Tissue-specific transcriptional imprinting and heterogeneity in human innate lymphoid cells revealed by full-length single-cell RNA-sequencing

The impact of the microenvironment on innate lymphoid cell (ILC)-mediated immunity in humans remains largely unknown. Here we used full-length Smart-seq2 single-cell RNA-sequencing to unravel tissue-specific transcriptional profiles and heterogeneity of CD127+ ILCs across four human tissues. Correlation analysis identified gene modules characterizing the migratory properties of tonsil and blood ILCs, and signatures of tissue-residency, activation and modified metabolism in colon and lung ILCs. Trajectory analysis revealed potential differentiation pathways from circulating and tissue-resident na\uefve ILCs to a spectrum of mature ILC subsets. In the lung we identified both CRTH2+ and CRTH2− ILC2 with lung-specific signatures, which could be recapitulated by alarmin-exposure of circulating ILC2. Finally, we describe unique TCR-V(D)J-rearrangement patterns of blood ILC1-like cells, revealing a subset of potentially immature ILCs with TCR-δ rearrangement. Our study provides a useful resource for in-depth understanding of ILC-mediated immunity in humans, with implications for disease

Seat testing to investigate the female neck injury risk – preliminary results using a new female dummy prototype

Dynamic performance tests are conducted to assess the neck injury risk. To date seats are assessed based on sled tests mostly using a BioRID which is based on the 50%ile male anthropometry. Since females sustain whiplash associated disorders (WAD) more often than males, their injury risk needs to be addressed as well

Seat testing to investigate the female neck injury risk – preliminary results using a new female dummy prototype

Dynamic performance tests are conducted to assess the neck injury risk. To date seats are assessed based on sled tests mostly using a BioRID which is based on the 50%ile male anthropometry. Since females sustain whiplash associated disorders (WAD) more often than males, their injury risk needs to be addressed as well

Fucosylated Molecules Competitively Interfere with Cholera Toxin Binding to Host Cells

Cholera toxin (CT) enters host intestinal epithelia cells, and its retrograde transport to the cytosol results in the massive loss of fluids and electrolytes associated with severe dehydration. To initiate this intoxication process, the B subunit of CT (CTB) first binds to a cell surface receptor displayed on the apical surface of the intestinal epithelia. While the monosialoganglioside GM1 is widely accepted to be the sole receptor for CT, intestinal epithelial cell lines also utilize fucosylated glycan epitopes on glycoproteins to facilitate cell surface binding and endocytic uptake of the toxin. Further, l-fucose can competively inhibit CTB binding to intestinal epithelia cells. Here, we use competition binding assays with l-fucose analogs to decipher the molecular determinants for l-fucose inhibition of cholera toxin subunit B (CTB) binding. Additionally, we find that mono- and difucosylated oligosaccharides are more potent inhibitors than l-fucose alone, with the LeY tetrasaccharide emerging as the most potent inhibitor of CTB binding to two colonic epithelial cell lines (T84 and Colo205). Finally, a non-natural fucose-containing polymer inhibits CTB binding two orders of magnitude more potently than the LeY glycan when tested against Colo205 cells. This same polymer also inhibits CTB binding to T84 cells and primary human jejunal epithelial cells in a dose-dependent manner. These findings suggest the possibility that polymeric display of fucose might be exploited as a prophylactic or therapeutic approach to block the action of CT toward the human intestinal epithelium

Prognostic value of reading-to-reading blood pressure variability over 24 hours in 8938 subjects from 11 populations.

In previous studies, of which several were underpowered, the relation between cardiovascular outcome and blood pressure (BP) variability was inconsistent. We followed health outcomes in 8938 subjects (mean age: 53.0 years; 46.8% women) randomly recruited from 11 populations. At baseline, we assessed BP variability from the SD and average real variability in 24-hour ambulatory BP recordings. We computed standardized hazard ratios (HRs) while stratifying by cohort and adjusting for 24-hour BP and other risk factors. Over 11.3 years (median), 1242 deaths (487 cardiovascular) occurred, and 1049, 577, 421, and 457 participants experienced a fatal or nonfatal cardiovascular, cardiac, or coronary event or a stroke. Higher diastolic average real variability in 24-hour ambulatory BP recordings predicted (P0.03) total (HR: 1.14) and cardiovascular (HR: 1.21) mortality and all types of fatal combined with nonfatal end points (HR: 1.07) with the exception of cardiac and coronary events (HR: 1.02; P0.58). Higher systolic average real variability in 24-hour ambulatory BP recordings predicted (P0.05) total (HR: 1.11) and cardiovascular (HR: 1.16) mortality and all fatal combined with nonfatal end points (HR: 1.07), with the exception of cardiac and coronary events (HR: 1.03; P0.54). SD predicted only total and cardiovascular mortality. While accounting for the 24-hour BP level, average real variability in 24-hour ambulatory BP recordings added 1% to the prediction of a cardiovascular event. Sensitivity analyses considering ethnicity, sex, age, previous cardiovascular disease, antihypertensive treatment, number of BP readings per recording, or the night:day BP ratio were confirmatory. In conclusion, in a large population cohort, which provided sufficient statistical power, BP variability assesse

Response to Masked hypertension in untreated and treated patients with diabetes mellitus: attractive but questionable interpretations and response to Is masked hypertension related to diabetes mellitus?

We agree with Drs Sobiczewski and Wirtwein that a selfreported diagnosis of diabetes mellitus or the use of antidiabetic drugs may not provide an accurate prevalence of the disease, in particular because of the somewhat different criteria used in the 11 cohorts included in the International Database on Ambulatory blood pressure in relation to Cardiovascular Outcomes (IDACO).1 However, misclassification of diabetes mellitus should weaken, not strengthen, the results of our study. In reply to the comments on ambulatory blood pressure (BP) measurement, we standardized the definitions of daytime and nighttime across cohorts taking into account the differences in the pattern of daily activities between Europe and Asia (see Expanded Methods in the onlineonly Data Supplement).1 Moreover, to account for the varying intervals between ambulatory BP readings, we computed timeweighted BP means of the daytime BP (see Expanded Methods in the online-only Data Supplement).1 In terms of the comparison of cardiovascular risk between untreated diabetic patients with masked hypertension versus untreated diabetic patients with stage-1 hypertension,1 we wish to correct a misconception. The risk was equal with a hazard ratio of 1.07 (95% confidence interval, 0.58\u20131.98; P=0.82). In addition, the hypothesis that we may have underdiagnosed untreated masked hypertension in the patients with diabetes mellitus by using the daytime rather than the nighttime ambulatory BP was put to the test. Of the 229 untreated diabetic patients with normotension by office reading, there were 67 (29.3%) versus 56 (24.5%) patients with masked hypertension as identified by elevated daytime versus nighttime BP, respectively. However, use of daytime versus nighttime BP to define masked hypertension led to discrepant diagnoses in 22.2% of the untreated patients with diabetes mellitus. There is abundant evidence in the diabetic and other high-risk hypertensive states that nocturnal BP, alone or together with daytime BP, is a superior predictor of cardiovascular risk than daytime BP alone.2\u20134 In the current study in untreated patients with diabetes mellitus, masked hypertension, either defined using daytime BP (hazard ratio, 1.96; confidence interval, 0.97\u20133.97; P=0.059) or nighttime BP (hazard ratio, 3.44; confidence interval, 1.68\u20137.06; P<0.001), predicted increased cardiovascular risk as compared with sustained normotension. However, confidence intervals were wide and overlapping probably because of the relatively small number of events in these subgroups. This limitation was noted in our discussion.1 Doumas et al raised several issues. The figure used in the Clinical Implications section,5 which accompanied our publication, 1 addressed many of these issues. First, in both patients with and without diabetes mellitus, there was a significantly greater cardiovascular risk in the untreated masked hypertensive than in the untreated normotensive comparator group. With adjustment for center, sex, and age, the P value was 0.0091. Second, we hypothesized that antihypertensive treatment converted cases of sustained hypertension into treated masked hypertension. This may explain the higher prevalence of masked hypertension in treated versus untreated patients. Similarly, treatment may have converted masked hypertension into sustained normotension. This resulted in a higher rate of cardiovascular events during the 11-year follow-up in both treated masked hypertensives and treated normotensives as compared to the untreated normotensive comparator group with no difference in cardiovascular event rate between treated masked hypetensives and treated normotensives. Third, we postulate that the greatest reduction in cardiovascular risk occurs when antihypertensive treatment dosage is titrated upward or additional antihypertensive agents are added, until the majority of patients with masked hypertension reached sustained normotension, that is, when the ambulatory BP was normalized. Not surprisingly, normalization of ambulatory BP with treatment did not eliminate the lifetime cardiovascular burden associated with previous elevated BP, nor did it correct other cardiometabolic risk factors that cluster with the hypertensive state.6 Thus, the cardiovascular risk in a patient with treated, normalized BP is always greater than in an untreated subject with the identical BP.6 Fourth, antihypertensive treatment increases the prevalence of masked hypertension by decreasing conventional BP versus ambulatory BP by a ratio of 483 to 2.7 This further strengthens our hypothesis that subjects who started with untreated sustained hypertension were converted by treatment into masked hypertension because conventional BP normalized, whereas the ambulatory BP remained elevated. Fifth, because antihypertensive therapy decreases conventional BP more than the ambulatory BP, the reliance on conventional BP as a target treatment goal will result in suboptimal reduction in the cardiovascular event rate. We agree with Doumas et al and so stated in our limitation section: \u201cthe confidence intervals around the hazard ratios comparing the risks in masked hypertensives versus normotensives and stage 1 and stage 2 hypertensives were wide, reflecting limited statistical power to accurately assess differences between these subgroups.\u201d1 Furthermore, the larger nondiabetic IDACO population of patients with masked hypertension was supportive of all the above conclusions and with more robust significant P values.1 Importantly, follow-up BP data and more information on the type of antihypertensive therapy would have been useful, but the absence of this information did not weaken the conclusions of our study.1 Similarly, we agree that 24-hour ambulatory BP cannot be justified on a cost\u2013benefit basis for every normotensive diabetic patient, as has been concluded by other investigators.8 As we suggested in the Perspectives,1 home BP monitoring may be a satisfactory substitute for ambulatory BP monitoring in low-risk patients.9 Furthermore, others have concluded that patients with diabetes mellitus and masked hypertension have sufficient cardiovascular risk factors, frequently including target organ damage, to warrant antihypertensive therapy.10,11 However, we agree with Doumas et al that the benefit of treatment of masked hypertension in patients with and without diabetes mellitus must be confirmed by randomized controlled trials. Finally, as stated in our recent IDACO study,1 our conclusions are hypothesis generating and need verification by further studies

Prognostic value of isolated nocturnal hypertension on ambulatory measurement in 8711 individuals from 10 populations

none26In previous studies, of which several were underpowered, the relation between cardiovascular outcome and blood pressure (BP) variability was inconsistent. We followed health outcomes in 8938 subjects (mean age: 53.0 years; 46.8% women) randomly recruited from 11 populations. At baseline, we assessed BP variability from the SD and average real variability in 24-hour ambulatory BP recordings. We computed standardized hazard ratios (HRs) while stratifying by cohort and adjusting for 24-hour BP and other risk factors. Over 11.3 years (median), 1242 deaths (487 cardiovascular) occurred, and 1049, 577, 421, and 457 participants experienced a fatal or nonfatal cardiovascular, cardiac, or coronary event or a stroke. Higher diastolic average real variability in 24-hour ambulatory BP recordings predicted (P0.03) total (HR: 1.14) and cardiovascular (HR: 1.21) mortality and all types of fatal combined with nonfatal end points (HR: 1.07) with the exception of cardiac and coronary events (HR: 1.02; P0.58). Higher systolic average real variability in 24-hour ambulatory BP recordings predicted (P0.05) total (HR: 1.11) and cardiovascular (HR: 1.16) mortality and all fatal combined with nonfatal end points (HR: 1.07), with the exception of cardiac and coronary events (HR: 1.03; P0.54). SD predicted only total and cardiovascular mortality. While accounting for the 24-hour BP level, average real variability in 24-hour ambulatory BP recordings added 1% to the prediction of a cardiovascular event. Sensitivity analyses considering ethnicity, sex, age, previous cardiovascular disease, antihypertensive treatment, number of BP readings per recording, or the night:day BP ratio were confirmatory. In conclusion, in a large population cohort, which provided sufficient statistical power, BP variability assessedmixedFAN HQ; LI Y; THIJS L; HANSEN TW; BOGGIA J; KIKUYA M; BJORKLUND-BODEGARD K; RICHART T; OHKUBO T; JEPPESEN J; TORP-PEDERSEN C; DOLAN E; KUZNETSOVA T; STOLARZ-SKRZYPEK K; TIKHONOFF V; MALYUTINA S; CASIGLIA E.; NIKITIN Y; LIND L; SANDOYA E; KAWECKA-JASZCZ K; IMAI Y; IBSEN H; O'BRIEN E; WANG JG; STAESSEN JAFan, Hq; Li, Y; Thijs, L; Hansen, Tw; Boggia, J; Kikuya, M; BJORKLUND BODEGARD, K; Richart, T; Ohkubo, T; Jeppesen, J; TORP PEDERSEN, C; Dolan, E; Kuznetsova, T; STOLARZ SKRZYPEK, K; Tikhonoff, Valerie; Malyutina, S; Casiglia, Edoardo; Nikitin, Y; Lind, L; Sandoya, E; KAWECKA JASZCZ, K; Imai, Y; Ibsen, H; O'Brien, E; Wang, Jg; Staessen, J

Risk stratification by 24-hour ambulatory blood pressure and estimated glomerular filtration rate in 5322 subjects from 11 populations.

No previous study addressed whether in the general population estimated glomerular filtration rate (eGFR [Chronic Kidney Disease Epidemiology Collaboration formula]) adds to the prediction of cardiovascular outcome over and beyond ambulatory blood pressure. We recorded health outcomes in 5322 subjects (median age, 51.8 years; 43.1% women) randomly recruited from 11 populations, who had baseline measurements of 24-hour ambulatory blood pressure (ABP(24)) and eGFR. We computed hazard ratios using multivariable-adjusted Cox regression. Median follow-up was 9.3 years. In fully adjusted models, which included both ABP(24) and eGFR, ABP(24) predicted (P 640.008) both total (513 deaths) and cardiovascular (206) mortality; eGFR only predicted cardiovascular mortality (P=0.012). Furthermore, ABP(24) predicted (P 640.0056) fatal combined with nonfatal events as a result of all cardiovascular causes (555 events), cardiac disease (335 events), or stroke (218 events), whereas eGFR only predicted the composite cardiovascular end point and stroke (P 640.035). The interaction terms between ABP(24) and eGFR were all nonsignificant (P 650.082). For cardiovascular mortality, the composite cardiovascular end point, and stroke, ABP(24) added 0.35%, 1.17%, and 1.00% to the risk already explained by cohort, sex, age, body mass index, smoking and drinking, previous cardiovascular disease, diabetes mellitus, and antihypertensive drug treatment. Adding eGFR explained an additional 0.13%, 0.09%, and 0.14%, respectively. Sensitivity analyses stratified for ethnicity, sex, and the presence of hypertension or chronic kidney disease (eGFR <60 mL/min per 1.73 m(2)) were confirmatory. In conclusion, in the general population, eGFR predicts fewer end points than ABP(24). Relative to ABP(24), eGFR is as an additive, not a multiplicative, risk factor and refines risk stratification 2- to 14-fold less

Outcome-driven thresholds for ambulatory pulse pressure in 9938 participants recruited from 11 populations.

Evidence-based thresholds for risk stratification based on pulse pressure (PP) are currently unavailable. To derive outcome-driven thresholds for the 24-hour ambulatory PP, we analyzed 9938 participants randomly recruited from 11 populations (47.3% women). After age stratification (<60 versus 6560 years) and using average risk as reference, we computed multivariable-adjusted hazard ratios (HRs) to assess risk by tenths of the PP distribution or risk associated with stepwise increasing (+1 mm\u2009Hg) PP levels. All adjustments included mean arterial pressure. Among 6028 younger participants (68\u2009853 person-years), the risk of cardiovascular (HR, 1.58; P=0.011) or cardiac (HR, 1.52; P=0.056) events increased only in the top PP tenth (mean, 60.6 mm\u2009Hg). Using stepwise increasing PP levels, the lower boundary of the 95% confidence interval of the successive thresholds did not cross unity. Among 3910 older participants (39\u2009923 person-years), risk increased (P 640.028) in the top PP tenth (mean, 76.1 mm\u2009Hg). HRs were 1.30 and 1.62 for total and cardiovascular mortality, and 1.52, 1.69, and 1.40 for all cardiovascular, cardiac, and cerebrovascular events. The lower boundary of the 95% confidence interval of the HRs associated with stepwise increasing PP levels crossed unity at 64 mm\u2009Hg. While accounting for all covariables, the top tenth of PP contributed less than 0.3% (generalized R(2) statistic) to the overall risk among the elderly. Thus, in randomly recruited people, ambulatory PP does not add to risk stratification below age 60; in the elderly, PP is a weak risk factor with levels below 64 mm\u2009Hg probably being innocuous

Significance of White-Coat Hypertension in Older Persons With Isolated Systolic Hypertension: A Meta-Analysis Using the International Database on Ambulatory Blood Pressure Monitoring in Relation to Cardiovascular Outcomes Population.

The significance of white-coat hypertension in older persons with isolated systolic hypertension remains poorly understood. We analyzed subjects from the population-based 11-country International Database on Ambulatory Blood Pressure Monitoring in Relation to Cardiovascular Outcomes database who had daytime ambulatory blood pressure (BP; ABP) and conventional BP (CBP) measurements. After excluding persons with diastolic hypertension by CBP (90 mm Hg) or by daytime ABP (85 mm Hg), a history of cardiovascular disease, and persons 18 years of age, the present analysis totaled 7295 persons, of whom 1593 had isolated systolic hypertension. During a median follow-up of 10.6 years, there was a total of 655 fatal and nonfatal cardiovascular events. The analyses were stratified by treatment status. In untreated subjects, those with white-coat hypertension (CBP 140/90 mm Hg and ABP 135/85 mm Hg) and subjects with normal BP (CBP 140/90 mm Hg and ABP 135/85 mm Hg) were at similar risk (adjusted hazard rate: 1.17 [95% CI: 0.87\u20131.57]; P0.29). Furthermore, in treated subjects with isolated systolic hypertension, the cardiovascular risk was similar in elevated conventional and normal daytime systolic BP as compared with those with normal conventional and normal daytime BPs (adjusted hazard rate: 1.10 [95% CI: 0.79\u20131.53]; P0.57). However, both treated isolated systolic hypertension subjects with white-coat hypertension (adjusted hazard rate: 2.00; [95% CI: 1.43\u20132.79]; P0.0001) and treated subjects with normal BP (adjusted hazard rate: 1.98 [95% CI: 1.49\u20132.62]; P0.0001) were at higher risk as compared with untreated normotensive subjects. In conclusion, subjects with sustained hypertension who have their ABP normalized on antihypertensive therapy but with residual white-coat effect by CBP measurement have an entity that we have termed, \u201ctreated normalized hypertension.\u201d Therefore, one should be cautious in applying the term \u201cwhite-coat hypertension\u201d to persons receiving antihypertensive treatment