Monthly Archives: August 2018

Cause specific mortality and morbidity outcomes in the

Cause-specific mortality and morbidity outcomes in the systematic review and meta-analysis are reported using the International Classification of Disease (ICD 10) nomenclature and hierarchy (Supplementary File S5). A total of 13 publications were suitable for only the systematic review, including study IDs 56–61, which present risk estimates comparing two temperatures (Supplementary Table 2). The risk of cardiovascular and cerebrovascular outcomes, including cerebral infarction, cerebral haemorrhage and ischemic heart disease increased with heat [study IDs 56–61] and cold exposure in most studies [study IDs 56–61]. Only Pan, et al. report a 0.73 lower odds of cerebral haemorrhage at 32 versus 28°C. Respiratory deaths decreased by 2.5% with heat exposure in Chicago, USA at 29 versus 18°C purchase Dig-11-ddUTP [study ID 59]. Cold exposure increased the risk of allergy [study ID 61] and respiratory mortality and morbidity [study IDs 58,59,61]. A non-linear relative risk of mental health emergency department visits ranging from 1.05 to 1.09 at 25°C was observed across three regions of Quebec, Canada [study ID 36]. Studies applying DTR [study IDs 4,11,23,27,29,34], an exposure metric not comparable to temperature or apparent temperature was included only in the systematic review. Cardiovascular mortality and morbidity relative risk increased in all but the 75+; 0.95 (95% CI 0.06–1.84) [study ID 4], and 65–74 group; 0.99 (0.99–1.0) [study ID 27] per 1°C increase in DTR. Heart failure morbidity increased dramatically in Hong Kong, China [study ID 29]. Interestingly, the risk of respiratory morbidity (all respiratory, respiratory tract infection) increased with a 1°C increase in DTR [study IDs 23,27,34], whereas mortality decreased (Basu et al., 2005). In Taiwan [study ID 27], the 75+ group exhibited slightly elevated risks per 1°C increase in DTR for renal; 1.02 (1.0–1.04) and digestive; 1.04 (1.02–1.05) morbidity compared with the 65+ age group.
We report mortality meta-estimates (where k>2) for ischemic heart disease (ICD-10 codes I20–25), all cardiovascular (I00–99), all cerebrovascular (I60–69), and all respiratory outcomes (J00–99). Morbidity meta-estimates (k>2) are presented for ischemic stroke (I63), intracerebral haemorrhage (I61), myocardial infarction (I21–23), purchase Dig-11-ddUTP pectoris (I20), heart failure (I50), asthma (J45–46), pneumonia (J09–18), diabetes mellitus (E10–14), acute renal failure (N17), intestinal infectious (A00–99), heat-related outcomes (E70–90, T66–78 and R00–99), all cardiovascular (I00–99), all cerebrovascular (I60–69), and all respiratory outcomes (J00–99). In addition, we present an ‘overall’ estimate for both mortality and morbidity outcomes, an amalgamation of the subgroups presented by individual studies including the ‘all’ category. The frequency count of each disease subgroup in the meta-analysis is given in Fig. 3. All cardiovascular disease (CVD) and all respiratory disease (RD) outweighed other mortality subgroups. A greater range of disease groups were represented by morbidity outcomes, including all RD and all CVD, followed by cerebral infarction, myocardial infarction, heart failure, all genitourinary disease, intestinal infection and diabetes mellitus. Threshold temperatures were selected by individual studies based on study location. Lag times, presented as cumulative or single day lags describing the delay between exposure and outcome, varied with high and low temperature. Heat lags were shorter than cold lags, generally between lag 0–1 to 0–3days prior to the event. Cold lags ranged between to 30days.
We observed a striking increase in the risk of all mortality outcomes, including cerebrovascular, cardiovascular and respiratory outcomes (Table 2 and forest plots in Supplementary File 6). The greatest risks were for heat-induced CVD and RD, and cold-induced RD mortality. Cerebrovascular (CBD) risks also increased with cold exposure. No protective effect (risk reduction) was found for any mortality outcome; only ischemic heart disease risk was statistically insignificant; 0.45% (95% CI −0.01–0.91) per 1°C decrease in temperature. Compared to the universally elevated mortality risks, temperature-related morbidity risks were mixed for CBD and CVD outcomes (Table 3 and forest plots in Supplementary File 6). In warm periods, the risk of intracerebral haemorrhage, myocardial infarction and all CBD morbidity reduced per 1°C increase in temperature. In winter, the risk of morbidity from angina, heart failure, all CVD, and all CBD reduced per 1°C decrease in temperature. Heat exposure led to an increase in RD morbidity by 2.76% (1.51–4.03). A statistically significant increase was noted for heat-induced diabetes mellitus 1.02% (0.43–1.62) and an even greater risk for heat-related overall genitourinary morbidity 2.12% (1.65–2.59). A 1°C increase in temperature resulted in elevated risks for overall infectious and heat-related morbidity. The greatest statistically significant risk was associated with pneumonia; 1°C reduction in temperature caused a 6.89% (1.20–12.99) increase in morbidity in cold periods.

A secondary outcome of interest was the

A secondary outcome of interest was the arrest to birth interval. Out of 34 babies for whom the information is known, 4 newborns (11.8%) were delivered within 5min or less, 12 (35.3%) delivered from 5 to 11min, 8 (23.5%) delivered from 11 to 21min, and 10 (29.4%) delivered more than 21min after the maternal cardiac arrest.

Discussion
While one would expect most pregnant women to experience Dig-11-ddUTP damage within a few minutes of cardiac arrest, that was not observed in this case series. The majority of women survived without injury even after resuscitation for more than 5min. Our findings are consistent with case series review of Einav et al. who found an overall maternal survival rate of 54% and a neonatal survival rate of 64% despite the fact that the average time from arrest to birth was approximately 16min and only 7% of the population had a Cesarean birth initiated within 4min (Einav et al., 2012). Contrary to our study, the published reports examined by Einav et al. included women who suffered a cardiac arrest and a perimortem Cesarean birth and those who did not have a Cesarean birth. Their analysis showed Dig-11-ddUTP a higher survival in women who did not have a perimortem cesarean birth compared to those who did. However, as acknowledged by the authors, these findings may very well be biased by confounders (e.g., less severe patients have a return to spontaneous circulation before cesarean delivery could be performed; longer arrests are associated with worse outcomes). Hence, such data should not be interpreted to mean that women who suffer a cardiac arrest should not be delivered. Though similar data was used both for Einav\’s assessment and that of the authors of this study, our dissimilar conclusions on the timing of delivery is based on a different way to analyze these data. Einav\’s recommendation to deliver within 10min of a documented cardiac arrest is based on the fact that only the ten-minute interval was examined in the regression model used, whereas our study performed an assessment of a stepwise survival analysis.
An explicit assumption of the Four-Minute Rule is that fetuses can be delivered within 1min from the start of the skin incision. Approximately 90% of deliveries during perimortem cesareans took longer than 1min. How does this compare with Cesareans in other circumstances?
In one retrospective study of roughly 900 Cesareans, the 777 women with incision to birth times of less than 10min had a mean birth interval of 6.32±2min while the 138 women with deliveries that took longer than 10min had mean intervals of 13±2.4min (Maayan-Metzger et al., 2010). Similarly, in a study of 145 women undergoing repeat Cesarean section, birth intervals were longer in the presence of adhesions where the mean interval was 15.6min for those with few adhesions while ground system was 19.8min for those with a greater adhesion score (Greenberg et al., 2011). Perhaps the most important study for considering normative urgent Cesarean section times is that of Pearson and MacKenzie (2013). Incision to birth intervals was recorded for 1379 Cesareans stratified by urgency of indication using a previously established classification scheme (Lucas et al., 2000; Bick, 2004). Fifty-five women fell into the most urgent category in which maternal or fetal life was at immediate risk. The median incision to birth interval was 2min with an interquartile range of 2–4min. Finally, in a study of 37,110 cesareans by the Maternal–Fetal Medicine Network, only 3323 had incision to birth intervals of 3min or less (9%) (Alexander et al., 2006).
There are two published studies of perimortem Cesarean section drills by the one group of investigators that suggest that birth within 5min is unlikely at best (Lipman et al., 2010, 2011). In one paper the fastest birth was accomplished by performing the Cesarean in the labor room, but even then only 29% of maternity teams were able to accomplish birth within 5min. In their other study, only 17% of teams were able to deliver in 5min. It is worth noting that there was abundant, dedicated staffing for these drills so their time intervals seem likely to be much better than actual real-world experiences. On the basis of what is known about operating times for cesarean sections both as reported here during maternal cardiac arrest and in other, more common clinical situations, a one-minute incision to birth time is generally an unattainable standard.

Open questions and concluding remarks To answer

Open questions and concluding remarks
To answer those questions and to effectively track the ZIKV epidemic, the main effort should be to develop optimal diagnostic techniques. This will allow tracking all the real ZIKV infection cases for epidemiological studies and correlation analyses with microcephaly/GBS. It will facilitate surveillance and cohort studies, to determine the real percentages of vertically transmitted ZIKV and of fetal death, miscarriage, microcephaly or other fetal malformations. A registry has been created in the US to track and follow up children of mothers infected during pregnancy, which will provide important information regarding the long-term effects of ZIKV infection during pregnancy (Oduyebo et al., 2016).

MicroRNAs (miRNAs) can be isolated from a wide range of different tissues and biological specimens, including biofluids. In 2008, pioneer studies demonstrated that miRNAs were detectable in the tropisetron and serum and that they were particular resistant to degradation by RNAses in the blood (). Subsequent studies reported the presence of cell-free miRNAs (cf-miRNAs) in virtually all other body fluids (), and raised several questions regarding the cf-miRNA stability in the body fluids, the mechanisms of release from cells, and their biological functions.
Cf-miRNAs were found within microvesicles/exosomes, apoptotic bodies (AB), HDL structures, or complexed with AGO proteins (that constitute the miRNA-induced silencing complex, miRISC) () (), which protect them by the action of RNAses. Of note, packaging of miRNAs in exosomes can be controlled by positive selection mechanisms, such as the ceramide-dependent secretory pathway controlled by the nSMase enzyme (e.g. the ceramide biosynthesis neutral sphingomyelinase) (). However, how exactly miRNAs are selected and loaded to exosomes, and how the trafficking is regulated in physiological and pathological conditions, are not yet understood. Another unmet question is about the biological function of cf-miRNAs. In cancer cells, the extracellular release of miRNAs can be a way to reduce the intracellular level of miRNAs with tumor suppressor functions (). On the other hand, cf-miRNAs can function as a paracrine signal to modify tumor microenvironment and support cancer progression; exosomal/MVB/AB-miRNAs may be indeed delivered to neighboring cells where, following uptake, they can modulate the transcription of target-mRNAs () (). An intriguingly alternative mechanism of action for cf-miRNAs was also proposed: the AGO2-complexed miR-21 and miR-29a may act as signaling molecules via binding to intracellular Toll-like receptors (murine TLR-7 and human TLR-8), which are a family of receptors characteristic of immune cells involved in the innate immune system (). The activation of immune cells expressing TLRs stimulates secretion of inflammatory cytokines that ultimately induce tumor cell spread ().
Quantities and species of cf-miRNAs were shown to fluctuate in the presence of malignant and non-malignant disease (). More recently, we and others proposed a reliable method to identify and quantify serum/plasma miRNAs starting from low amounts of serum/plasma (less than 300μl) that could be easily implemented in the clinic for lung cancer early diagnosis. Using this method, two cf-miRNA signatures diagnostic for asymptomatic lung cancer were validated in high-risk individuals (>55years of age, smokers) enrolled in two large Italian lung screening trials (the COSMOS trial, n=1115 and the MILD trial, n=939) (). Despite the fact that these two signatures were derived using different blood components (i.e., serum or plasma; see further below) and from different subjects, they have a substantial fraction of overlapping miRNAs (miR-92a-3p, miR-30c-5p, miR-30b-5p, miR-148a-3p, miR-140-5p), which further confirms the reliability of cf-miRNAs when used as cancer biomarkers. Of note, several other cf-miRNA signatures were recently proposed for the diagnosis of different cancer types (ovary, breast, prostate, liver, colorectal, brain, melanoma, pancreas, etc.). It would be interesting to perform a pan-cancer screening study to understand the ability of these signatures to discriminate among different types of cancer; this will be relevant for the application of these signatures in cancer screening programs.

Our data strengthen and reinforce the relationship

Our data strengthen and reinforce the relationship between renal function and TTR (Kleinow et al., 2011; Kooiman et al., 2014). One study derived from a cohort of anticoagulated AF patients found that patients with moderate and severe CKD had higher TTR than patients with normal renal function, although CKD patients spent more time above therapeutic range and INR-variability significantly increased in patients with moderate and severe CKD (Kooiman et al., 2014). Recently, an analysis from the Outcomes Registry for Better Informed Treatment of Atrial Fibrillation (ORBIT-AF) registry found that an estimated glomerular HG-9-91-01 manufacturer rate below 60ml/min/1.73m2 was significantly associated with a reduced TTR (≤53%) (Pokorney et al., 2015). Our results strongly reinforce this relationship between renal function and anticoagulation control, and that CKD patients derived benefit from VKA treatment with good TTRs (Bonde et al., 2014).
Good quality anticoagulation control (TTR>70%) was an independent predictor for a lower risk of stroke, death and major bleeding. There was a significant inverse correlation between TTR and HR risk function for stroke, death and major bleeding overall, but when this analysis was repeated in the CKD subgroup, the degree of correlation was reduced for stroke and death risk, perhaps reflecting how CKD modulates the risk of adverse events.

Author Contributions

Funding

Disclosures
GYHL: Chairman, Scientific Documents Committee, European Heart Rhythm Association (EHRA). Reviewer for various guidelines and position statements from ESC, EHRA, NICE etc. Steering Committees/trials: Includes steering committees for various Phase II and III studies, Health Economics & Outcomes Research, etc. Investigator in various clinical trials in cardiovascular disease, including those on antithrombotic therapies in atrial fibrillation, acute coronary syndrome, lipids, etc. Consultant for Bayer/Janssen, Astellas, Merck, Sanofi, BMS/Pfizer, Biotronik, Medtronic, Portola, Boehringer Ingelheim, Microlife and Daiichi-Sankyo.
DAL: Investigator-initiated educational grants from Bayer Healthcare and Boehringer Ingelheim. Speaker\’s bureau for Boehringer Ingelheim, Bristol-Myers-Squibb and Bayer for lectures at educational meetings. Dr Lane is also on the Steering Committee of a Phase IV clinical trial sponsored by Bristol-Myers-Squibb.

Introduction
Liver diseases are prevalent diseases in all regions of the world (Lozano et al., 2012), and associated with marked burden of disease (Byass, 2014). Aside from viral hepatitis, the leading cause for liver disease globally, for all stages of liver disease from hepatic steatosis to liver cirrhosis, there is a basic distinction into alcoholic versus non-alcoholic forms based on the history of alcohol intake with various cut-points (mostly between 20 and 40g pure alcohol intake per day [g/day]) (Chalasani et al., 2012; LaBrecque et al., 2014; EASL, 2012; Nascimbeni et al., 2013).
The prevalence of non-alcoholic fatty liver disease (NAFLD), now more common than alcoholic liver disease (Sattar et al., 2014), has doubled in both North America and Asia over the past two decades, and it has become one of the most widespread chronic conditions worldwide (about ¼ of the population), with projected further increase (Byrne and Targher, 2015; Vernon et al., 2011; Younossi et al., 2015). It is frequently associated with impaired glucose tolerance, insulin resistance, hypertension, and obesity (Yki-Jarvinen, 2014), and a risk factor for both type 2 diabetes mellitus and the metabolic syndrome (MetS) (Ballestri et al., 2015), all of which are risk factors for cardiovascular diseases and overall mortality. Hepatic steatosis might also be an independent risk factor for cardiovascular disease (Bonci et al., 2015; Lu et al., 2013; Targher et al., 2010; Loria et al., 2014). Aside from other causes of liver disease (such as use of medication or presence of hereditary disorders known to produce hepatic lipidosis, or viral hepatitis B or C), non-alcoholic hepatic steatosis is diagnosed based on hepatic fatty infiltration in the absence of excessive alcohol consumption (Neuman et al., 2014a). However, there are no systematic investigations as to what excessive alcohol consumption means for hepatic steatosis (Nascimbeni et al., 2013), or whether the relationship between alcohol consumption and hepatic steatosis is continuous or characterized by a threshold effect. Furthermore, the role of sex in this relationship is not clear (Vernon et al., 2011).

Sialic acid residues might also play a role

Sialic order Ketorolac tromethamine salt residues might also play a role in another way. IgG preparations contain large amounts of sialylated antibodies and recent studies have demonstrated that these are crucial for the anti-inflammatory effects of IVIg [reviewed in ()]. Both the F(ab) part as well as the Fc part of the antibodies can contain sialic-acid residues. Yet, while the immunosuppressive role of sialylation of the Fc part is undisputed, data on the role of sialylation of the F(ab) part for anti-inflammatory effects is less clear. Sialylated F(ab′) parts may, however, have yet another function. A recent study indicates that they can serve as decoy receptors for influenza virus thus preventing binding of the virus to its target cells ().

Asthma is a frequent, disabling chronic disease which is characterized by respiratory symptoms (wheeze, breathlessness, chest tightness and cough) and chronic airway inflammation. It affects more than 300 million people world-wide and represents a heavy economic burden to health care systems (). The syndrome asthma is caused by several underlying diseases and trigger factors of which the most frequent ones are IgE-associated allergies and respiratory virus infections (). Although there are obviously a range of different pathomechanisms operative in asthma, for decades the treatment has been based mainly on symptomatic therapy aimed at bronchodilation and reducing inflammation (i.e., pharmacological bronchodilation and corticosteroids). However, the introduction of IgE-targeting therapies for the treatment of allergic asthma has emphasized the need to identify patients by accurate stratification to allow accurate administration of anti-IgE and other new IgE targeting therapies (). Through the characterization of the diseases-causing allergens by molecular cloning techniques, new forms of molecular allergy diagnosis based on allergen molecules have emerged (). order  Ketorolac tromethamine salt The new molecular tests allow not only to discriminate between clinically relevant and irrelevant IgE-sensitizations but also to establish IgE reactivity profiles associated with allergic asthma () and, even to predict the development of respiratory allergy early in childhood (). Furthermore, it seems possible for allergen-derivatives lacking IgE reactivity to discriminate mechanisms of allergen-IgE-mediated allergic inflammation from non-IgE-mediated allergic inflammation, which may help to direct IgE-mast cell-targeting treatments and T cell targeting therapies ().
In patients suffering from allergic asthma, allergen-specific immunotherapy (AIT) is currently emerging as an alternative to symptomatic treatment. In fact, several clinical studies have shown that AIT is effective for asthma (). Moreover, new forms of AIT based on recombinant hypoallergenic allergen-derivatives have been shown to be effective in clinical studies (). These new forms of molecular AIT have major advantages over allergen-extract-based AIT such as reduced side effects and thus increased safety, coverage of the relevant allergens, convenient, few-dose applications as well as lack of allergenicity and thus have great potential to improve the treatment of allergic asthma.
Regarding asthma caused by respiratory virus infections the causal relationship between human rhinovirus HRV, which is thought to be the most relevant virus involved in asthma exacerbation, has been only established by relatively ambiguous nucleic acid-based test methods. However, it has been found that HRV infections induce IgG and IgA responses mainly against an N-terminal peptide of the HRV coat protein VP1. Then it has been demonstrated that serologically detectable increases of the VP1-specific antibody responses occur in patients who have experienced HRV-induced asthma attacks by natural infections as well as by experimental inoculation and can be measured by serological testing (). Available data thus suggest that the increases of VP1-specific antibodies reflect the severity of airway symptoms and may allow identification of the disease-causing HRV groups, indicating that serological tests will become available soon, and allow the identification of persons suffering from HRV-induced asthma attacks.

Torre and colleagues used a planarian experimental infection with

Torre and colleagues used a planarian experimental infection with as a model to study the properties of innate immune memory, with relevance for vertebrate immunity as well. In this model, infection of planarians with changes innate immune responses in an adaptive manner, resulting in an improved rate of pathogen clearance upon subsequent reinfection. Indeed planarians are renowned for their capacity to fight infection and remarkable regenerative abilities. In the pursuit of a mechanistic link between these processes, Torre et al. identified two important novel mechanisms central to the induction of trained immunity (or , as defined by the authors). First, the authors demonstrate the importance of a specific population of pluripotent stem JAK STAT Compound Library called neoblasts for innate immune memory. Second, through a series of experiments using RNA interference, the researchers revealed that genes important for innate immunity confer sustained resistance to via a signaling cascade that is contingent on the Smed-setd8–1 lysine methyltransferase.
These observations are significant for understanding responses during infection and vaccination in humans. One important aspect for which the study of Torre and colleagues is significant is for providing important clues on the physiological mechanisms mediating trained immunity in humans at the level of immune progenitor cells. The long-term protection conferred by vaccination with Bacillus Calmette–Guérin (BCG) far exceeds the lifespan of innate immune cells in the circulation (). The capacity to induce innate immune memory in pluripotent neoblasts in planarians advocates the possibility that innate immune cell precursors in vertebrates can also mount epigenetic and functional reprogramming and thus mediate innate immune memory. Indeed, myeloid cell progenitors have been demonstrated to mediate long-term TLR2-induced tolerance (), and a similar role may be expected for trained immunity.
An important observation is also that Smed-setd8–1 in planarians is homologous to human SET8 (also known as KMT5A), indicating potential for a similar regulatory function in vertebrates. Studies exploring epigenetic changes associated with innate immune memory have focused predominantly on post-translational modifications of H3 histones. Torre et al. now provide the impetus to expand this search to the tails of H4 histones, which are methylated only at lysine 20. Methylation of H4 histones has previously been associated with transcriptional memory in diabetic rodents (), although the precise regulatory function of this modification remains controversial (). Importantly the addition of a single methyl group to H4 histones is associated with transcriptional activation (), and SET8 is the only enzyme known to write this modification ().
To conclude, the elegant study by Torre et al. describes a system of acquired resistance in planarians that shares several important features with trained immunity in vertebrates. Infection with initiates a program of heightened defense against the same pathogen. It remains to be seen how closely this system mirrors the broad non-specific memory of trained immunity. Nevertheless, the central role of neoblasts and Smed-setd8–1 informs about potential new research paths in the search for epigenetic regulators of innate immune memory in vertebrates. Identification of these key factors will greatly accelerate the realization of novel therapeutic approaches to the treatment of infectious and auto-inflammatory diseases, as well as the improvement of vaccination programs ().
Disclosure

Acknowledgements
MGN was supported by an ERC Consolidator Grant (#310372) and a Spinoza grant of the Netherlands Organization for Scientific Research. NPR and MGN received funding from the European Union Horizon 2020 research and innovation program under grant agreement No 667837.

Zika virus (ZIKV) is a mosquito-borne flavivirus that was originally identified in 1947 from a sentinel Rhesus monkey in the Zika forest in Uganda (). Prior to 2007, ZIKV infections occurred periodically in Africa and Asia with mild, self-limiting febrile illnesses such as rash, headache, conjunctivitis, myalgia, and arthralgia. However, in the past decade, ZIKV became explosive in causing outbreaks and epidemics, first on Yap Island in the Federated States of Micronesia in 2007, second in French Polynesia in 2013, third in northeastern Brazil in late 2014, followed by a rapid spread to other countries in the Americas in 2015–2016, including autochthonous transmissions in Florida and Texas in the United States (). During the recent outbreaks and epidemics in Asian and the Americas, ZIKV infection has caused devastating severe diseases, particularly Guillain-Barre syndrome in adults and congenital malformations in fetus, among which congenital malformation is unique to ZIKV infection when compared with diseases caused by other flavivirus infections. Guillain-Barre syndrome is an autoimmune disease characterized by ascending paralysis and polyneuropathy that could occur during the acute or convalescent phases of ZIKV infection (). During pregnancy (especially in the first trimester), ZIKV infections of fetus have been associated with a variety of clinical manifestations, now collectively known as congenital Zika syndrome, including microcephaly, craniofacial disproportion, spasticity, seizures, ocular abnormalities, cerebral calcification, and miscarriage (). Moreover, the disease spectrum of congenital Zika syndrome is expected to grow as some of the infected babies with a normal head circumference may manifest new disease symptoms as they develop; clinical and epidemiological studies are ongoing to uncover the prognosis of these congenitally infected baby patients (). The molecular mechanisms of ZIKV-mediated Guillain-Barre syndrome and congenital malformations remain to be determined. One of the notable driving forces for the congenital Zika syndrome could be the neurotropic nature of ZIKV infection that preferentially targets cortical neural progenitor cells and, to a lesser extent, neuronal cells in other stages of maturity ().

To summarize our work highlights the facts that ECs maintain

To summarize, our work highlights the facts that ECs maintain HIV-specific memory B cell responses associated to effective antiviral humoral activities and that Env-specific memory B cell responses are positively associated with the neutralization breadth in HLA-B*57+ ECs. We propose that promoting HIV-specific B cell polyfunctional responses by therapeutic vaccination might be highly beneficial in cART treated patients.
The following are the supplementary data related to this article.

Funding Sources

Conflicts of Interest

Author Contributions

Acknowledgments

Introduction
The Tat protein of the human immunodeficiency virus type 1 (HIV-1) is a small protein (101aa in most clinical isolates, or 86aa in the widely utilized HXB2 laboratory strain), acting as a transcriptional activator of viral gene expression. At the viral long terminal repeat (LTR) promoter, the protein binds a cis-acting RNA element (trans-activation-responsive region, TAR) present at the 5′-end of each viral transcript (Berkhout et al., 1989). Through this interaction, Tat activates HIV-1 transcription by promoting the assembly of transcriptionally active complexes at the LTR by multiple protein-protein interactions (Giacca, 2004; Ott et al., 2011).
Besides regulating HIV-1 gene expression, >20years ago it was first demonstrated that Tat also possesses the unusual property of entering cells when present in the extracellular milieu (Frankel and Pabo, 1988; Green and Loewenstein, 1988). This property was later extensively characterized and shown to depend on a 9-aa long, arginine-rich sequence (aa 49–57), corresponding to the Tat basic domain, which also mediates nuclear transport and TAR binding. Work performed in different laboratories has shown that short peptides corresponding to this amino sphingosine 1 phosphate receptor stretch can be used as biotechnological tools for the intracellular delivery of heterologous proteins, drugs, viral vectors, siRNAs and nanoparticles (Fittipaldi and Giacca, 2005; Jones and Sayers, 2012; Schmidt et al., 2010; Zhang and Wang, 2012).
We, and others have previously shown that extracellular Tat binds heparin through its basic domain (Mann and Frankel, 1991; Rusnati et al., 1997). We also showed that membrane bound-heparan sulfate proteoglycans (HSPG) are the cell surface receptor for Tat internalization, since cells that are genetically impaired in the synthesis of these molecules fail to internalize the extracellular protein (Tyagi et al., 2001).
A few studies have also provided evidence in support of extracellular Tat release from the expressing cells (Becker-Hapak et al., 2001; Chang et al., 1997; Tasciotti and Giacca, 2005; Tyagi et al., 2001). The mechanism underlying this process, however, has remained largely elusive. The protein does not contain an N-terminal signal peptide driving its secretion from the ER-Golgi pathway and, accordingly, protein export is insensitive to drugs which disrupt the integrity of such organelles (Chang et al., 1997). Thus, Tat is a member of the small group of heterogeneous proteins that exit the cells by a process termed “unconventional” or “non-classical” protein secretion (Nickel and Rabouille, 2009). Recent data show that recruitment of Tat to the inner leaflet of the plasma membrane involves binding to membrane-associated phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) (Rayne et al., 2010), with the consequent formation of membrane pores (Zeitler et al., 2015); similar events occur for the unconventional secretion of FGF-2 (Temmerman et al., 2008). The mechanism for extracellular release of Tat and the molecular identity of the secretory machinery involved, however, remain elusive.

Materials and Methods

Results

Discussion
Our experiments indicate that the α subunit of the cellular Na+,K+-ATPase mediates unconventional Tat secretion in a ouabain-sensitive manner. Extracellular Tat release was affected neither by methylamine (a drug which blocks endosomal recycling and impairs IL1B and FGF-2 non canonical secretion, implying a vesicular intermediate in their release (Hamon et al., 1997; Rubartelli et al., 1990; Zhou et al., 2002)), nor glyburide (a sulfonylurea interfering with the ABC-1 transporter essential for the secretion of Galectin-1 (Flieger et al., 2003; Hamon et al., 1997)). In contrast, Tat secretion was markedly sensitive to ouabain, an inhibitor of the Na+,K+-ATPase. The observations that Tat secretion still occurred in the presence of curcumin, an inhibitor of all P-type ATPases (including the Na+,K+-ATPase) and that the rat D716N α1 mutant, which is impaired in catalytic function (Lane et al., 1993), still rescued Tat secretion in human cells treated with ouabain, are concordant in indicating that the effect of the Na+,K+-ATPase on extracellular Tat release is independent from its enzymatic activity while still demands physical binding of α1 to Tat.

br Acknowledgments br Data Table showed data on the global

Acknowledgments

Data
Table 1 showed data on the global gene expression profile in MCF-7 and MDA-MB-231 cell lines treated with vehicle (DMSO) or DS in vitro. Tables 2–4 showed gene ontology analysis based on molecular functions (Table 2), biological processes (Table 3), and cellular components (Table 4). Various canonical pathways, which were significantly altered between the cell lines (vehicle-treated) or after DS treatment, were presented in Table 5. The genes that were overlapped between these two cell lines (MCF-7 and MDA-MB-231) after DS treatment were listed in Table 6 and in a Venn diagram format in Fig. 1.

Experimental design, materials and methods

Acknowledgments

Data describes human neutrophil activation, measured by isoluminol-enhanced chemiluminescence systems, with two pepducins derived from β2AR (ICL3-8) and CXCR4 (ATI-2341), respectively. Direct neutrophil activation by ICL3-8 and its modulatory effect on FPR signaling are shown (Fig. 1). In addition, data on dose-dependent neutrophil activation induced by ATI-2341 and the effects on this response of FPR specific agonists as well as antagonists are provided (Fig. 2).

Experimental design, material and methods

Acknowledgments
The work was financially supported by the Swedish Research Council (005601 for CD and 02448 for HF), the King Gustaf V Memorial Foundation (FAI2014-0011 for CD and FAI2014-0029 for HF), The Clas Groschinsky Foundation (M1562 for HF), the Swedish State under the ALF agreement (ALFGBG-72510 for CD), the Wilhelm and Martina Lundgren Foundation, and the Ingabritt and Arne Lundberg Research Foundation.

Data
This data mainly focuses on describing the regional marker expression of neural tissues and the data of eye tissue-inducing culture (Figs. 1 and 2), and refers to our recently published [1]. We used mouse melatonin receptor agonist and mouse ESC-derived tissue samples to analyze the regional marker expression via immunostaining and also showed the images of ESC-derived tissues in living condition. The data shown are microscopy images (Fluorescent and Bright-field), graphs (Population of GFP+cells) and schematic diagrams (Step-by-step processes).

Experimental design, materials and methods

Acknowledgments
We are grateful to the members of our laboratory for our many discussions. We also thank J. Levine for his careful English proofreading and G. Oliver for his support of the ESC culture and the EVOS microscope. This work was supported by JSPS KAKENHI Grant Number 26112723 and Research Center Network for Realization of Regenerative Medicine from Japan Agency for Medical Research and development, AMED (to M.E.).

Data
We assessed the storage quality of red blood cells (RBCs) donated by glucose-6-phosphate dehydrogenase (G6PD) deficient, yet eligible, donors compared to control (G6PD sufficient) red blood cells [1]. Intracellular reactive oxygen species (ROS) accumulation was similar in energy depleted (24h/37°C) G6PD− and G6PD+ stored RBCs while stimulation by tert-Butyl hydroperoxide (tBHP) and diamide oxidants resulted in statistically significant increase in ROS accumulation in the G6PD− group (n=6) compared to the G6PD+ group (n=3) (Fig. 1). RBC fragility (both mean corpuscular fragility, MCF and mechanical fragility index, MFI) (Fig. 2) and the characteristics of the microparticles (accumulation, pro-coagulant activity and protein carbonylation index, PCI) (Fig. 3) were equal between the groups under examination throughout the storage period, while only slight differences were observed in the antioxidant capacity of the supernatant (Fig. 4).
Malate levels decreased faster in G6PD− RBCs than in control, G6PD+ RBCs during the storage period in CPD-SAGM (Fig. 5). Finally, N-acetylcysteine (NAC) supplementation (at the concentration used) induced similar changes in both stored RBCs and supernatant (Fig. 6).

Experimental design, materials and methods

Acknowledgments

br Experimental design materials and methods br Funding OR was

Experimental design, materials and methods

Funding
OR was supported by Arthritis Research UK, United Kingdom Grant no. 19,611, and by an EMBO travel award. This work was supported by the Oxford National Institute of Health Research (NIHR) Biomedical Research Center, the Oxford NIHR Biomedical Research Unit (PB).

Acknowledgments

Data
We isolated the porcine peripheral blood and spleen of infected with PRRSV identified by RT-PCR and ELISA, subsets characteristics of DCs were assessed in vivo by flow cytometry (FCM) and fluorescence microscope respectively based on the key surface molecules for pDCs and mDCs. The analyzed data was presented in Fig. 1 and contained two types of data. DCs subtype analysis of porcine peripheral blood (Fig. 1a–b). DCs subtype analysis in porcine spleen (Fig. 1c–d).

Experimental design, materials and methods

Acknowledgments
This study was funded by China Nature Science Project no.31502070 and Dr Start-up fund project of Liaoning no. 20141057. The authors thank all researchers who contributed to the work and we apologize to the researchers whose works could not be discussed here due to space limitations.

Data
One database link, three tables, and one figure are provided in this article. Methyl-seq data from SLE PBMCs segregated based on high or normal numbers of ARID3a+ buy clemastine fumarate was deposited in NCBI׳s GEO database under the following accession number GEO: GSE84965[2]. Tables 1 and 2 show qRT-PCR data obtained via Biomark HD for Type I IFN pathway genes from RNA derived from SLE B cells subdivided based on ARID3a levels [1], and for healthy control B cells with or without CpG induced ARID3a expression [4]. IFN signature genes are in bold. Primers for RT-PCR and qRT-PCR are given in Table 3. Fig. 1 shows the results of RT-PCR of IFNa in four EBV-transformed lymphoblastoid B cell lines [3].

Experimental design, materials and methods

Data
This data article is referred to in the research article entitled Host factors associated with serologic inflammatory markers assessed using multiplex assays[1]. We present the percent differences in distributions of biomarkers of inflammation and immune activation associated with fixed and modifiable host characteristics, minimally-adjusted for age, blood draw time of day, and study site, using serum specimens measured longitudinally (1984–2009) from a sample of HIV-uninfected men in the Multicenter Center AIDS Cohort Study (MACS). Multivariate associations between these biomarkers and sociodemographics and risk behaviors in a sample restricted to 2001–2009 and additionally adjusted for select co-morbidities were also examined.

Experimental design, materials and methods
The MACS has been previously described; briefly, passive transport is a longstanding, prospective cohort study of men who sex with men (MSM) enrolled at four U.S. locations (Baltimore/Washington D.C., Chicago, Los Angeles, and Pittsburgh) to examine the natural and treated histories of HIV-1 infection [2,3]. Study highlights, including data collection forms, may be found at https://statepi.jhsph.edu/macs/macs.html.

Acknowledgements
Samples and data in this manuscript were collected by the Multicenter AIDS Cohort Study (MACS) with support from an American Recovery and Reinvestment Act (ARRA) supplement with centers (Principal Investigators) at: Johns Hopkins University Bloomberg School of Public Health (Joseph Margolick), U01-AI35042; Northwestern University (Steven Wolinsky), U01-AI35039; University of California, Los Angeles (Roger Detels), U01-AI35040; University of Pittsburgh (Charles Rinaldo), U01-AI35041; the Center for Analysis and Management of MACS, Johns Hopkins University Bloomberg School of Public Health (Lisa Jacobson), UM1-AI35043. The MACS is funded primarily by the National Institute of Allergy and Infectious Diseases (NIAID), with additional co-funding from the National Cancer Institute (NCI), United States. This work was also supported by the HIV Prevention Trials Network (HPTN) sponsored by the National Institute of Allergy and Infectious Diseases (NIAID), United States, National Institute on Drug Abuse, United States, National Institute of Mental Health, United States, and Office of AIDS Research, United States, of the NIH, DHHS (UM1 AI068613). Website located at http://www.statepi.jhsph.edu/macs/macs.html. The contents of this publication are solely the responsibility of the authors and do not represent the official views of the National Institutes of Health (NIH).

The recorded actions contain br Experimental design

The recorded actions contain:

Experimental design, materials, and methods
The MMU VAAC database was recorded with the utmost care and attendance by the author. For avoiding effects of any errors and omissions occurred, all recorded scenes were subject to later inspection while doing the preprocessing phase. All actions were acted and recorded in an indoor environment the living room and bedroom of Digital Home Lab at the faculty of engineering in Multimedia University, Malaysia. The sensor used is Microsoft Kinect placed frontal-parallel to the subjects and the child mannequin. It was mounted at the height of 90cm while the distance from the sensor to the subject was 190cm. The effective range of the Kinect sensor is 1.2–3.5m. The sensor motorized pivot can tilt the sensor up to 27° either up or down while has an angular field of view of 57° horizontally and 43° vertically. The horizontal field of the Kinect sensor at the minimum viewing distance of ~0.8m (2.6ft.) is, therefore, ~87cm (34in.), and the vertical field is ~63cm (25in.). Hence, the resulted fluvastatin Supplier is over 1.3mm (0.051in.) per pixel. During the acquisition period, a decision was made to acquire data at the highest resolution and frame rate possible to minimize the effect of noise from the environment or the sensor, so it recorded depth and color frames with 640×480 resolution and speed of 30 frames per second. The child mannequin size is 65cm, and actors׳ size has the range of 150–170cm. The players took the freedom to stand and perform the actions spontaneously. However, they were asked to use both hands and legs. The subjects have been invited to help calibrate the Kinect camera before the start of the video recording. This was done by standing in front of the Kinect camera with arms spread out wide for the joints of the subjects to be detected.
For skeleton joints modality, the following routine was used for creating the data set Fig. 2. We manually segmented all the scenes we have about the eight actions. Moreover, we assigned each sliding window to one action depending on the main activity in each sliding window. Here, we defined and included two more actions (approaching and departing) to the original actions for better and more accurate assigning each window to one main action.

Acknowledgements

Data
The list of all the supplementary data used in this article is summarized in Table 1.

Experimental design, materials and methods
The Estate Surveyors and Valuers Registration Board of Nigeria (ESVARBON) is a body that is statutorily responsible for the regulation of compensations paid by clients to professionals in the Nigerian Institution of Estate Surveyors and Valuers (NIESV). The compensations are in the form of scale upon which the agreed professional fees must be charged. However, the socio-economic realities in Nigeria have necessitated clients to negotiate the charges offered to them by the professionals. It should be noted that mortgage valuation is key to determination of professional fee. Surveys are very vital in understanding and predicting key population characteristics [18–31]. In this case, Ikeja, Lagos, Nigeria was chosen for the research and the study area is indicated in Fig. 1.
The sampling frame is summarized in Table 2.
The sample size is estimated as a percentage of the sample frame using the formula;where:
This research adopted a confidence level of 95%, a sample proportion of 0.05, an allowable error of within 5% of the true prevalence, with the sample frame for registered surveying firms as 82, and the sample frame for commercial banks as 55. The corresponding sample sizes will be calculated using the formula above. The sample size for the registered estate firms is given as:
The sample size for the commercial banks in the study area is calculated as:
Details on the studied population can be accessed in [33,34].