of immunogens that are based upon target sites of broadly neutralizing antibodies (bnAbs) (Burton et al., 2012; Burton and Mascola, 2015; Haynes, 2015) including the use of the structures of the antibodies themselves to model immunogens (Zhou et al., 2014). The glycans of the V3 and C3 regions form an important part of a target of bnAbs (Sok et al., 2014; Walker et al., 2011) and are therefore of substantial interest as vaccine immunogen models (Pantophlet and Burton, 2006; Walker et al., 2011).
The HIV-1 Envelope (Env) contains a large number of potential N-linked glycosylation sites (PNGs) on its outer surface, with glycans accounting for ~50% of its molecular weight (Korber et al., 2001). These PNGs are usually characterized by an Asn-X-Ser/Thr motif, where X is any amino A-366 except for proline (Gavel and von Heijne, 1990; Marshall, 1974). Glycans are added to the HIV-1 Env by the host cell protein machinery. Therefore, the nature of the glycan found at a particular PNG is dependent on the type of host cell it originates from, the processing that takes place at both the endoplasmic reticulum and Golgi apparatus and on the density of the glycans nearby, which may limit access of processing enzymes (Bonomelli et al., 2011; Hioe et al., 2014). Recent evidence suggests that glycans on Env trimers are consistently under-processed and a high proportion of glycans are Man8–9 GlcNAc2 structures that are important for formation of a compact Env spike (Panico et al., 2016; Pritchard et al., 2015b).
The glycans of the HIV-1 Env protect the virus from neutralization (Moore et al., 2012; Pantophlet and Burton, 2006; Townsley et al., 2016; Wei et al., 2003; Wyatt et al., 1998; Zolla-Pazner et al., 2016), packing together to form a so-called “glycan shield” (Wei et al., 2003). Epitopes protected from neutralization include those on the V3 loop (Binley et al., 2010), a region that is highly susceptible to neutralization by common, mostly narrowly-neutralizing antibodies (Davis et al., 2009; Krachmarov et al., 2001; Vogel et al., 1994), and the CD4 binding site (CD4bs) to which antibody access is heavily constrained by glycans (Binley et al., 2010; Chen et al., 2009; Koch et al., 2003). Removal of particular glycans increases antibody access to the CD4bs (Binley et al., 2010; Koch et al., 2003; Li et al., 2008; McCaffrey et al., 2004; Townsley et al., 2016; Zolla-Pazner et al., 2016) and the V3 loop (Binley et al., 2010; Koch et al., 2003; Li et al., 2008; McCaffrey et al., 2004; Townsley et al., 2016; Zolla-Pazner et al., 2016). Addition/shifting of glycans in the V1, V2, C2, V4 and V5 regions occur in macaques infected with simian/human immunodeficiency virus (SHIV), suggestive of antibody-mediated selection for viruses in which glycans block access to antibody epitopes (Blay et al., 2006).
Initially, the glycans of HIV-1 Env were thought to be almost uniformly unrecognized by antibodies with the exception of the 2G12 epitope (Sanders et al., 2002), leading to terms such as the \”silent face\” of Env (Burton et al., 2005; Wyatt et al., 1998). However, more recently, it has become clear that glycans are very important for the formation of key epitopes such as the V2-glycan site (PG9/16 class of broadly neutralizing antibodies) (Doores and Burton, 2010; Doria-Rose et al., 2015), the epitope targeted by 3BC315 and 3BC176 in gp41 (Lee et al., 2015) and the gp120/gp41 interface epitope (Falkowska et al., 2014; Huang et al., 2014; Scharf et al., 2014). The anti-CD4bs antibody HJ16 is also glycan dependent (Balla-Jhagjhoorsingh et al., 2013). Glycans are also the primary targets for the anti-V3/glycan class of broadly neutralizing antibodies (Julien et al., 2013b; Mouquet et al., 2012; Sok et al., 2014; Walker et al., 2011) that are the main focus of this report.

Therefore the identification of a specific RPE value associated with

Therefore, the identification of a specific RPE value, associated with the VT (RPEVT), may be an effective strategy to guide cardiorespiratory training intensity, particularly from a public health perspective. Thus, the individuals can regulate exercise intensity (e.g., walking or running speed) based on their own perceptual sense. However, previous studies have shown that there is considerable variability in the RPEVT among adults, ranging from 11 to 14, using Borg\’s scale. In general, these studies analyzed the influence of gender, training level, exercise modality, and exercise protocol, evaluating only people with normal body mass.
Particularly in overweight and obese individuals, the identification of the RPEVT has important implications, because exercise performed above the VT in these groups is associated with greater pain sensatio###http://www.GENS-BIO.COM/images/1-s2.0-S1607551X1630153X-gr1.jpg####n, physical discomfort, and feelings of displeasure than in groups with normal body mass. To date, no study has identified the RPEVT in women with different nutritional status. Thus, the aim of this GSK503 study was to identify and compare the RPEVT in normal body mass, overweight, and obese sedentary women.

Methods
The study consisted of three familiarization sessions, followed by a session to evaluate anthropometric measurements, according to the procedures of Gordon et al. and maximal cardiopulmonary fitness. Sessions were separated by at least 48–72 hours. The participants were instructed to abstain from vigorous physical activity, caffeinated products, and alcohol 24 hours before the experimental session. All sessions were conducted with participants wearing athletic apparel and footwear.

Results
The characteristics of the sample are summarized in Table 1. Table 2 shows the results for cardiopulmonary parameters related to the maximal exercise test. The obese group showed lower , at VT, at VT, and %HRmax at the VT than the normal body mass and overweight groups (p < 0.05). There were no differences between the normal body mass and overweight groups (p > 0.05).
Fig. 1 shows the RPEVT during the maximal exercise test in the women with different BMI. There was no difference between groups (p > 0.05): RPE = 12.1 ± 1.4 for normal body mass women; RPE = 12.0 ± 1.6 for overweight women; and RPE = 12.0 ± 1.4 for obese women.

Discussion
The purpose of this study was to identify a specific RPEVT in sedentary women, and to analyze whether there were differences in RPEVT among normal body mass, overweight, and obese groups. We found that the volunteers perceived the exercise intensity associated with the VT as between light and somewhat hard (Borg RPE = 12), independently of BMI. These results are consistent with previous studies that reported a mean RPE value between 11 and 14 at the VT or lactate threshold in different populations (active men and women, sedentary middle-aged men, young trained men, trained cyclists, and type 2 diabetes individuals).
Several studies have demonstrated the benefits of cardiorespiratory training at exercise intensities of close to or at the VT for physical fitness and health. However, in clinical settings, it is often not possible to acquire expensive equipment to monitor cardiorespiratory exercise intensity (e.g., HR monitor, metabolic measurement equipment). Recent studies have shown that perceived exertion is a valid means of evaluating VT, demonstrating acceptable correlation with VT. As such, this method may be recommended to predict maximal functional capacity in sedentary males, young and middle to older-aged individuals who are active or sedentary, and obese women. In addition, previous investigations have shown the efficacy of exercise training involving exercise intensity using perceptually regulated training at a target RPE. Céline et al. found that a 6-week exercise program involving perceptually regulated training at an individualized RPEVT resulted in a mean improvement in VO2max of 10% in healthy young women. Parfitt et al. found that an 8-week exercise program involving perceptually regulated training at an RPE of 13 on the Borg Scale resulted in mean improvements in , mean arterial pressure, cholesterol, and BMI in sedentary men and women. Furthermore, Scherr et al. in a cohort of 2560 participants, reported a very high competency in the assessment of RPE in relation to metabolic (lactate concentrations) and cardiac (HR) intensity parameters, and found that RPE at LT1 was approximately 11 in all groups. From a public health perspective, exercise intensity perceptually regulated at RPEVT may be an effective strategy to be implemented and disseminated for clinical practice. Based on the current results, perceptually regulated exercise intensity at an RPE of 12 may be a practical, simple, noninvasive, and low-cost method to guide exercise intensity at the VT in sedentary women, regardless of BMI classification. Further longitudinal research is necessary to confirm whether cardiorespiratory benefit can be achieved with exercise programming using RPE-based exercise intensity in normal body mass, overweight, and obese women.

Introduction Research on aging is

Introduction
Research on aging is critical because of the enormous healthcare costs associated with supporting a rapidly increasing elderly population. Aging can trigger immune suppression and is associated with increased risk for infectious diseases, autoimmune disorders, and tumor occurrence. Several studies have investigated various agents, such as lifestyle factors (diet, physical activity, and others), which may delay this process.
Researchers have demonstrated that age-specific immunological mechanisms are influenced by gender and that immunosenescence does not affect men and women in the same manner. The age-specific impairment in T Necrostatin 1 is greater in women than in men. Postmenopausal women have decreased T-cell function compared with that in premenopausal women due to the estradiol-to-estrone change in circulating estrogen levels. These sexual hormones affect the activity of the immune system differently.
Regular exercise has many benefits for older people, including improvements in cardiovascular fitness, increases in muscle mass, decreases in the risk factors associated with many life-threatening diseases (e.g., hypertension, cardiovascular diseases, diabetes, and obesity), and strengthening of natural immunity. However, further research on the changes in immune function as a consequence of physical exercise in aging is required. Regular physical activity has beneficial effects on physical and psychological health, as well as on leukocyte function, in young and aged individuals. Many researchers have suggested that regular physical activity decreases the risk of respiratory infection. The “J hypothesis” suggests that low- and moderate-intensity exercise improves immune function, whereas strenuous exercise or overtraining suppresses it. Therefore, regular exercise is of great importance, especially for older people. Nevertheless, no studies have so far evaluated the effect of regular low-intensity training on immune function in middle-aged people.
Leukocyte function and apoptosis are blood markers of immune function. The relationships between age, cytokines, and disease behavior have been studied. Production and release of cytokine play an important role in the first stages of the inflammatory response. Interleukin-2 (IL-2), which is primarily produced by T lymphocytes and natural killer (NK) cells, is a cytokine that participates in many regulatory effects in cells. IL-2 also plays a key role in humoral and inflammatory cellular responses. IL-1ra acts as an antagonist to two other cytokines, namely, IL-1α and IL-1β, which are among the first mediators of the acute-phase response of inflammation.

Materials and methods

Discussion
Aging increases the risk of malignancy and infection. Recently, many researchers have focused on the benefits of regular physical activity on the immune system in young and aged individuals. Although the alterations in the immune system that are triggered by acute exercise have been examined extensively, there are only a few studies about the effect of chronic exercise.
T-cell functions can be affected by aging, and these changes can be evaluated using the in vitro proliferative capacity in response to polyclonal mitogens, such as ConA or phytohemagglutinin (PHA). Shinkai et al observed a reduction in the proliferative response of aged male participants, not only in response to PHA, but also in response to the pokeweed mitogen and alloantigens. A previous study suggested that the age-related reduction in T-cell responsiveness in vitro was accompanied by reduced IL-2 secretion and IL-2R α-chain expression. The aging-influenced cell-mediated immunity may be directly attributed to the age-associated involution of the thymus.
Nieman et al demonstrated that moderate aerobic exercise for 12 weeks increased the NK-cell cytotoxicity or the T-lymphocyte mitogenesis in sedentary women. In addition, a 15-week period of intense exercise in aged rats prevented age-related declines in ConA-induced lymphocyte proliferation, as demonstrated by Nasrullah and Mazzeo. Shinkai et al reported an increase (40–50%) in the proliferative response of active elderly adults compared with that in sedentary ones. In our study, the lymphocyte proliferation in trained MAW was higher than that in untrained MAW. Therefore, the regular low-intensity exercise training improved the immune function. One possible limitation of the present study might be that we did not analyze neutrophil function by migration or phagocytosis capacity.

Binding of purified P TSP to S Typhimurium O antigen

Binding of purified P22 TSP to S. Typhimurium O antigen is weak (Israel et al., 1972), and that of SP6 gp46 is presumably comparable. However, binding of P22 phage to O antigen is irreversible, implying significant cooperativity in binding of multiple TSPs, especially since hydrolysis of O antigen is required for P22 infection (Berget and Poteete, 1980; Iwashita and Kanegasaki, 1973; Schwarz and Berget, 1989). One difference between SP6 and P22 infection is that the P22 TSPs are not known to change orientation during infection. We thus suggest that the initial binding (and/or O antigen hydrolysis) of one SP6 gp46 TSP may only provide energy to disrupt the hand-over-hand garland. However, this step would facilitate binding of other TSPs, ultimately leading to rotation of the entire V-shaped TSP complex. We suggest that a similar process will describe not only SP6 gp47 recognition of the S. Newport O antigen but also K1-5 particles, whose two sets of TSPs differentiate between K1 and K5 capsulated cells (Scholl et al., 2001). There is no experimental information for these two phages.
This process is strikingly different from the mechanism used by T7 or T4 (Hu et al., 2013, 2015a). Binding of a single T7 or T4 long tail fiber to the cell surface has been suggested only to prevent the virion from diffusing away from its target cell, thereby providing time for other fibers to stochastically and non cooperatively dissociate from their SBI-0206965 on the phage tail and/or capsid and interact independently with their receptor.
The major tail proteins of SP6, gp32 and 33, are clearly homologous to those of T7, but an important recent in-depth analysis has revealed that they are homologous to the tails of essentially all podoviruses other than those in the Picovirinae subfamily (Hardies et al., 2016). Because SP6 grows on rough S. Typhimurium strains and the tail appears to make direct contact with the cell surface, whereas the TSPs do not (Fig. 6), tail proteins must be the major determinants of host specificity. The role of TSPs or tail fibers is then to facilitate a productive interaction of the tail with its receptor. By analogy with T7-like phages infecting Yersinia pestis, this receptor is likely the conserved lipid A-KDO region of the LPS (Kiljunen et al., 2011; Zhao et al., 2013). Supporting this conclusion is that a mutant of SP6: SP6coli, which contains four missense mutations in the tail genes 32 and 33, but no alteration in TSP genes, grows well in liquid cultures of laboratory strains of E. coli (Nguyen et al., 2012)(IJM unpublished); these strains have a different truncated LPS outer core than S. Typhimurium but have a comparable inner core.

Material and methods

Acknowledgements
This work was supported by grants R01GM110243 from the NIGMS (to JL and IJM), and AU-1714 from the Welch Foundation (to JL). KH is supported by grant R01GM056141 from the NIH. We are very grateful to Dean Scholl, who both provided S. Newport, S. Heidelberg, and SP6 mutants for this study and also communicated to us prior to publication that S. Newport is a host for SP6.

Introduction
Epstein Barr virus (EBV) is a gammaherpesvirus that is highly proficient in establishing immunoevasive latent infections. Upon infection of B cells, the EBV genome is circularized as a minichromosome and chromatinized by host proteins to establish a restricted latent gene expression program in which only a small percentage of viral genes are expressed. During latency, the virus is subject to low-level spontaneous reactivation (Phan et al., 2016); however, in order to generate a productive lytic infection, the virus must overcome tight regulation to initiate complete reactivation. During latency, the EBV genome is completely chromatinized and subject to suppressive chromatin marks and DNA methylation (Hammerschmidt, 2015; Tempera et al., 2010). Significant efforts have explored changes in the chromatin landscape during reactivation, although it appears these changes are often cell line-dependent and likely consequential, not causal, to reactivation (Flower et al., 2011; Ramasubramanyan et al., 2012; Murata and Tsurumi, 2013). The molecular mechanisms driving the process of reactivation are understood primarily in the context of transcriptional regulation of the BZLF1 promoter (Zp). Numerous factors that bind to Zp have been identified as either positive or negative regulators of BZLF1 transcription, as well as a few cellular factors that contribute to lytic reactivation (McKenzie and El-Guindy, 2015). Understanding the molecular mechanisms dictating the latent to lytic switch may offer therapeutic insight for oncolytic therapy in latency-associated tumors. Further, if these mechanisms are conserved SBI-0206965 among herpesviruses, we could identify therapeutic targets to prevent lytic outbreaks in infected individuals.

BatCoV HKU has been circulating

BatCoV HKU5 has been circulating in bats (Lau et al., 2013). In an epidemiology study over a 7-year period (April 2005 to August 2012), 25% of alimentary specimens of Japanese pipistrelle bats (Pipistrellus abramus) collected from 13 locations in Hong Kong were positive for this virus (Lau et al., 2013), indicating that it might target gastro-intestinal tissues. However, BatCoV HKU5 virus has not been isolated and cultured successfully, which is an obstacle to virus transmission research. The problem is largely due to a lack of suitable cell lines for BatCoV HKU5 virus. The emerging but puzzling question is whether this virus could infect humans or not.
An infectious clone of BatCoV HKU5 containing the ectodomain from the SARS-CoV S protein was constructed through reverse genetics and synthetic-genome design, and the recombinant virus replicates efficiently in cell culture and in young and aged mice (Agnihothram et al., 2014). In addition, the key proteins for virus replication, such as the 3C-like protease, polymerase, and exonuclease of BatCoV HKU5 display high amino leukotriene receptor antagonist sequence similarity to those in MERS-CoV, indicating that once the genome of BatCoV HKU5 is released into host cells, genome replication, virus particle assembly, and release can readily occur. Therefore, the receptor would be the last barrier for BatCoV HKU5 to infect humans. Our data show that BatCoV HKU5-CTD does not use hCD26 as a receptor, though it folds into a very similar structure as MERS-RBD/CTD and HKU4-RBD/CTD. leukotriene receptor antagonist In other words, the cellular receptor of BatCoV HKU5 is still a mystery that requires further study.
Evolutionally, BatCoV HKU5 S protein is more diverse than BatCoV HKU4 S protein (Lau et al., 2013), and various deletions in loop 1 (Fig. 3D and Fig. 6) have been sequenced. This indicates that BatCoV HKU5 is able to generate variants to occupy new ecological niches and might acquire the ability to bind to hCD26 by accumulating mutations and ultimately cause human respiratory infections like MERS-CoV and SARS-CoV. Accordingly, it is very important to perform long-lasting surveillance of BatCoV HKU5 evolution, especially the variety of S protein in the event that the virus breaks the inter-species and/or inter-tissue transmission barriers.

Materials and methods

Acknowledgements
This work was supported by the National Natural Science Foundation of China (NSFC, Grants 81461168030 and 31502078), the Key Research and Development Program of Ministry of Science and Technology of China (MOST, Grant 2016YFC1200300) and the China National Grand S&T Special Project (Grant 2014ZX10004-001-006). G. L. is supported in part by NSFC (Grant 81522026 and 31400154) and the Sichuan Outstanding Youth Science & Technology Funding (Grant No: 2016JQ0001). Y.S. is supported by the Excellent Young Scientist Program of the Chinese Academy of Sciences (CAS) and the Youth Innovation Promotion Association CAS (2015078). H.S. is supported by the Youth Innovation Promotion Association CAS (2017117). K.Y. Y. is partly supported by the NSFC/RGC Joint Research Scheme (N_HKU728/14) and the Theme-Based Research Scheme (T11/707/15) of the Research Grants Council, Hong Kong Special Administrative Region. G.F.G. is a leading principle investigator of the NSFC Innovative Research Group (Grant 81621091).

Introduction
Tick-borne encephalitis (TBE) is a severe infectious disease affecting the central nervous system (CNS) of humans. TBE is caused by TBE virus (TBEV), a member of family Flaviviridae, genus Flavivirus (Lindquist, 2014; Mansfield et al., 2009). Over the past few decades, TBE has become a growing public health concern in Eurasia, with more than 10,000 clinical cases of TBE, including numerous deaths, reported every year (Bogovic and Strle, 2015). Clinical presentation of TBE ranges from fever or meningitis to more severe meningoencephalitis or encephalomyelitis. Long-term neurological sequelae are common after TBE and usually involve paresis, ataxia, or gait disturbance (Bogovic et al., 2010; Růžek et al., 2010).

Some cytokines are produced by monocytes

Some cytokines are produced by monocytes and/or macrophages in the subendothelial adipose tissue and about 30% of IL-6 is released by visceral adipose tissue. In addition to adipose tissue, skeletal muscle is also an important source of cytokines. TNF-α is involved in insulin resistance as it is overproduced by adipose tissue in obese patients. CRP, an acute phase protein, is produced by the liver in response to inflammatory cytokines such as IL-6 and TNF-α and can be modulated by exercise, depending on the intensity and duration of the sessions. According to our results, higher levels of inflammatory markers such as TNF-α and hs-CRP (p < 0.05) were observed in overweight and obese patients after training. Martín-Cordero et al, in a study evaluating obese rats, showed that regular exercise was able to improve the release of TNF-α by peritoneal macrophages, improving the balance between inflammatory markers, reaching the behavior of lean healthy rats. Another interesting study that evaluated overweight men after 16 weeks of training did not observe differences in cytokine levels. However, the authors indicated a change in hs-CRP levels, suggesting an increase in this variable during the post-training period. It is well known that CRP has an acute response to exercise and it can remain increased for several days following the last session of exercise. In our study, blood was collected 48 hours after the last exercise session, which may reflect the increased level of hs-CRP after training. Furthermore, there is evidence that training programs performed weekly at high frequencies can be very stressful, increasing some inflammatory markers. Therefore we suggest that the short duration (3 months) of the intervention used in our study may have induced an acute low-grade 4-ethylphenyl sulfate in patients, as the progression of exercises and the number of series/repetitions increased rapidly, increasing the levels of inflammatory markers such as CRP and TNF-α.
Thus a long period (more than 3 months) of moderate exercise could regulate inflammation. However, our data contradict previously published studies which showed a reduction in pr###http://www.apexbt.com//media/diy/images/struct/B1406.png####o-inflammatory biomarkers after the performance of different types of exercise in obese and sedentary patients, highlighting the anti-inflammatory role exerted by exercise. Balducci et al, in a study of 12 months of exercise training in patients with type 2 diabetes mellitus and metabolic syndrome, found decreased levels of hs-CRP, TNF-α, IL-6, and IFN-γ. Jorge et al submitted obese type 2 diabetic patients to aerobic, resistance, and concurrent training for 12 weeks and observed reductions in the hs-CRP levels in all groups. Moreover, studies involving the effect of different modalities of exercise on inflammation are still controversial, requiring further research in tissue and molecular levels to clarify the correct effects of physical activities on immunity and the precise cell source of cytokines.
This study investigated the frequencies of CD3+CD4+ and CD3+CD8+ T lymphocytes as well as the percentage of HLA-DR+ lymphocytes and monocytes in obese and overweight patients after 12 weeks of concurrent training. There is no published data correlating the expression of CD4, CD8, and HLA-DR in peripheral blood cells with exercise in obese/sedentary patients. Fu et al, in an animal model, have shown that moderate exercise training prevents a decrease in CD4+ T cells. Our study was characterized by exercise of moderate intensity (50–75% Vo) with linear progression, which could help to prevent the decrease in CD4+ T lymphocytes. Therefore moderate exercise appears to improve the defense mechanisms of the body by increasing the function of leukocytes and decreasing susceptibility to disease, whereas intense exercise seems to weaken them. Shimizu et al, evaluating 12 weeks of concurrent training in elderly patients, observed increased frequencies of CD3+CD28+ T lymphocytes after the training period, indicating a high proliferation of cytotoxic and senescent T cells.

br Science and the state

Science and the state
In their examination of scientific debates in early modern England, Shapin and Schaffer famously asserted, ‘solutions to the problem of knowledge are solutions to the problem of social order’ (1985, p. 332). With these words, they were arguing for the broader significance of science studies. Their seminal book illustrated how approaches to science reflected broader socio-political trends and how science also supported or subverted approaches to social and political governance more broadly.
Shapin and Schaffer specify that science occupies the same ‘terrain’ as politics in three key ways. Firstly, scientific practitioners and their activities contribute to creating and maintaining the polity in which they operate. For example, the fact that the Soviet leadership was saturated with engineers and technical specialists may have propagated and sustained the state\’s traditions of social engineering and rigid long-term planning. Secondly, the products of science become parts of the political activity of the state. An obvious example here, explored in detail by historians of Soviet science, is the role of nuclear weapons in maintaining a balance of power between the USA and the Soviet Union during the Cold War (Roberg, 1998). Thirdly, scientific endeavors are supported (financially and politically) and valued ‘insofar as the state or its various agencies see a point in them’ (Shapin & Schaffer, 1985, p. 339, see also p. 332). A greater push for commercial relevance of science (nanotechnology, innovation) is one clear, recent manifestation of this in Russia today (Connolly, 2013; Graham & Dezhina, 2008). These clear manifestations of the ‘shared terrains’ identified by Shapin and Schaffer suggest that seeking to understand broader political trends in Russia through the lens of science and knowledge projects is a fruitful and legitimate avenue for analysis, and the article takes its point of departure in this assertion.

Skolkovo: discourses on the role of the state, the future and the foreign
In many ways, the Skolkovo project was the epitome of Medvedev\’s modernization ‘campaign’. The idea of modernization has deep historical ceramide manufacturer in Russia – from Peter the Great\’s dream of a Europeanized Russia to the Soviet attempt at a leap from a peasant society to an urban industrial one. Under Medvedev\’s presidency (2008–2012), the notion of modernization gained new currency in Russian politics. In autumn 2009, Medvedev published a liberal manifesto, Forward Russia! (Medvedev 2009). Its centerpiece was the idea that Russia had been increasingly lagging behind developed countries in science, technology and economics due to corruption and dependency on natural resources.
What modernization was actually supposed to mean as a vision for Russia\’s future and as an engine for economic, social and political change remained ambiguous. In particular, the question of whether modernization was also to include a liberalization of Russia\’s politics was at one point hotly contested, although optimism on that point has largely faded (Devyatkov & Makarychev, 2012; Malinova, 2012; Marganiya, 2010; Zaostrovtsev, 2010). As the case study in suprachiasmic nucleus (SCN) section shows, the Skolkovo project itself embodies the ambiguities and aspirations of the modernization project more broadly.
The Skolkovo project was launched in Medvedev\’s 2009 address to the Federal Assembly. Conceptual work and attending legislation then proceeded at what was proudly described by Kremlin top aide Arkady Dvorkovich as a ‘record fast’ pace (Emel’yanenkov, 2010a), with a number of concessions meant to address constraints to innovation in Russia. A laundry list of obstacles to innovation-based economic development includes: unfavorable legal and bureaucratic environment around imports/exports, unfavorable taxation regime, low investment in research and development (R&D), weakly developed property and intellectual rights, low domestic demand for the products of innovation, lack of coordination between government bodies, poor infrastructure and low levels of access to finance (Connolly, 2011; Dezhina, 2011; Klochikhin, 2012; Spiesberger, 2011). The Rossiskaya Gazeta (RG) coverage analyzed below included voices, especially from scientific establishments and private business associations, concerned about the feasibility of the project because of these existing constraints (Fyodorov, 2010; Kalysheva, 2011a, 2011b; Petrov, 2010).

Russian companies are also looking to

Russian companies are also looking to invest in some other areas of the North Korean economy. The principle challenge facing them is that they thapsigargin need to be confident that these projects will be profitable, and that they will then be able to get these profits out of the DPRK. The case of Egyptian company Orascom\’s investment within the telecom sector of North Korea has often been cited as the most successful example of foreign investment in the DPRK. However, because of international sanctions, and after years of profitable operation, Orascom are now experiencing severe difficulty in exchanging its income from local currency into dollars, and with getting these funds out of North Korea. This high profile example could seriously discourage potential private Russian investors who have not yet gained enough confidence to invest in the DPRK.

Interregional economic ties: more potential exists
The most dynamic area of interregional cooperation is attracting North Korean labour for temporary work in the territory of the Russian Federation. Implementation of federal and regional programmes in the development of the Russian Far East has led to a significant increase in the number of workers from the DPRK. In 2010 about 21,000 North Korean citizens worked in Russia in such spheres as construction, agriculture, forestry, health care, fishing and light industry. In 2013, Russia increased the permits quota for foreign workers from North Korea to 35,000. This area of cooperation is highly beneficial both for Russia (disciplined, law-abiding and inexpensive workers help alleviate labour shortages in Siberia and the Far East) and North Korea, which receives significant currency earnings. As a result of increased cooperation between North Korea and the Russian Federation passenger traffic of the North Korean aviation company Air Koryo between Vladivostok and Pyongyang grew by 22% in the first half of 2014. In 2015, the amount of North Korean workers in Russia increased to 47,364 people, which is 27% more than in the previous year. As a consequence, the DPRK became the 3rd most significant foreign country, after China and Turkey, in terms of the number of work permits issued in Russia.
North Koreans have recently shown an increased interest in the implementation of agricultural projects in the Russian Far East. Since 2011, various options for cooperation with Amur region including joint projects for setting up dairy and beef farms, as well as cultivation of grain and soybeans have been discussed. In mid-2013, the Consul General of the DPRK in Nakhodka at a meeting with the Governor of Primorsky Territory said that North Korea plans to invest $1 million in processing corn and soybeans as well as to consider joint projects in cattle breeding in Primorye. North Koreans currently have experimental agricultural enterprise in the Dalnerechensk district of Primorsky Territory.
In 2014, the DPRK officials announced they were interested in renting 10,000 hectares of land in Khabarovsk region to grow vegetables, breed cattle and set up processing enterprises using Korean labour and equipment. There were reports of possible involvement of investors from the Middle East in the financing of these projects. Most of the products produced in Russia at North Korean agricultural enterprises would then be exported to the DPRK to improve the food situation.

The future of economic cooperation between Russia and North Korea: problems and prospects
The project of supplying Russian gas to South Korea through North Korea has been discussed for more than 20 years. After the construction of the “Sakhalin-Khabarovsk-Vladivostok” main gas pipeline was completed in 2011, the infrastructure for building an extension to the Korean peninsula was ready. North Korea for its part, agreed to provide land for the construction of a gas pipeline, and Russia and South Korea came close to discussing commercial gas supply contracts. However, in 2012, the parties were unable to agree on the price of gas. After that, the communication on this subject was frozen but neither side said that cycle was buried forever.