Additional studies have constructed and

Additional studies have constructed and analyzed new 27-fold class datasets. For example, with a sequence identity less than 40%, Mohammad et al. (2007) constructed a dataset composed of 2554 proteins belonging to 27-fold classes, proposed structural properties of amino leukotriene receptor antagonist residues and amino acid residue pairs as parameters, and achieved an overall accuracy of 70.5% using 5-fold cross-validation. With sequence identity below 40%, Dong et al. (2009) constructed a 27-fold class dataset (containing 3202 sequences), proposed the ACCFold method, and obtained an overall accuracy of 87.6% using 5-fold cross-validation. Liu and Hu (2010) constructed a new 27-fold class dataset according to the construction of the Ding and Dubchak dataset (2001). This new dataset contains 1895 sequences with a sequence identity below 35%. Motif frequency, low-frequency power spectral density, amino acid composition, predicted secondary structure, and autocorrelation function values were combined as the set of feature parameters. Using the SVM algorithm and the ensemble classification strategy, the overall accuracy in the independent test was 66.67%. Moreover, studies on datasets consisting of 76, 86, and 199 fold classes have demonstrated improvements (Liu et al., 2012; Dong et al., 2009).
In this study, we reorganized the dataset constructed by Liu et al. (2012). According to the biological characteristics, values of the increment of diversity, motif frequency, predicted secondary structure motifs and the average chemical shift information of predicted secondary structure elements were extracted as feature parameters. Based on the ensemble classification strategy, these combined features were used as the input parameter for the Random Forests algorithm. An independent test and 5-fold cross-validation were used to predict the 76 protein fold classes, which resulted in good protein fold identification. The protein folds of the 27-fold class dataset and the corresponding structural classes were also identified, yielding improved results.

Materials and methods

Results and discussion


This work was supported by National Natural Science Foundation of China (30960090, 31260203), The “CHUNHUI” Plan of Ministry of Education, and Talent Development Foundation of Inner Mongolia.

Herba Rhodiolae belongs to the herbaceous perennial plants Rhodiola type and is used in traditional Tibetan medicine. Herba Rhodiolae, which is slightly sweet and bitter in flavor, can adapt to cold, dry or damp environments. As for medicinal applications, it can be used to invigorate blood circulation, stop bleeding, regulate the flow of vital energy and remove obstructions and nourish the blood, support healthy energy levels, make the brain healthy and enhance intelligence. Besides, it can also help to nourish and build the body, relieve fatigue, and treat diseases like senile heart failure, asynodia, diabetes and liver diseases. (Zhang, 1984). The main functions include anti-anoxia, anti-fatigue, anti-aging, anti-toxicity, anti-radiation, anti-cancer and two-way adjustment of the nervus centralis and endocrine system (Bao and Li, 1995). Nowadays, researches on different Herba Rhodiolae mainly focused on the content of chemical compositions extracted in Herba Rhodiolae such as glycosides (Liu et al., 2005). Few research on the physiological and biochemical index measurements of cold resistance is reported, not to mention the comprehensive evaluation of cold resistance for Herba Rhodiolae. However, it is well known that wild Herba Rhodiolae requires rigorous environmental and weather conditions in transplanting, domestication and planting, thus the transplanted Herba Rhodiolae cannot adapt the new environments, leading to an extremely low survival rate (Ashraf et al., 2013a,b). With continuous discovery of the wonderful functions of Herba Rhodiolae, the importance of research on Herba Rhodiolae is emphasized continuously in medicine all over the world recently. At the same time, medicine, health products and foods produced by using Herba Rhodiolae are extensively applied for astronauts, pilots, athletes, divers and people working under special conditions. Herba Rhodiolae medicines and health products are available for sale in Japan, China and many western countries, however, only the wild Herba Rhodiolae cannot meet the market requirements. Therefore, it becomes more and more urgent to study the artificial planting of Herba Rhodiolae. To successfully transplant the Herba Rhodiolae, it is very necessary to deeply understand its cold resistance physiological and biological indexes. Here, in order to pave the way for the wide transplant and production of Herba Rhodiolae, out experiment studies systematically the physiological and biological indexes of four Herba Rhodiolae types under the simulated natural low-temperature conditions at the lab-level.

Little is known about the impact of

Little is known about the impact of injury, violence, and poisoning on maternal or fetal outcomes for women living in low- and middle-income countries (LMICs). Among pregnant women living in LMICs, high risk unintentional injuries such as pedestrian injury may be more common, and the limited use of seat belts and other prevention strategies may increase the risk for vehicle occupants.
There has been limited evaluation of injury-related maternal and fetal deaths during pregnancy in Ghana, where trauma is a leading cause of morbidity and mortality. The objective of this study was to describe maternal and fetal outcomes after trauma at the Komfo Anokye Teaching Hospital (KATH) in Kumasi, Ghana, and identify any association between the prevalence of poor outcomes and maternal risk factors (i.e. demographic and pregnancy characteristics) and injury characteristics.

We conducted a cross-sectional study to investigate adverse maternal and fetal outcomes after trauma in Kumasi, Ghana. Medical records for all females 15years or older who presented to the KATH Emergency Centre (EC) were reviewed to identify women who were recorded as pregnant at the time of presentation after injury.

A total of 29,165 charts were reviewed. Of the total number reviewed, 11,764 (40.3%) patients were women, of which 134 (1.1%) had documented evidence of pregnancy. Maternal demographic, fetal, and injury characteristics of this leukotriene receptor antagonist are presented in Table 1.
Most injured pregnant women were between 18 and 35years of age (81.3%); only 3% were younger than 18. Nearly three out of four injured females were married (71.6%), and most (79.1%) had health insurance. Most women (62.0%) had completed junior secondary education or higher, though 13.4% reported no formal education.
Eleven of the 134 pregnant women evaluated in this study died during hospital admission. There were 37 fetal deaths including still births; almost one in every three fetuses died (27.6%). After including fetal distress (n=13) and premature birth (n=33), a total of 83, or 61.9% of the women included in the study sustained a poor fetal outcome as a result of their injury.
The rate of maternal injury death varied widely by demographic and pregnancy characteristics, including religion, education, insurance status, residence, employment status, and number of live children. Maternal death also varied by injury characteristics, including mechanism of injury and anatomic location of injury (see Supplemental materials for relative proportions and p-values). However in univariate models, the covariates most strongly associated with maternal death were religion, mechanism of injury, and anatomic location of injury.
Poor fetal outcome included fetal distress, premature birth, or fetal death including stillbirth. Poor fetal outcome varied across all examined demographic characteristics, pregnancy and birth characteristics and injury characteristics with the exception of education, insurance status, and alcohol use by partner (see Supplemental materials for relative proportions and p-values). Notably, poor fetal outcomes were more common among younger mothers: 51.8% of mothers with poor fetal outcome after injury were less than 26, compared to only 13.7% of mothers without poor fetal outcome. Furthermore, poor fetal outcome was more common among mothers with non-Akan ethnicity: 74.7% of mothers with poor fetal outcome after injury were of an ethnicity other than Akan, compared to only 21.6% of mothers without poor fetal outcome. Poor fetal outcomes were also more common among non-married women and among women living in non-urban settings. As expected, lower gestational age was associated with a higher prevalence of poor fetal outcome. Crude prevalence ratios are presented in Table 2.
In the multivariable model, maternal age, ethnicity and employment status remained associated with poor fetal outcome (Table 2). Ethnicity other than Akan was associated with a 50% increase in prevalence of poor fetal outcome (aPR 1.5, 95% CI 1.2–1.9). Being unmarried was associated with a 70% increase in the prevalence of poor fetal outcome (aPR 1.7, 95% CI 1.2–2.4). Women Coincidental evolution were unemployed or not in the formal workforce (e.g. unpaid housework) had a lower prevalence of poor fetal outcome, compared with women who were actively employed (aPR 0.6, 95% CI 0.4–0.8).

br Acknowledgements br Introduction In many


In many semi-arid and arid regions of the world, water is limited by quality rather than quantity. Water quality is vital not only because of its impact on the availability of freshwater and human health but also because of its intrinsic value (McCutcheon et al., 1993). The dramatically growing pollution of freshwater resources worldwide during past few decades necessitates comprehensive and proper knowledge of contamination levels and accurate assessment of trends in water quality for protecting this vital resource from pollution as well as for identifying efficient and cost-effective measures to control present and future threats of pollution at local/regional levels (Bartram and Ballance, 1996). In dry regions, most of the water demands are met by groundwater supplies as the factors of mainly leukotriene receptor antagonist growth and climate change cause severe stress to surface water supplies in these areas (Edmunds et al., 2003; Kulkarni et al., 2015). Groundwater quality in an area is greatly controlled by the natural processes (e.g., geology, groundwater movement, recharge water quality, and soil/rock interactions with water), anthropogenic activities (e.g., agricultural production, industrial growth, urbanization with increasing exploitation of water resources) and atmospheric input (Helena et al., 2000; Chan, 2001). Hence, the assessment of groundwater quality at both spatial and temporal scales is imperative for managing this vital resource, especially in water-scarce regions.
Udaipur district (study area of this study), situated in the hard-rock hilly terrain of western India, suffers from severe droughts, low rainfall, high summer temperature and inadequate water resources (Bhuiyan et al., 2006). In western India, droughts occur very frequently and one of the severest historical droughts was seen in the year 2002 (Samra, 2004). At that time, the annual rainfall of the Udaipur district was deviated by as high as −33% from the normal annual rainfall (UNDP, 2002). Consequently, groundwater levels declined significantly in this district (Machiwal and Jha, 2014a), and the diminishing groundwater levels caused deterioration in groundwater quality. Moreover, future climate change and variability may exacerbate the pressure on hydrologic and hydrogeologic systems, which may further deteriorate groundwater quality (Mukherjee et al., 2015). Besides the natural processes, anthropogenic activities prevailing in the area are also degrading groundwater quality at a much faster rate than the former one. The area is well known for its mining and smelting industries, which are probably the potential sources for contaminating surface water as well as groundwater resources. On the other hand, rising population, growing urbanization, and irrigated agriculture are among the other factors responsible for the poor quality of groundwater. The groundwater quality in vast areas is reported to be hard, brackish and with high fluoride levels (CGWB, 2010). This situation calls for an urgent need for developing appropriate strategies to mitigate the effects of groundwater contamination. Management of groundwater resources in hard-rock regions requires understanding of natural and anthropogenic factors, which govern geologic/hydrogeologic processes of the hard-rock aquifer system.
Many conventional tools/techniques for the graphical and statistical interpretation of groundwater quality are described in standard textbooks on groundwater or hydrogeology (e.g., Freeze and Cherry, 1979; Karanth, 1987; Sara and Gibbons, 1991). However, the natural and anthropogenic factors may not be easily distinguished from the chemical composition of groundwater alone. Hence, the need for the application of modern approaches and tools such as multivariate statistical techniques, namely principal component analysis (PCA) and hierarchical cluster analysis (HCA), time series modeling (e.g., trend identification) and GIS techniques has been emphasized for the efficient management of groundwater quality (e.g., Jha et al., 2007; Steube et al., 2009; Machiwal and Jha, 2010). The multivariate statistical techniques offer a valuable tool for the evaluation of spatio-temporal variations and interpretation of complex water quality datasets, apportionment of pollution sources/factors (natural or anthropogenic) and the design of a monitoring network for the efficient management of water resources as well as for finding pragmatic solution to pollution problems (Jeong, 2001; Güler and Thyne, 2004; Valdes et al., 2007). For example, Demirel and Güler (2006) identified anthropogenic factors affecting groundwater chemistry in a Mediterranean coastal aquifer, Mersin–Erdemli basin (Turkey) by applying HCA, PCA and geochemical modeling techniques. The results indicated that anthropogenic factors are responsible for the presence of seasonality in the area where water has been contaminated due to fertilizer and fungicide applications made during early summer season. Cloutier et al. (2008) applied PCA and HCA to identify geochemical processes controlling geochemistry of groundwater in a sedimentary rock aquifer system of the Paleozoic Basses-Laurentides (Canada). Based on loadings of principal components (PCs), the first two PCs were defined as ‘salinity’ and ‘hardness’ of groundwater. Lin et al. (2012) evaluated temporal variability and factors governing shallow groundwater chemistry using analysis of variance, PCA, HCA and geostatistical techniques in the tropical Manukan Island\’s aquifer of Malaysia. Selle et al. (2013) examined spatial and temporal patterns of principal component scores to improve the understanding of processes governing groundwater quality in the Ammer catchment located in southwest Germany. The results indicated influence of land use and geology on the groundwater quality. Kim et al. (2014) employed model-based cluster analysis to differentiate the contributions of natural and anthropogenic factors to the observed groundwater quality in South Korea. Their results demonstrated that bivariate normal mixture model was more robust than multivariate analysis, and provided a better discrimination between anthropogenic and natural groundwater groups.

Some patients undergo radical orchiectomy and postoperative pathology

Some patients undergo radical orchiectomy and postoperative pathology in the clinic to confirm fibrous tumors. leukotriene receptor antagonist Therefore, the correct preoperative diagnosis is important for the treatment of this disease. If scrotal tumors are detected by ultrasonography in patients with normal tumor markers and the tumors cannot be clearly distinguished from the testis, these patients may have a fibrous pseudotumor, and organ-sparing surgery can be performed. Puttemans et al presented color Doppler signal and contrast-enhanced sonography images of a rare paratesticular fibrous pseudotumor and proposed that color Doppler ultrasound and contrast-enhanced ultrasonography could be useful for diagnostics. In addition, magnetic resonance imaging (MRI) played an important role in distinguishing paratesticular fibrous tumors from malignant conditions. The benign lesions have an intermediate-to-low signal intensity on T1-weighted series, whereas they have low signal intensity on T2-weighted series, and it leukotriene receptor antagonist should be noted that the most typical imaging feature of these lesions was the prominently low signal intensity on T2-weighted MRI. Clinical urologists should be well aware of the imaging features of this condition and should properly guide clinicians.
Kressel et al retrospectively analyzed the testicular exams of 670 patients, 12.2% of whom presented with testicular non-germ cell tumors and paratesticular tumors. The majority of patients with fibrous pseudotumors had undergone local excision with reservation testis surgery and had no symptoms of local recurrence, and distant metastases were found during postoperative follow-up. A benign tumor of the tunica vaginalis testis was detected in 5 patients during hydrocele surgery, including 2 cases of fibrous tumors. There was no tumor recurrence after local resection during follow-up for 2 years and 5 years. Most patients with CFT can be cured by local excision and have a good prognosis. To date, recurrence after resection has not yet been reported for CFTs of the tunica vaginalis testis. It is strongly suggested that clinicians implement local excision with testis preservation surgery.


We read with great interest the article by Kapoor et al, “Evolving Strategies in the Treatment of Tuberous Sclerosis Complex-associated Angiomyolipomas (TSC-AML),” providing valuable information on localized and systemic therapy in the management of TSC-AML. However, there was a numeric error in the context. Updated criteria from the 2012 International Tuberous Sclerosis Complex Consensus Conference specify 11 major and 6 minor disease features, instead of 9 minor disease features, used to establish a clinical TSC diagnosis. In addition, current guidelines consider the presence of 2 major features sufficient for a definite TSC diagnosis; however, we should notice that a combination of the 2 major clinical features (lymphangioleiomyomatosis and angiomyolipomas) without other features does not meet criteria for a definite diagnosis.

We thank Cai et al for their comments regarding our article. The authors correctly point out that our study describes the 1998 Tuberous Sclerosis Complex (TSC) Consensus Conference Diagnostic Guidelines. The new updated criteria from the 2012 TSC Consensus Conference removed 3 clinical findings to give a total of 11 major and 6 minor criteria. The following criteria were removed in the 2012 TSC Consensus Meeting: hamartomatous rectal polyps, cerebral white matter radial migration lines, and bone cysts. As discussed in our article, the 2012 TSC Consensus Guidelines require 2 major features or 2 or more minor features for a definite diagnosis; however, lymphangioleiomyomatosis and angiomyolipomas, although major criteria, do not together provide enough risk to provide a definite diagnosis and require another major or minor criteria. Although we believe that clinical diagnostic criteria are important to possibly diagnose TSC, we believe that any individual with clinical suspicion of TSC still requires further assessment including genetic screening and counseling to rule out TSC.

br Discussion PSA screening has led to a

PSA screening has led to a significant increase in detection of clinically localized prostate cancer [3]. As the randomized controlled trials of PSA screening have demonstrated conflicting results [4,5], the U.S. Preventative Services Task Force recommended against PSA screening in all men [6]. We sought to provide population-based evidence to discern further the clinical utility of PSA-based testing.
Our study has 2 principal findings. First, greater frequency of PSA testing in the 5 years before diagnosis of prostate cancer was associated with lower overall and prostate cancer–specific mortality. This is striking, given that our study was limited to men 70 years and older, who are at greater risk for death owing to competing risks. Similarly, surgical treatment of prostate cancer vs. observation has also been shown to improve overall and prostate cancer–specific mortality [19]. Our finding that greater frequency of PSA testing was associated with mortality leukotriene receptor antagonist may be secondary to diagnosing earlier stage, lower-volume disease with fewer metastases. However, although we were unable to adjust for residual confounders such as diet, lifestyle, and body mass index, which may affect overall and prostate cancer–specific mortality [20], a study of baseline PSA drawn from men 50 years or younger demonstrated that though a higher PSA was associated with a greater risk of subsequent prostate cancer diagnosis, no other anthropometric, lifestyle, biochemical, or medical history factors were predictive of a subsequent diagnosis of prostate cancer [21]. Additionally, although the European Randomized Study of Screening for Prostate Cancer demonstrated that PSA screening every 2 to 7 years vs. no screening was associated with a 29% reduction in prostate cancer–specific mortality, it demonstrated no difference in overall mortality [4,22]. Conversely, the Prostate, Lung, Colorectal, and Ovarian trial demonstrated no difference in prostate cancer–specific mortality for annual PSA screening vs. no screening, but it suffered from major limitations including 44% of men obtaining a PSA test before enrollment and 52% in the no screening arm receiving a PSA test during the study [5,23]. Moreover, Crawford et al. [24] demonstrated in subanalyses that men with no or minimal comorbidities who were randomly assigned to screening were less likely to die of prostate cancer than those who were not.
Second, greater frequency of PSA testing was associated with a lower likelihood of metastases at the time of prostate cancer diagnosis. These findings corroborate the expected stage migration observed with widespread PSA screening [25]. The percentage of men with newly diagnosed metastatic prostate cancer has declined from 25% in 1980 to 4% in 2002 [25] with resultant decrease in prostate cancer–specific mortality by 4.1% annually between 1994 and 2006 [26]. It has been previously demonstrated that when compared with younger patients (<75y old), older patients are more likely to present with metastatic disease and prostate cancer–specific mortality despite increased comorbidities [27]. Our results lend support to investigate screening methods that will identify and treat clinically significant prostate cancers before metastases. Although our findings are policy relevant, they must be interpreted in the context of the study design. First, SEER-Medicare is limited to men aged 65 years or older, and our results may not be generalizable to younger men undergoing PSA-based screening. Second, our follow-up was relatively modest, considering prior studies have demonstrated longer follow-up is needed to see a survival benefit [22,28]. In terms of treatment benefit, the Scandinavian Prostate Cancer Study has shown with 15 years of follow-up that prostate cancer–specific mortality and all-cause mortality are lower in men treated surgically compared with watchful waiting [19]. These results were leukotriene receptor antagonist more pronounced in men younger than 65 years. Thus, our results may underestimate the actual survival benefits associated with PSA testing before diagnosis and treatment for prostate cancer. Lastly, although we attempted to control for known predictors for prostate cancer–specific mortality, the findings are hypothesis generating, and it is expected that there will be greater clarification regarding the role of PSA screening with longer follow-up from the current randomized controlled trials. Finally, observational studies reflect practice patterns, and when compared with results from well-conducted randomized controlled trials, they do not appear to overestimate treatment effects nor differ qualitatively [29].

br Materials and methods br Results

Materials and methods

By 2D-DIGE, an almost four-fold up-regulated spot was identified in the sample with known high levels of RegIIIα/PAP transcript compared to the sample with known low levels at about 18–19kDa and pI 5.4 in semi-preparative gels run both at pH ranges 4–10 and 5–7. These properties corresponded with the predicted ones of RegIII proteins (approx. 19kDa and pI 6) as calculated from their primary amino-acid sequences (Fig. 1). The MALDI-TOF/TOF analysis of this spot yielded three peptides belonging to the protein pig RegIIIγ (NCBI Reference Sequence: NP_001138319.1), covering 31% of the mature non-secreted pig RegIIIγ protein sequence (Table 1). This identity was further confirmed when aligning all the primer sequences used in all studies available in the literature (Niewold et al., 2005; Schroyen et al., 2012; Loos et al., 2012; Trevisi et al., 2012). This showed the transcript corresponding to RegIIIγ, rather than RegIIIα. Pig RegIIIγ was 69.5% identical to consensus RegIIIγ, 60.1% to consensus RegIIIα (Fig. 2).
We compared the biochemical properties of the N-terminal (residues 1–25) fragment of the primary amino leukotriene receptor antagonist sequences from consensus RegIIIα and RegIIIγ mature proteins with the pig RegIIIγ (Table 2). Significant differences were observed in pI, aliphatic index and hydropathicity between pig RegIIIγ and the consensus RegIII proteins.
The anti-pig RegIIIγ polyclonal antibody produced against the N-terminal fragment detected on immunoblot a main band of around 16kDa in all analyzed gels, in agreement with the predicted molecular weight of RegIIIγ (Fig. 3). In some samples the main band was accompanied by a less intense band at around 15kDa (Figs. 4 and 5). Samples from the 24h feed and infection experiment with higher mRNA levels of RegIIIγ (Table 3) showed higher levels of RegIIIγ protein than those with lower mRNA levels (P<0.01), but linear regression analysis indicated no relationship between mRNA and protein levels (R2=0.047). We also investigated RegIIIγ band quantity in jejunal mucosa during a natural diarrheal outbreak associated with ETEC and compared with littermates not suffering from diarrhoea. A high individual variability in RegIIIγ abundance was found, and RegIIIγ was relatively more abundant in mucosal scrapings from healthy animals (Fig. 4, pigs 6–10) than diseased animals (Fig. 4, pigs 1–5). This difference was, however, not statistically significant. Concerning the intestinal distribution of RegIIIγ, the four animals from the infection experiment with high expression levels of the transcript were used. RegIIIγ was expressed along the entire small intestine in all four of the animals, the most intense band found at the ileal level and a less intense band in the colon. RegIIIγ was absent from liver except for one of the animals, which showed also high intensity bands in the other tissues studied (Fig. 5).
The aims of the present study was first to identify the most abundant RegIII transcript in the pig intestinal mucosa and then to establish its expression on the protein level during infection. No data were available on the protein level thus far, also because the pig protein did not react with the commercially available anti-human and anti-mouse antibodies (Niewold, 2015). High levels of mRNA do not necessarily mean high concentrations of protein because of the many post-transcriptional and post-translational regulatory events that govern protein production and abundance inside the cell. Therefore, verification of up-regulation of a transcript is needed at the protein level (Greenbaum et al., 2003). The present results show that only one RegIII protein family member was identified among the regulated proteins in the expected gel location (according to RegIII proteins’ theoretical mass and charge) by 2-D DIGE when comparing samples with high and low RegIIIα/PAP transcription levels as determined by qPCR. That protein was confidently identified as RegIIIγ by MALDI-TOF/TOF. These results indicate that RegIIIγ corresponds to the transcript previously known as RegIIIα/PAP, and suggest that it is the most abundant RegIII protein in the pig jejunum. In fact, we did neither detect RegIIIα nor RegIα as a regulated protein in the expected migration area of the 2-DE gel although one report describes the presence of RegIα together with RegIIIγ on the surface of pathogenic bacteria collected from pig intestine (Pieper et al., 2013).

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

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).

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).

Several reports have identified CD also named Siglec or

Several reports have identified CD169, also named Siglec-1 or sialoadhesin, and CD163, two receptors expressed in alveolar macrophages, as the most relevant receptors for cellular attachment, internalization and uncoating of the virus (Duan et al., 1998; Vanderheijden et al., 2003; Delputte et al., 2004; Van Gorp et al., 2008).
CD163 is a member of the family of proteins with scavenger receptor cysteine-rich domains that is expressed on monocytes, some subsets of DC, and specific subsets of tissue macrophages (Sánchez et al., 1999; Flores-Mendoza et al., 2008; Ezquerra et al., 2009; Marquet et al., 2011; Poderoso et al., 2011). CD163 has been shown to colocalize with PRRS virus in early endosomes, having been involved in viral uncoating and genome release in PRRS virus infection (Van Gorp et al., 2008). CD163 interacts with the minor envelope glycoproteins of PRRS virus GP2a and GP4, being the scavenger cysteine-rich domain 5 of CD163 essential for PRRS virus infection (Das et al., 2010). Recently it has been shown that pigs lacking CD163 are protected from PRRS virus challenge (Whitworth et al., 2016).
CD169, a member of the family of sialic leukotriene receptor antagonist binding proteins (Siglecs), shows an expression restricted to some populations of tissue macrophages. Several reports have described a role for CD169 in the attachment and internalization of PRRS virus into cells. In this respect, PRRS virus infection of alveolar macrophages can be blocked in a dose-dependent manner by mAbs to CD169 (Duan et al., 1998; Vanderheijden et al., 2003). Moreover, expression of CD169 in non-permissive PK-15 cells results in binding and internalization of PRRS virus, although uncoating does not take place (Vanderheijden et al., 2003). The interaction between CD169 and PRRS virus occurs through the binding of its N-terminal domain to sialic acids on PRRS virus surface (Delputte and Nauwynck 2004; Delputte et al., 2007b; An et al., 2010). In spite of these results, the importance of CD169 as the primary receptor of PRRS virus has been put in question. MARC-145 cells are infected by the virus although they do not express CD169, and the infection can be blocked by a polyclonal antibody to CD163 (Duan et al., 1998; Calvert et al., 2007). Moreover, expression of CD163 into non-permissive PK-15 and PK032495 cell lines, negative for CD169 mRNA expression, is sufficient to render them fully permissive to PRRS virus infection (Calvert et al., 2007; Wang et al., 2013). Finally, CD169 knockout pigs were infected by PRRS virus showing a clinical outcome, viremia and histopathological signs similar to normal animals (Prather et al., 2013).
In previous studies we have identified in the porcine spleen two main macrophage subsets which differ in the expression of those receptors: a CD163+CD169−/low population located in the red pulp, and a CD163−CD169+ population mostly restricted to the marginal zone and ellipsoids (Poderoso et al., 2011; Alvarez et al., 2014). In this study, we have analysed the permissiveness of splenic CD163+ macrophages to PRRS virus infection. These cells efficiently support the replication of virus, yielding titres slightly higher than alveolar macrophages. Since CD169 is barely expressed in splenic CD163+ macrophages we wondered whether other members of the Siglec family could contribute to the binding of the virus to these cells. Although splenic CD163+ macrophages express Siglec-3 and Siglec-5, and Siglec-3 was found to bind the PRRS virus in a solid phase assay, the binding of virus to these cells and the infection were blocked with antibodies to CD169 but not to Siglec-3 or Siglec-5, suggesting a dominant role of CD169 in virus binding even at very low levels of expression.

Materials and methods


In addition to alveolar macrophages, their primary targets, PRRS virus also infects macrophages in several lymphoid organs. However data on the characteristics of macrophage populations infected by the virus in these organs are scarce. Viral replication in spleen during PRRS virus acute infection has been proved both by virus isolation and viral RNA detection (Xiao et al., 2004; Martínez-Lobo et al., 2011). In the present study we show that splenic CD163+ macrophages efficiently support PRRS virus replication, yielding titres even higher than alveolar macrophages. As these cells have been shown to phagocyte microparticles and process and present antigen to CD4+ T cells in a secondary in vitro response (Poderoso et al., 2011), their infection could contribute to the delayed cellular immune response found in the animals infected by the PRRS virus (López-Fuertes et al., 2000; Xiao et al., 2004). Besides, the replication of PRRS virus in splenic CD163+ macrophages may also facilitate virus spreading to other tissues, as it has been reported for other viruses which infect spleen red pulp macrophages such as dengue virus (Prestwood et al., 2012).

Analyses of population preferences for LTC

Analyses of leukotriene receptor antagonist preferences for LTC options were performed for each of the 20 final vignettes on weighted data, taking into account the sampling design. We present results for the preferred care setting and for the type of care providers, successively, in 6 subsets of vignettes displaying the effect of (1) increasing level of ADL disability: vignettes 2, 4, and 6; (2) incontinence: vignettes 4, 5, and 7; (3) aggressiveness in cognitively intact persons: vignettes 6 and 8; (4) moderate cognitive difficulties: vignettes 2 and 3; (5) severe cognitive difficulties with concerns for security: vignettes 6 and 9; and (6) severe cognitive difficulties with aggressiveness: vignettes 8 and 10. All analyses were conducted using Stata 14.0 (Stata Corp, College Station, TX).
The survey questionnaire was returned by 3133 individuals (88.4% of 3546), of whom 148 (4.7%) stopped before the vignette section and 2985 (95.3%) expressed their opinion on one or more vignettes (final response rate 84.2%). Table 2 shows that nonrespondents were older, lived more frequently without a spouse, had a lower level of education, and reported more frequently difficulties in walking or in BADLs. However, their profile did not differ significantly regarding gender, financial insecurity, the number of reported chronic medical conditions, incontinence, or self-reports of cognitive difficulties.
Characteristics of the Study Population (Weighted Data) and of Respondents Versus Non-Respondents to the Vignette Survey
Population Weighted %
Financial insecurity?
Chronic diseases?
Walking difficulty?
BADL difficulty§
Cognitive difficulty
Urine incontinence?
Reported financial embarrassment or difficulties making ends meet.
Reported treatment or disturbance in the past 12 months for hypertension, coronary artery disease, other cardiac disease, cerebrovascular disease, diabetes, chronic pulmonary disease including asthma, osteoporosis, arthritis, cancer, gastroduodenal ulcer, depression, or Parkinson disease, diagnosed by a physician.
Reported difficulty walking 100 m or inside.
Reported difficulty bathing, dressing, transferring, using the toilet, or eating.
Reported memory troubles affecting daily life or difficulty concentrating or difficulty making decisions, lasting 6 months or more.
Reported urine loss lasting 6 months or more.
Table options
Respondents to the vignette survey completed a total of 55,178 (92.4%) of 59,700 vignettes, giving their choice for an average 18.5 (SD 4.1) of 20 vignettes. The mean was 9.2 (SD 2.2) for the 10 Spouse+ vignettes and 9.2 (SD 2.0) for the 10 Spouse? vignettes. Missing responses did not concentrate on specific vignettes. They ranged from 6.4% to 8.1% for the Spouse+ and from 5.8% to 8.9% for the Spouse? vignettes. In 98.5% of all vignettes, respondents complied with instructions to select one single care option. Most of the recorded double choices concerned Spouse+ vignettes (80.2%, versus 19.8% Spouse?). Respondents hesitated essentially between usual home and sheltered housing options (91% of all double choices) and in relation to vignette 1 (40% of all double choices). In the Spouse+ circumstance, double choices were also more frequent for the vignettes 2 and 3.

In fact Kleinfeld s group has measured serum FFA levels

In fact, Kleinfeld\’s group has measured serum FFA levels in patients undergoing percutaneous transluminal coronary angioplasty decades ago and their results suggest FFA levels may provide a more sensitive measure of ischemia than electrocardiographic measurements [3]. Concerning the role of FFAs in the presence of CAD, several studies have been conducted. Pirro and et al. have conducted a prospective investigation within the Quebec Cardiovascular Study cohort and their results suggest that elevated plasma FFA concentrations are associated with an increased risk of ischemia leukotriene receptor antagonist disease [2]. In recent years elevated plasma FFAs have also been recognized as biomarkers of sudden cardiac death [4] ; [5].
On the other hand, only a few studies have demonstrated the correlation of FFAs with the severity of CAD. Gruzdeva et al. reported that FFA levels increased with increase in the number of diseased vessels and positive correlations were observed between FFA levels and CK-MB in myocardial infarction patients [13]. This study is limited by lack of standard criteria reflecting the severity of CAD. In recent years, a study conducted by Zhao\’s group involved 172 elderly patients who underwent angiography assessed by Gensini score [18]. They concluded that serum FFAs levels were associated with the Gensini score in elderly patients with CAD. However, their study was limited by the small sample size and the role of FFAs in patients of all ages had not been extensively studied. Our study enrolled > 700 patients, a relatively large sample of all ages. We also specifically studied on non-diabetic populations considering the multiple metabolic disorders and cardiovascular risk factors induced by diabetes. In the present study, we found that CAD subjects were tend to have higher FFA levels and plasma FFAs can independently predict the severity of CAD in the Chinese Han population.
In conclusion, we demonstrated that plasma FFAs levels were independently associated with the severity of coronary atherosclerotic lesions in non-diabetic CAD patients.
5. Limitations
There were several limitations in this study. First, the study is from single center. Second, prognostic value of FFAs was not analyzed because this was a cross-sectional study. Finally, the statistical association we found between FFAs and severity of CAD in patients without diabetes was relatively weak and further investigations are needed.
Declaration of interest
The authors have no conflicts of interest to disclose.
This work was partially supported by the National Natural Science Foundation of China (81070171, 81241121), the Specialized Research Fund for the Doctoral Program of Higher Education of China (20111106110013), the Capital Special Foundation of Clinical Application Research (Z121107001012015), the Capital Health Development Fund (2011400302), and the Beijing Natural Science Foundation (7131014) awarded to Dr. Jian-Jun Li, MD, PhD.
1. Introduction
Both diabetes mellitus (DM) and hyperlipidemia play important roles in the development of atherosclerosis and are strong risk factors for cardiovascular events. There is a strong association between glucose and lipid metabolism, and an increase of intestinal cholesterol absorption has been reported in DM patients [1]. Postprandial hyperlipidemia characterized by the accumulation of excess triglyceride (TG)-rich lipoproteins and their hydrolyzed products in a non-fasting state, has been shown to play an important role in the progression or vulnerability of coronary arterial plaque [2]; [3] ; [4]. Lipid metabolism is significantly impaired in DM patients with coronary artery disease (CAD) compared with non-DM patients with CAD [5]. Ezetimibe is a lipid-lowering drug that selectively inhibits intestinal cholesterol absorption by binding to Niemann–Pick C1-like 1 (NPC1L1) protein [6]. The administration of this cholesterol transporter inhibitor has been shown to reduce the serum levels of low-density lipoprotein cholesterol (LDL-C) and fasting TG, especially when used in combination with other drugs, such as statins [7]. Ezetimibe also improves postprandial hyperlipidemia in patients with hyperlipidemia [8]; [9] ; [10].